海洛因海绵状白质脑病神经病理及少突胶质细胞凋亡机制研究
|
摘要
海洛因海绵状白质脑病(heroin spongiform leukoencephalopathy HSLE)是指经鼻烫吸海洛因干馏物后出现的以脑白质海绵状空泡变性为病理特征,以小脑性共济失调为主要症状的中枢神经系统器质性疾病。1982年Wolters E在荷兰阿姆斯特丹首次报道47例。2000年陆兵勋教授等首次在我国报道28例HSLE患者。目前世界多个国家和地区均有个案报道。影像学检查发现HSLE呈广泛对称性白质病变,以小脑半球、内囊后肢、胼胝体压部及顶枕叶白质易受损。脑脊液检查碱性髓鞘蛋白(myelin bases protein MBP)增高。其发病可能与中枢神经系统脱髓鞘有关。单独静脉注射海洛因者未发现HSLE发病。从零售的海洛因粉剂中已鉴定出大量搀杂物,如安定、异烟肼、滑石粉等物质,推测可能是某种搀杂物引起HSLE,而不是海洛因本身。但病因和发病机制不清。有研究证实多种因素均可引起脑白质脱髓鞘和少突胶质细胞凋亡,最终导致轴突脱髓鞘及传导性能丧失。调节细胞凋亡因子bcl-2/bax比值减少可能直接导致神经元凋亡通路。环己铜草酰二腙可引起脑白质脱髓鞘改变,并具有明显的区域分布,胼胝体压部及后穹隆病灶明显,而胼胝体膝部及前小脑脚则没有明显的脱髓鞘改变。本研究以影像学及神经病理学分析HSLE脑白质脱髓鞘及脑水肿特征,用海洛因搀杂物及环己铜草酰二腙探索制备大鼠脑白质改变模型,并与HSLE相比较,从少突胶质细胞凋亡及调控因子bcl-2/bax着手,研究HSLE脑水肿特征及少突胶质细胞凋亡的发病机制。
1海洛因海绵状白质脑病影像学及神经病理特征研究
1.1研究目的
通过MRI检查、HE染色和MBP免疫组化方法,研究和探讨HSLE脱髓鞘和海绵状空泡变性特征。
1.2研究方法
HSLE住院患者6例(南方医科大学南方医院),头颅MRI采集T1WI、T2WI、水抑制、增强及MRA影像,综合分析脑部病变定位、定性及脑血管改变情况。
4例HSLE尸检脑组织,5例正常对照脑组织(来源于非脑部病变死亡尸检病例)。HE染色、MBP免疫组化及透射电镜检测脑白质脱髓鞘、空泡变性及超微结构特征。
1.3研究结果
1.3.1头颅MRI检查
大脑皮质变薄,脑白质扩大,病灶周边无明显脑水肿,无明显占位效应。6例患者均存在小脑白质、胼胝体压部、内囊后支及枕叶白质病灶。病灶呈长T1WI、长T2WI。双侧大脑FLAIR像病灶呈高信号,较T2WI稍减低,增强无强化。MRA显示各主干动脉纤细,分支减少,与临床症状严重程度成正比。
1.3.2透射电镜
HSLE脑白质脱髓鞘改变,少突胶质细胞多发性空泡变性,在其残留的胞浆中可见肿胀的线粒体及膨胀的内质网。
1.3.3 HE染色
HSLE患者脑白质疏松,大脑白质、小脑白质、胼胝体空泡形成,沿轴索走行。大脑白质由浅层到深层,空泡样改变逐渐明显,结构散乱。小脑颗粒下层白质空泡较大,向深层白质空泡渐小而密集。白质血管周围空泡形成明显。皮质下白质、胼胝体轴索肿胀,细胞核分散。
1.3.4 MBP染色
髓鞘染色成棕褐色,大脑白质、小脑白质及胼胝体染色浅,染色不均,空泡样无着色区由浅层到深层逐渐增多、加重,髓鞘纤维断裂、崩解,白质疏松。
1.4研究结论
(1)HSLE白质病变区影像学髓鞘明显减少;
(2)HSLE形态学脑白质脱髓鞘和空泡变;
(3)HSLE存在血管痉挛及继发性微循环改变;
(4)HSLE白质空泡变性与血管改变有关。
2 bcl-2/bax基因表达和少突胶质细胞凋亡在海洛因海绵状白质脑病发病机制中的作用研究
2.1研究目的
检测HSLE脑组织细胞凋亡及其调节因子,研究HSLE少突胶质细胞凋亡的发病机制
2.2研究方法
4例HSLE尸检脑组织,5例正常对照脑组织(来源于非脑部病变死亡尸检病例)。检测大脑白质、小脑白质和胼胝体区TUNEl阳性细胞、caspase-3阳性表达及bcl-2/bax基因表达并计数,统计分析采用SPSS 13.0软件包,各阳性细胞数据用均数±标准差(x~-±s)表示,病例组与对照组不同脑区比较采用重复测量数据的方差分析,多重比较采用LSD方法,相同脑区两组间比较采用独立样本t检验,TUNEL细胞凋亡计数与caspase-3表达之间的关系采用-Pearson相关检验。
2.3研究结果
2.3.1 TUNEL细胞凋亡检测
HSLE患者大脑白质有少量凋亡阳性细胞,小脑白质、胼胝体部位大量细胞核呈棕褐色,深染不均,计数明显高于对照组,与对照组相比差异有显著统计学意义(F=327.913,P=0.000);部位间比较差异有统计学意义(F=68.100,P=0.000),部位间多重比较,小脑白质与胼胝体之间差异无统计学意义(P=0.310),大脑白质与小脑、胼胝体比较差异有统计学意义(P=0.018,P=0.017);两组不同脑区之间交互效应具有统计学意义(F=83.715,P=0.000)。
2.3.2 Caspase-3免疫组化阳性表达
caspase-3阳性细胞广泛存在于HSLE大脑、小脑和胼胝体白质,以病灶中心部位为最明显,在边缘区及相对病变程度轻的额叶,也有少量细胞凋亡。两组间比较差异有统计学意义(F=387.955,P=0.000);部位间比较差异有显著统计学意义(F=215.727,P=0.000),部位间多重比较,小脑白质与胼胝体之间差异无统计学意义(P=0.096),大脑白质与小脑白质(P=0.020)、胼胝体(P=0.022)比较差异有显著统计学意义;TUNEL阳性计数与caspase-3阳性计数之间存在正相关(pearson r=0.924,P=0.000)。脑白质主要由少突胶质细胞形成髓鞘,表明HSLE脑白质存在广泛大量少突胶质细胞凋亡。
2.3.3 Bcl-2基因表达
HSLE与对照组间比较差异无统计学意义(F=0.001,P=0.978),大脑白质、小脑白质及胼胝体部位间比较差异无统计学意义(F=0.001,P=0.999)。
2.3.4 Bax基因表达
两组间比较差异有显著统计学意义(F=11.098,P=0.013),HSLE部位间比较差异有显著统计学意义(F=5.632,P=0.016),部位间多重比较,小脑白质与胼胝体之间差异无统计学意义(P=0.952),大脑白质与小脑白质(P=0.032)、胼胝体(P=0.035)比较差异有显著统计学意义。Bcl-2为促生存家族,bax为促凋亡家族,HSLE脑白质bax表达增高与细胞凋亡有关,Bax高表达使线粒体细胞色素C释放,致使Caspases-9和Caspases-3激活而诱导凋亡,Bax与Bcl-2的相对比值在决定细胞生存或死亡中可能起关键作用。
2.4研究结论
(1)少突胶质细胞凋亡是脱髓鞘的原因;
(2)TNF受体1结合后激活caspase-3导致少突胶质细胞凋亡;
(3)Bcl-2/Bax比值减少促使少突胶质细胞凋亡。
3环己铜二腙诱导大鼠脑白质空泡样变性特征及少突胶质细胞凋亡研究
3.1研究目的
通过探索并建立脑白质海绵状空泡变性模型,进一步研究其病因及发病机制,为HSLE病因及发病机制研究提供新的研究内容和思路。
3.2研究方法
SD大鼠72只,动物分组:搀杂物烟熏组(搀熏组)、搀杂物喂食组(搀喂组)、环己铜二腙组(铜腙组)、生理盐水组(对照组),每组分2周、4周、6周3个时间段,每时间段6只SD大鼠。搀杂物按安定:苯巴比妥:异烟肼:水合硅酸镁=2.5∶30∶100∶200的比例研磨成粉备用。搀杂物混合粉剂染毒量每次25mg/100g体重。铜腙组将环己铜二腙配制成1%混悬液,按10mg/100g体重经口灌胃;对照组予等量生理盐水灌胃。均为1次/日。制备SD大鼠脑白质空泡变性膜型,在相应时间点脑组织取材、固定。
HE染色、MBP免疫组化髓鞘染色检测脑白质脱髓鞘及空泡变性特征;透射电镜观察胼胝体压部超微结构;TUNEL技术及caspase-3免疫组化检测细胞凋亡。
统计学方法:应用SPSS 13.0软件包,TUNEL及caspase-3阳性细胞数据用均数±标准差(x~-±s)表示,各组相同脑区及不同时间点之间进行析因方差分析。
3.3研究结果
3.3.1实验鼠大体脑标本
72例SD大鼠脑表面灰白色,无充血、肿胀,各组大体观察无明显差异。
3.3.2 HE染色
搀熏组、搀喂组、铜腙2、4周组及对照组大、小脑结构正常,白质内未见空泡样改变。铜腙6周组可见大鼠脑白质空泡形成,空泡呈长条形或椭圆形,沿纤维走行,以脑干和枕顶叶白质空泡样改变最明显,血管无明显改变,血管周围无明显空泡,无明显炎细胞浸润,未见坏死区,未见明显胶质细胞增生,大脑灰质神经细胞和小脑灰质Purkinje细胞未见缺失和变性。
3.3.3透射电镜
铜腙4周组大鼠,可见髓鞘排列较紊乱,部分结构松解变性,但无典型脱髓鞘改变,铜腙6周组大鼠脑胼胝体可见髓鞘肿胀,髓鞘板层内有空泡形成。
3.3.4 MBP免疫组化染色
搀熏组、搀喂组、铜腙2、4周组及对照组大鼠脑白质、胼胝体染色均匀,未见组织疏松、空泡形成等改变。铜腙6周组可见大鼠脑白质淡染、稀疏,髓鞘中断或被挤向两侧,出现无着色空泡,以脑干和顶枕叶白质最明显。
3.3.5 TUNEL染色
铜腙2周组脑白质偶见散在细胞凋亡,铜腙4周组可见脑白质较多凋亡阳性细胞,深染不均,铜腙6周组可见脑室周白质、小脑白质、胼胝体及脑干部位大量凋亡细胞,分布在空泡区,各组TUNEL计数比较差异具有显著统计学意义(F=309.934,P=0.000),不同时间之间相比差异具有显著统计学意义(F=163.839,P=0.000),各组不同时间交互效应有显著统计学意义(F=155.270,P=0.000),铜腙组与搀熏组、搀喂组及对照组比较差异均有统计学意义(P=0.000),6周组与2周组、4周组比较均有显著统计学意义(P=0.000),4周组与2周组比较具有显著统计学意义(P=0.000),以铜腙6周组最高,其次为铜腙4周组,其余各组及各时间之间差异无统计学意义。提示铜腙4周组脑白质已出现细胞凋亡,随着时间延长,到6周时细胞凋亡加重。
3.3.6 caspase-3免疫组化
铜腙4周组可见脑白质较多凋亡阳性颗粒,深染不均,铜腙6周组可见皮质下白质、小脑白质、胼胝体及脑干部位大量凋亡细胞,分布在脑白质空泡区,各组之间差异具有显著统计学意义(F=84.018,P=0.000),不同时间之间差异具有显著统计学意义(F=23.611,P=0.000),铜腙组与搀熏组、搀喂组及对照组比较差异均有统计学意义(P=0.000),6周组与2周组、4周组比较均有显著统计学意义(P=0.000),4周组与2周组比较有显著统计学意义(P=0.003),以铜腙6周组最高,其次为铜腙4周组,其余各组及各时间之间差异无统计学意义。提示铜腙4周组脑白质出现细胞凋亡信号,随着时间延长,到6周时细胞凋亡加重。与TUNEL凋亡染色相一致,与HE染色及MBP髓鞘染色形态学损害区相一致。说明细胞凋亡与脑白质海绵状空泡变性有关,细胞凋亡可能为脑白质海绵状空泡变性的原因之一。
3.4研究结论
(1)安定、苯巴比妥、异烟肼、水合硅酸镁混合物烟熏或喂食未致鼠脱髓鞘和空泡变;
(2)海洛因搀杂物未引起SD鼠白质细胞凋亡;
(3)鼠经口摄入铜腙主要损害靶点在CNS少突胶质细胞,其凋亡致空泡样变;
(4)铜腙喂食致SD鼠脑白质脱髓鞘及空泡变性,无血管相关性;
(5)出现细胞凋亡(4W)早于出现脑白质脱髓鞘和空泡变性(6W),细胞凋亡是脱髓鞘及空泡变的先导;
(6)细胞凋亡与脱髓鞘、空泡形成有关,呈时间剂量相关性,随铜腙摄入时间延长、总剂量增加,细胞凋亡增多。
4全文总结
(1)HSLE白质病变区影像学髓鞘明显减少;
(2)HSLE形态学脑白质脱髓鞘和空泡变;
(3)HSLE存在血管痉挛及继发性微循环改变;
(4)HSLE白质空泡变性与血管改变有关;
(5)少突胶质细胞凋亡是脱髓鞘的原因;
(6)TNF受体1结合后激活caspase-3导致少突胶质细胞凋亡;
(7)Bcl-2/Bax比值减少促使少突胶质细胞凋亡;
(8)安定、苯巴比妥、异烟肼、水合硅酸镁混合物烟熏或喂食未致鼠脱髓鞘和空泡变;
(9)海洛因搀杂物未引起SD鼠白质细胞凋亡;
(10)鼠经口摄入铜腙主要损害靶点在CNS少突胶质细胞,其凋亡致空泡样变;
(11)铜腙喂食致SD鼠脑白质脱髓鞘及空泡变性,无血管相关性;
(12)出现细胞凋亡(4W)早于出现脑白质脱髓鞘和空泡变性(6W),细胞凋亡是脱髓鞘及空泡变的先导;
(13)细胞凋亡与脱髓鞘、空泡形成有关,呈时间剂量相关性,随铜腙摄入时间延长、总剂量增加,细胞凋亡增多。
Heroin spongiform leukoencephalopathy(HSLE)is a kind of central nervous system organic diseases with white matter spongiform vacuolar degeneration for the pathological features after snorting heroin.Cerebellar ataxia is the main symptoms. This disease was first reported by Wolters E in 1982 after he studied 47 patients in Amsterdam Netherlands.Within the following two decades,many individual cases were reported in some country.In 2000,Dr Lu Bing-Xun firstly diagnosed and reported 28 cases HSLE patients in Guangdong province of China.This was the first outbreak in Chinese mainland and patient had a history of inhaling heroin vapor with noses.Medical image showed that cerebral and cerebellar white matter had extensive and symmetric changes,which was spongiform vacuoles degeneration of white matte, myelin bases protein(MBP)was increase in the cerebrospinal fluid.Its incidence may be related to the central nervous system demyelination.But specific mechanisms unclear.Single intravenous injection of heroin were not found HSLE incidence. Substantial adulterant has been identified from the retail heroin powder.Such as diazepam,isoniazid,talc powder,etc.HSLE are probably caused by heroin adulterant, rather than the heroin itself.Research has shown that secondary oligodendrocyte apoptosis can be caused by radiation,hypoxia-ischemia.Bcl-2/bax change can direct cause neuronal apoptosis pathway.Cuprizone can cause demyelination of white matter,and significantly distribute the splenium of corpus callosum and post-fornix, but the genu of corpus callosum and rostral cerebellar pebuncle is no obvious demyelination.This study adopt imaging and neuropathological to analyze features of cerebral edema and demyelination of white matter in HSLE,used heroin adulterant and cuprizone to prepare rat model of white matter changes,and compared to HSLE, detect oligodendrocytes apoptosis and bcl-2/bax to study the HSLE possible originate from blood vessel and oligodendrocyte apoptosis pathogenesis.
1.Study of characteristics of Neural Pathology in Heroin spongiform leucoencephalopathy imaging and neuropathological
1.1 Purposes
Through the MRI examination,HE staining and MBP immunohistochemical method,to study and explore HSLE features of vacuolar degeneration and the demyelination.
1.2 Methods
Six cases of patients HSLE(Nanfang Hospital,Southern Medical University). The incidences were after withdrawal.MRI was used to Collect T1WI,T2WI, enhancement,FLAIR,DWI and MRA image,comprehensive analysis localizion, qualitation and cerebral vessels change.
4 cases of brain autopsies samples of HSLE and 5 controllers.The frontal lobes, cerebella and corpus callosum were drawn.Demyelination and vacuolation were detected by HE stained and MBP immunohistochemisty under the light microscope. Ultra-structrue were detected by electron microscope method.
1.3 Results
1.3.1 MRI of HSLE patients
Cerebral cortex of HSLE patients become thinning,white matter expansion,no significant edema surrounding lesions,no significant mass effect.Lesions appear long T1WI,long T2WI.There are cerebellar white matter,splenium of corpus callosum, internal capsule posterior branch and occipital white matter lesions in 6 patients. FLAIR showed high signal lesions,like than T2WI slightly reduced,suggest myelination decrease.Enhanced no enhancement,refer to the blood-brain barrier without damage.MRA showed internal carotid artery,anterior cerebral artery,middle cerebral artery,posterior cerebral artery running normal,the arterial trunk slender, some rigid,branch red,and in direct proportion to the severity of clinical symptoms.
1.3.2 Electron microscop
HSLE Oligodendrocytes show multiple vacuolar degenerate.Swollen mitochondria and endoplasmic reticulum can be seen in their reliquus cytoplasm.
1.3.3 HE staining
The encephalon white matter of HSLE presented obvious rarefaction and unequal size vacuole changes.Vacuoles run along the axon.The vacuole changed more from superficial layer to deep layer.The structures were heterocentric.Lower white matter of cerebellar granule show larger vacuoles,to the deeper white matter vacuoles gradually small and intensive.Perivascular Vacuolating were more obvious. There is a small amount of vacuolated changes surrounding the small blood vessels and the capillary.
1.3.4 MBP immunohistochemisty
Compare with the control group,cerebral White matter,cerebellar white matter and corpus callosum showing stained slight and uneven,vacuolated areas no staining, There were growing heavier myelin fiber collapse and fracture from superficial layer of white matter to deep layer.HSLE cerebral cortex can be seen a small amount of filament-like fibers,dyeing uniformity,compared with the control group no significant difference.
1.4 Conclusions
(1)Myelination were obvious decreased in HSLE white matter vacuolar area;
(2)HSLE white matter exist Extensive demyelination and vacuolar degeneration;
(3)Secondary vascular changes increase the vacuolar degeneration.We conjecture cerebral microvascular changes related to vacuole formation.
2 Research of bcl-2/bax gene expression and oligodendrocyte apoptosis in the heroin spongiform leukoencephalopathy pathogenesis
2.1 Purposes
Detect cells apoptosis and its regulatory factors in HSLE brain tissue,research the role of oligodendrocyte apoptosis in HSLE pathogenesis.
2.2 Methods
4 cases of brain autopsies samples of HSLE and 5 controllers.The frontal lobes, cerebella and corpus callosum were drawn.Caspase-3 and bcl-2/bax were stained with immunohistochemical method.TUNEL cells apoptosis were detected under the light microscope.Quantitative analysis was conducted by micro-image analysis instrument.Statistical analysis use SPSS 13.0 software package.Data of the positive cells write as(x~-±s).Comparison of different brain regions in HSLE and control group used repeated measure analysis of variance,multiple comparisons using LSD method,the same brain area comparison between the two groups using independent sample t test,TUNEL apoptotic cell counts and expression of caspase-3 used the Pearson correlation test.
2.3 Results
2.3.1 TUNEL positive counts
There were oligodendrocyte apoptosis in white matter HSLE and apoptotic bodies obviously in vacuolar area,which appeared coloration and partial condense of caryon.Count was significantly higher than control group,compared with the control group the difference was statistical significance(F=327.913,P=0.000);site to compare statistically significant differences(F=68.100,P=0.000),multiple comparisons between sites,cerebellar white matter and corpus callosum was no significant difference(P=0.310),cerebral gray matter and cerebellar white matter,the corpus callosum have statistically significant difference(P=0.018,P=0.017). Different brain regions between two groups there were significant interaction effect (F=83.715,P=0.000).
2.3.2 Caspase-3 immunohistochemisty positive expression
There were leonine Caspase-3 protein positive oligodendrocyes in the focal zone of HSLE,Counts was significantly higher than control group,compared with the control group the difference was statistical significance(F=387.955,P=0.000); Comparison at sites statistically significant differences(F=215.727,P=0.000), multiple comparisons between sites,cerebellar white matter and corpus callosum was no significant difference(P=0.096),cerebral gray matter and cerebellar white matter, the corpus callosum have statistically significant difference(P=0.020,P=0.022). There were direct correlation between TUNEL-positive count and caspase-3 positive counts of HSLE(pearson correlation coefficient r=0.924,P=0.000).Myelins were mainly formed by oligodendrocyte.This indicate HSLE exist extensive white matter oligodendrocyte apoptosis.
2.3.3 Bcl-2 gene expression
The difference was not significant between HSLE and the control group (F=0.001,P=0.978),brain gray matter,white matter and corpus callosum to compare the difference was not significant(F=0.003,P=0.999).
2.3.4 Bax gene expression
The differences of HSLE comparison with the control group were statistical significant(F=11.098,P=0.013).Inter-site difference was significant statistical significance(F=5.632,P=0.016).Multiple sites comparison,cerebellar white matter and corpus callosum was no significant difference(P=0.952),cerebral gray matter and cerebellar white matter(P=0.032)and corpus callosum(P=0.035)difference was significant statistical significance.Bcl-2 is promote the survival family.Bax is the pro-apoptotic family.Bax high expression increase the mitochondrial cytochrome C release,resulting in Caspases-9 and Caspases-3 activation and induced cells apoptosis. Bcl-2 and Bax ratio may play a key role in determining the cell survival or death.
2.4 Conclusions
(1)A large number of apoptotic cells in white matter vacuolation areas,speculated oligodendrocyte apoptosis are probably one of HSLE pathogenesis;
(2)Bcl-2/bax expression change are one of oligodendrocyte apoptosis path;
(3)These were related to Oligodendrocyte poisoning and mitochondrial dysfunction.
3 Characteristics of vacuolar degeneration and oligodendrocyte apoptosis in alba of SD rats with cuprizone induced
3.1 Purposes
Through explore and the establish alba vacuolar degeneration model,further study its etiology and pathogenesis,for etiology and pathogenesis of HSLE study provides new research content and ideas.
3.2 Methods
72 Sprague-Dawley(SD)rats.Rats were randomly divided into four groups: adulterant smoking group(AS),adulterant feeding group(AF),cuprizone feeding group(Cu),and control saline feeding group(con).Each group set 3 time point:2,4 and 6 weeks.We adopt heroin adulterant(INH,diazepam,barbitone and talcum powder)and cuprizone induce SD rats leukoencephalopathy model.Adulterant by ratio:diazepam:isoniazid:barbitone:talcum powder=2.5:30:100:200 mix grinding into powder.Adulterant mixed powder exposed to the volume of each rat by weight 25mg/100g respectively.1%cuprizone were fed rats(10mg/100g weight).While the control group was fed with equal saline.Once a day.
The rats' brains were detected with HE staining and MBP immunohistochemisty staining.Cells apoptosis was counted by TUNEL and caspase-3 immunohistochemisty staining under light microscope,ultra-structure was observed by electron microcope.
Statistical method:Apply SPSS 13.0 software package.The positive counts data of TUNEL and caspase-3 of SD rat model as(x~-±s).Comparision between the same brain regions in each group and of different time points used factorial variance analysis respectively.
3.3 Results
3.3.1 The whole brain samples of SD rats
72 cases of SD rat brain were not seen congestive and swelling.Rats of Cuprizone 6 weeks has mild adhesions between the meninges and the skull.Cerebral, cerebellar gray whiter matter structure clear,no expansion of white matter.
3.3.2 HE staining
Brain gray matter and white matter no vacuolation in AS group,AF groups,Cu 2w,4w group and Con group.Rats of Cu 6w group show the white matter vacuolization,vacuoles were bar shape and running along the fibers.The brain stem and occipital lobe white matter vacuolation were the most obvious,no significant changes in blood vessels,no obvious inflammatory cell infiltration,no necrosis,no obvious glial cell hyperplasia,cerebral gray matter nerve cells were not loss and degeneration.
3.3.3 Electron microscop
Cu 4w group of rats show more disordered arrangement of myelin,but no typical demyelination.Cu 6w group rats corpus callosum can be seen myelin swelling and myelin lamellar Vacuolating.
3.3.4 MBP immunohistochemical
White matter was sparse in Cu 6w group rats.Myelin lamellar split, vacuolization without coloring.The brain stem and top-occipital white matter most obvious.
3.3.5 TUNEL positive cells
More positive apoptotic nuclei were seen in white matter of Cu 4w group,deeply stained unevenly.Large number of apoptotic cells were seen in Cu 6w group,which main located in vacuole area of the ventricle white matter,cerebellar white matter, corpus callosum and brainstem.TUNEL counts in each group the difference was significant statistical significance(F=309.934,P=0.000).The difference between different times was significant statistical significance(F=163.839,P=0.000),Cu 6w group and the 2w,4w group were significantly statistical significance(P=0.000),4w group compared with the 2w group had significant statistical significance(P=0.000), with Cu 6w group were the highest,followed by Cu 4w group.prompted Cu 4w groups have emerged apoptosis in white matter,extended over time,to 6w the apoptosis increase.
3.3.6 Caspase-3 positive expression
This result and TUNEL were accordant.Rats brain gray matter in each group there is a small amount of caspase-3 positive expression,the difference was not significant(F=0.485,P=0.694).Cu 4w group revealed higher apoptosis positive granules.large number of apoptotic cells were count in Cu 6w group.The difference of Caspase-3 expression in each group was significant statistical significance (F=84.018,P=0.000).The differences between different time have significant statistical significance(F=23.611,P=0.000),Cu 6w group and 2w group,4w group were significantly statistical significance(P=0.000),4w group compared with 2w group have significant statistical significance(P=0.003),while Cu 6w group were the highest,followed by Cu 4w group.Prompt Cu 4w group white matter appeared apoptotic signal,apoptosis increaseas by 6w.Cells apoptosis is consistent with myelin lesion morphology of HE staining and MBP staining,and it is earlier than morphology change.Description apoptosis cause white matter vacuolar degeneration. Cell apoptosis may be the originating causes of vacuolar degeneration in white matter.
4 Conclusions
(1)Smoking or feeding rats can not induce SD rats alba white matter changes by diazepam,isoniazid,barbitone and talcum powder adulterant;
(2)Cuprizone oral perfusion may cause SD rats alba vacuolar degeneration;
(3)There were a large number of apoptosis in alba lesions of SD rats by cuprizone induced.Suggesting that apoptosis was related to alba demyelination and vacuolar degeneration;
(4)Damage target of cuprizone were in the central nervous system oligodendrocytes. The oligodendrocyte apoptosis was earlier than alba demyelination at the happened time.Guess apoptosis cause demyelination and vacuolar degeneration in white matter of SD rats by cuprizone induced;
(5)Vacuolation was induced by cuprizone in SD rats.Vacuolating time variability were dose-related,with the time prolonged,the total dose increase,demyelination aggravate;
(6)Cuprizone was used to induced white matter degeneration,no significant brain edema,no obvious vascular relevance;
(7)The cuprizone-induced white matter degeneration were different with HSLE incidence reasons,Neuropathological changes were different,but there are cell apoptosis.
Summary of The full text
1、Myelination were obvious decreased in HSLE white matter vacuolar area;
2、HSLE white matter exist Extensive demyelination and vacuolar degeneration;
3、Secondary vascular changes increase the vacuolar degeneration.We conjecture cerebral microvascular changes related to vacuole formation.
4、A large number of apoptotic cells in white matter vacuolation areas,speculated oligodendrocyte apoptosis are probably one of HSLE pathogenesis;
5、Bcl-2/bax expression change are one of oligodendrocyte apoptosis path;
6、These were related to Oligodendrocyte poisoning and mitochondrial dysfunction;
7、Smoking or feeding rats can not induce SD rats alba white matter changes by diazepam,isoniazid,barbitone and talcum powder adulterant;
8、Cuprizone oral perfusion may cause SD rats alba vacuolar degeneration;
9、There were a large number of apoptosis in alba lesions of SD rats by cuprizone induced.Suggesting that apoptosis was related to alba demyelination and vacuolar degeneration;
10、Damage target of cuprizone were in the central nervous system oligodendrocytes.The oligodendrocyte apoptosis was earlier than alba demyelination at the happened time.Guess apoptosis cause demyelination and vacuolar degeneration in white matter of SD rats with cuprizone induced;
11、Vacuolation were induced by cuprizone in SD rats.Vacuolating time variability were dose-related,with the time prolonged,the total dose increase, demyelination aggravate;
12、Cuprizone were used to induced white matter degeneration,slight cells swelling,no obvious vascular relevance;
13、The cuprizone-induced white matter degeneration were different with HSLE incidence reasons,Neuropathological changes were different,but there are cell apoptosis.
1海洛因海绵状白质脑病影像学及神经病理特征研究
1.1研究目的
通过MRI检查、HE染色和MBP免疫组化方法,研究和探讨HSLE脱髓鞘和海绵状空泡变性特征。
1.2研究方法
HSLE住院患者6例(南方医科大学南方医院),头颅MRI采集T1WI、T2WI、水抑制、增强及MRA影像,综合分析脑部病变定位、定性及脑血管改变情况。
4例HSLE尸检脑组织,5例正常对照脑组织(来源于非脑部病变死亡尸检病例)。HE染色、MBP免疫组化及透射电镜检测脑白质脱髓鞘、空泡变性及超微结构特征。
1.3研究结果
1.3.1头颅MRI检查
大脑皮质变薄,脑白质扩大,病灶周边无明显脑水肿,无明显占位效应。6例患者均存在小脑白质、胼胝体压部、内囊后支及枕叶白质病灶。病灶呈长T1WI、长T2WI。双侧大脑FLAIR像病灶呈高信号,较T2WI稍减低,增强无强化。MRA显示各主干动脉纤细,分支减少,与临床症状严重程度成正比。
1.3.2透射电镜
HSLE脑白质脱髓鞘改变,少突胶质细胞多发性空泡变性,在其残留的胞浆中可见肿胀的线粒体及膨胀的内质网。
1.3.3 HE染色
HSLE患者脑白质疏松,大脑白质、小脑白质、胼胝体空泡形成,沿轴索走行。大脑白质由浅层到深层,空泡样改变逐渐明显,结构散乱。小脑颗粒下层白质空泡较大,向深层白质空泡渐小而密集。白质血管周围空泡形成明显。皮质下白质、胼胝体轴索肿胀,细胞核分散。
1.3.4 MBP染色
髓鞘染色成棕褐色,大脑白质、小脑白质及胼胝体染色浅,染色不均,空泡样无着色区由浅层到深层逐渐增多、加重,髓鞘纤维断裂、崩解,白质疏松。
1.4研究结论
(1)HSLE白质病变区影像学髓鞘明显减少;
(2)HSLE形态学脑白质脱髓鞘和空泡变;
(3)HSLE存在血管痉挛及继发性微循环改变;
(4)HSLE白质空泡变性与血管改变有关。
2 bcl-2/bax基因表达和少突胶质细胞凋亡在海洛因海绵状白质脑病发病机制中的作用研究
2.1研究目的
检测HSLE脑组织细胞凋亡及其调节因子,研究HSLE少突胶质细胞凋亡的发病机制
2.2研究方法
4例HSLE尸检脑组织,5例正常对照脑组织(来源于非脑部病变死亡尸检病例)。检测大脑白质、小脑白质和胼胝体区TUNEl阳性细胞、caspase-3阳性表达及bcl-2/bax基因表达并计数,统计分析采用SPSS 13.0软件包,各阳性细胞数据用均数±标准差(x~-±s)表示,病例组与对照组不同脑区比较采用重复测量数据的方差分析,多重比较采用LSD方法,相同脑区两组间比较采用独立样本t检验,TUNEL细胞凋亡计数与caspase-3表达之间的关系采用-Pearson相关检验。
2.3研究结果
2.3.1 TUNEL细胞凋亡检测
HSLE患者大脑白质有少量凋亡阳性细胞,小脑白质、胼胝体部位大量细胞核呈棕褐色,深染不均,计数明显高于对照组,与对照组相比差异有显著统计学意义(F=327.913,P=0.000);部位间比较差异有统计学意义(F=68.100,P=0.000),部位间多重比较,小脑白质与胼胝体之间差异无统计学意义(P=0.310),大脑白质与小脑、胼胝体比较差异有统计学意义(P=0.018,P=0.017);两组不同脑区之间交互效应具有统计学意义(F=83.715,P=0.000)。
2.3.2 Caspase-3免疫组化阳性表达
caspase-3阳性细胞广泛存在于HSLE大脑、小脑和胼胝体白质,以病灶中心部位为最明显,在边缘区及相对病变程度轻的额叶,也有少量细胞凋亡。两组间比较差异有统计学意义(F=387.955,P=0.000);部位间比较差异有显著统计学意义(F=215.727,P=0.000),部位间多重比较,小脑白质与胼胝体之间差异无统计学意义(P=0.096),大脑白质与小脑白质(P=0.020)、胼胝体(P=0.022)比较差异有显著统计学意义;TUNEL阳性计数与caspase-3阳性计数之间存在正相关(pearson r=0.924,P=0.000)。脑白质主要由少突胶质细胞形成髓鞘,表明HSLE脑白质存在广泛大量少突胶质细胞凋亡。
2.3.3 Bcl-2基因表达
HSLE与对照组间比较差异无统计学意义(F=0.001,P=0.978),大脑白质、小脑白质及胼胝体部位间比较差异无统计学意义(F=0.001,P=0.999)。
2.3.4 Bax基因表达
两组间比较差异有显著统计学意义(F=11.098,P=0.013),HSLE部位间比较差异有显著统计学意义(F=5.632,P=0.016),部位间多重比较,小脑白质与胼胝体之间差异无统计学意义(P=0.952),大脑白质与小脑白质(P=0.032)、胼胝体(P=0.035)比较差异有显著统计学意义。Bcl-2为促生存家族,bax为促凋亡家族,HSLE脑白质bax表达增高与细胞凋亡有关,Bax高表达使线粒体细胞色素C释放,致使Caspases-9和Caspases-3激活而诱导凋亡,Bax与Bcl-2的相对比值在决定细胞生存或死亡中可能起关键作用。
2.4研究结论
(1)少突胶质细胞凋亡是脱髓鞘的原因;
(2)TNF受体1结合后激活caspase-3导致少突胶质细胞凋亡;
(3)Bcl-2/Bax比值减少促使少突胶质细胞凋亡。
3环己铜二腙诱导大鼠脑白质空泡样变性特征及少突胶质细胞凋亡研究
3.1研究目的
通过探索并建立脑白质海绵状空泡变性模型,进一步研究其病因及发病机制,为HSLE病因及发病机制研究提供新的研究内容和思路。
3.2研究方法
SD大鼠72只,动物分组:搀杂物烟熏组(搀熏组)、搀杂物喂食组(搀喂组)、环己铜二腙组(铜腙组)、生理盐水组(对照组),每组分2周、4周、6周3个时间段,每时间段6只SD大鼠。搀杂物按安定:苯巴比妥:异烟肼:水合硅酸镁=2.5∶30∶100∶200的比例研磨成粉备用。搀杂物混合粉剂染毒量每次25mg/100g体重。铜腙组将环己铜二腙配制成1%混悬液,按10mg/100g体重经口灌胃;对照组予等量生理盐水灌胃。均为1次/日。制备SD大鼠脑白质空泡变性膜型,在相应时间点脑组织取材、固定。
HE染色、MBP免疫组化髓鞘染色检测脑白质脱髓鞘及空泡变性特征;透射电镜观察胼胝体压部超微结构;TUNEL技术及caspase-3免疫组化检测细胞凋亡。
统计学方法:应用SPSS 13.0软件包,TUNEL及caspase-3阳性细胞数据用均数±标准差(x~-±s)表示,各组相同脑区及不同时间点之间进行析因方差分析。
3.3研究结果
3.3.1实验鼠大体脑标本
72例SD大鼠脑表面灰白色,无充血、肿胀,各组大体观察无明显差异。
3.3.2 HE染色
搀熏组、搀喂组、铜腙2、4周组及对照组大、小脑结构正常,白质内未见空泡样改变。铜腙6周组可见大鼠脑白质空泡形成,空泡呈长条形或椭圆形,沿纤维走行,以脑干和枕顶叶白质空泡样改变最明显,血管无明显改变,血管周围无明显空泡,无明显炎细胞浸润,未见坏死区,未见明显胶质细胞增生,大脑灰质神经细胞和小脑灰质Purkinje细胞未见缺失和变性。
3.3.3透射电镜
铜腙4周组大鼠,可见髓鞘排列较紊乱,部分结构松解变性,但无典型脱髓鞘改变,铜腙6周组大鼠脑胼胝体可见髓鞘肿胀,髓鞘板层内有空泡形成。
3.3.4 MBP免疫组化染色
搀熏组、搀喂组、铜腙2、4周组及对照组大鼠脑白质、胼胝体染色均匀,未见组织疏松、空泡形成等改变。铜腙6周组可见大鼠脑白质淡染、稀疏,髓鞘中断或被挤向两侧,出现无着色空泡,以脑干和顶枕叶白质最明显。
3.3.5 TUNEL染色
铜腙2周组脑白质偶见散在细胞凋亡,铜腙4周组可见脑白质较多凋亡阳性细胞,深染不均,铜腙6周组可见脑室周白质、小脑白质、胼胝体及脑干部位大量凋亡细胞,分布在空泡区,各组TUNEL计数比较差异具有显著统计学意义(F=309.934,P=0.000),不同时间之间相比差异具有显著统计学意义(F=163.839,P=0.000),各组不同时间交互效应有显著统计学意义(F=155.270,P=0.000),铜腙组与搀熏组、搀喂组及对照组比较差异均有统计学意义(P=0.000),6周组与2周组、4周组比较均有显著统计学意义(P=0.000),4周组与2周组比较具有显著统计学意义(P=0.000),以铜腙6周组最高,其次为铜腙4周组,其余各组及各时间之间差异无统计学意义。提示铜腙4周组脑白质已出现细胞凋亡,随着时间延长,到6周时细胞凋亡加重。
3.3.6 caspase-3免疫组化
铜腙4周组可见脑白质较多凋亡阳性颗粒,深染不均,铜腙6周组可见皮质下白质、小脑白质、胼胝体及脑干部位大量凋亡细胞,分布在脑白质空泡区,各组之间差异具有显著统计学意义(F=84.018,P=0.000),不同时间之间差异具有显著统计学意义(F=23.611,P=0.000),铜腙组与搀熏组、搀喂组及对照组比较差异均有统计学意义(P=0.000),6周组与2周组、4周组比较均有显著统计学意义(P=0.000),4周组与2周组比较有显著统计学意义(P=0.003),以铜腙6周组最高,其次为铜腙4周组,其余各组及各时间之间差异无统计学意义。提示铜腙4周组脑白质出现细胞凋亡信号,随着时间延长,到6周时细胞凋亡加重。与TUNEL凋亡染色相一致,与HE染色及MBP髓鞘染色形态学损害区相一致。说明细胞凋亡与脑白质海绵状空泡变性有关,细胞凋亡可能为脑白质海绵状空泡变性的原因之一。
3.4研究结论
(1)安定、苯巴比妥、异烟肼、水合硅酸镁混合物烟熏或喂食未致鼠脱髓鞘和空泡变;
(2)海洛因搀杂物未引起SD鼠白质细胞凋亡;
(3)鼠经口摄入铜腙主要损害靶点在CNS少突胶质细胞,其凋亡致空泡样变;
(4)铜腙喂食致SD鼠脑白质脱髓鞘及空泡变性,无血管相关性;
(5)出现细胞凋亡(4W)早于出现脑白质脱髓鞘和空泡变性(6W),细胞凋亡是脱髓鞘及空泡变的先导;
(6)细胞凋亡与脱髓鞘、空泡形成有关,呈时间剂量相关性,随铜腙摄入时间延长、总剂量增加,细胞凋亡增多。
4全文总结
(1)HSLE白质病变区影像学髓鞘明显减少;
(2)HSLE形态学脑白质脱髓鞘和空泡变;
(3)HSLE存在血管痉挛及继发性微循环改变;
(4)HSLE白质空泡变性与血管改变有关;
(5)少突胶质细胞凋亡是脱髓鞘的原因;
(6)TNF受体1结合后激活caspase-3导致少突胶质细胞凋亡;
(7)Bcl-2/Bax比值减少促使少突胶质细胞凋亡;
(8)安定、苯巴比妥、异烟肼、水合硅酸镁混合物烟熏或喂食未致鼠脱髓鞘和空泡变;
(9)海洛因搀杂物未引起SD鼠白质细胞凋亡;
(10)鼠经口摄入铜腙主要损害靶点在CNS少突胶质细胞,其凋亡致空泡样变;
(11)铜腙喂食致SD鼠脑白质脱髓鞘及空泡变性,无血管相关性;
(12)出现细胞凋亡(4W)早于出现脑白质脱髓鞘和空泡变性(6W),细胞凋亡是脱髓鞘及空泡变的先导;
(13)细胞凋亡与脱髓鞘、空泡形成有关,呈时间剂量相关性,随铜腙摄入时间延长、总剂量增加,细胞凋亡增多。
Heroin spongiform leukoencephalopathy(HSLE)is a kind of central nervous system organic diseases with white matter spongiform vacuolar degeneration for the pathological features after snorting heroin.Cerebellar ataxia is the main symptoms. This disease was first reported by Wolters E in 1982 after he studied 47 patients in Amsterdam Netherlands.Within the following two decades,many individual cases were reported in some country.In 2000,Dr Lu Bing-Xun firstly diagnosed and reported 28 cases HSLE patients in Guangdong province of China.This was the first outbreak in Chinese mainland and patient had a history of inhaling heroin vapor with noses.Medical image showed that cerebral and cerebellar white matter had extensive and symmetric changes,which was spongiform vacuoles degeneration of white matte, myelin bases protein(MBP)was increase in the cerebrospinal fluid.Its incidence may be related to the central nervous system demyelination.But specific mechanisms unclear.Single intravenous injection of heroin were not found HSLE incidence. Substantial adulterant has been identified from the retail heroin powder.Such as diazepam,isoniazid,talc powder,etc.HSLE are probably caused by heroin adulterant, rather than the heroin itself.Research has shown that secondary oligodendrocyte apoptosis can be caused by radiation,hypoxia-ischemia.Bcl-2/bax change can direct cause neuronal apoptosis pathway.Cuprizone can cause demyelination of white matter,and significantly distribute the splenium of corpus callosum and post-fornix, but the genu of corpus callosum and rostral cerebellar pebuncle is no obvious demyelination.This study adopt imaging and neuropathological to analyze features of cerebral edema and demyelination of white matter in HSLE,used heroin adulterant and cuprizone to prepare rat model of white matter changes,and compared to HSLE, detect oligodendrocytes apoptosis and bcl-2/bax to study the HSLE possible originate from blood vessel and oligodendrocyte apoptosis pathogenesis.
1.Study of characteristics of Neural Pathology in Heroin spongiform leucoencephalopathy imaging and neuropathological
1.1 Purposes
Through the MRI examination,HE staining and MBP immunohistochemical method,to study and explore HSLE features of vacuolar degeneration and the demyelination.
1.2 Methods
Six cases of patients HSLE(Nanfang Hospital,Southern Medical University). The incidences were after withdrawal.MRI was used to Collect T1WI,T2WI, enhancement,FLAIR,DWI and MRA image,comprehensive analysis localizion, qualitation and cerebral vessels change.
4 cases of brain autopsies samples of HSLE and 5 controllers.The frontal lobes, cerebella and corpus callosum were drawn.Demyelination and vacuolation were detected by HE stained and MBP immunohistochemisty under the light microscope. Ultra-structrue were detected by electron microscope method.
1.3 Results
1.3.1 MRI of HSLE patients
Cerebral cortex of HSLE patients become thinning,white matter expansion,no significant edema surrounding lesions,no significant mass effect.Lesions appear long T1WI,long T2WI.There are cerebellar white matter,splenium of corpus callosum, internal capsule posterior branch and occipital white matter lesions in 6 patients. FLAIR showed high signal lesions,like than T2WI slightly reduced,suggest myelination decrease.Enhanced no enhancement,refer to the blood-brain barrier without damage.MRA showed internal carotid artery,anterior cerebral artery,middle cerebral artery,posterior cerebral artery running normal,the arterial trunk slender, some rigid,branch red,and in direct proportion to the severity of clinical symptoms.
1.3.2 Electron microscop
HSLE Oligodendrocytes show multiple vacuolar degenerate.Swollen mitochondria and endoplasmic reticulum can be seen in their reliquus cytoplasm.
1.3.3 HE staining
The encephalon white matter of HSLE presented obvious rarefaction and unequal size vacuole changes.Vacuoles run along the axon.The vacuole changed more from superficial layer to deep layer.The structures were heterocentric.Lower white matter of cerebellar granule show larger vacuoles,to the deeper white matter vacuoles gradually small and intensive.Perivascular Vacuolating were more obvious. There is a small amount of vacuolated changes surrounding the small blood vessels and the capillary.
1.3.4 MBP immunohistochemisty
Compare with the control group,cerebral White matter,cerebellar white matter and corpus callosum showing stained slight and uneven,vacuolated areas no staining, There were growing heavier myelin fiber collapse and fracture from superficial layer of white matter to deep layer.HSLE cerebral cortex can be seen a small amount of filament-like fibers,dyeing uniformity,compared with the control group no significant difference.
1.4 Conclusions
(1)Myelination were obvious decreased in HSLE white matter vacuolar area;
(2)HSLE white matter exist Extensive demyelination and vacuolar degeneration;
(3)Secondary vascular changes increase the vacuolar degeneration.We conjecture cerebral microvascular changes related to vacuole formation.
2 Research of bcl-2/bax gene expression and oligodendrocyte apoptosis in the heroin spongiform leukoencephalopathy pathogenesis
2.1 Purposes
Detect cells apoptosis and its regulatory factors in HSLE brain tissue,research the role of oligodendrocyte apoptosis in HSLE pathogenesis.
2.2 Methods
4 cases of brain autopsies samples of HSLE and 5 controllers.The frontal lobes, cerebella and corpus callosum were drawn.Caspase-3 and bcl-2/bax were stained with immunohistochemical method.TUNEL cells apoptosis were detected under the light microscope.Quantitative analysis was conducted by micro-image analysis instrument.Statistical analysis use SPSS 13.0 software package.Data of the positive cells write as(x~-±s).Comparison of different brain regions in HSLE and control group used repeated measure analysis of variance,multiple comparisons using LSD method,the same brain area comparison between the two groups using independent sample t test,TUNEL apoptotic cell counts and expression of caspase-3 used the Pearson correlation test.
2.3 Results
2.3.1 TUNEL positive counts
There were oligodendrocyte apoptosis in white matter HSLE and apoptotic bodies obviously in vacuolar area,which appeared coloration and partial condense of caryon.Count was significantly higher than control group,compared with the control group the difference was statistical significance(F=327.913,P=0.000);site to compare statistically significant differences(F=68.100,P=0.000),multiple comparisons between sites,cerebellar white matter and corpus callosum was no significant difference(P=0.310),cerebral gray matter and cerebellar white matter,the corpus callosum have statistically significant difference(P=0.018,P=0.017). Different brain regions between two groups there were significant interaction effect (F=83.715,P=0.000).
2.3.2 Caspase-3 immunohistochemisty positive expression
There were leonine Caspase-3 protein positive oligodendrocyes in the focal zone of HSLE,Counts was significantly higher than control group,compared with the control group the difference was statistical significance(F=387.955,P=0.000); Comparison at sites statistically significant differences(F=215.727,P=0.000), multiple comparisons between sites,cerebellar white matter and corpus callosum was no significant difference(P=0.096),cerebral gray matter and cerebellar white matter, the corpus callosum have statistically significant difference(P=0.020,P=0.022). There were direct correlation between TUNEL-positive count and caspase-3 positive counts of HSLE(pearson correlation coefficient r=0.924,P=0.000).Myelins were mainly formed by oligodendrocyte.This indicate HSLE exist extensive white matter oligodendrocyte apoptosis.
2.3.3 Bcl-2 gene expression
The difference was not significant between HSLE and the control group (F=0.001,P=0.978),brain gray matter,white matter and corpus callosum to compare the difference was not significant(F=0.003,P=0.999).
2.3.4 Bax gene expression
The differences of HSLE comparison with the control group were statistical significant(F=11.098,P=0.013).Inter-site difference was significant statistical significance(F=5.632,P=0.016).Multiple sites comparison,cerebellar white matter and corpus callosum was no significant difference(P=0.952),cerebral gray matter and cerebellar white matter(P=0.032)and corpus callosum(P=0.035)difference was significant statistical significance.Bcl-2 is promote the survival family.Bax is the pro-apoptotic family.Bax high expression increase the mitochondrial cytochrome C release,resulting in Caspases-9 and Caspases-3 activation and induced cells apoptosis. Bcl-2 and Bax ratio may play a key role in determining the cell survival or death.
2.4 Conclusions
(1)A large number of apoptotic cells in white matter vacuolation areas,speculated oligodendrocyte apoptosis are probably one of HSLE pathogenesis;
(2)Bcl-2/bax expression change are one of oligodendrocyte apoptosis path;
(3)These were related to Oligodendrocyte poisoning and mitochondrial dysfunction.
3 Characteristics of vacuolar degeneration and oligodendrocyte apoptosis in alba of SD rats with cuprizone induced
3.1 Purposes
Through explore and the establish alba vacuolar degeneration model,further study its etiology and pathogenesis,for etiology and pathogenesis of HSLE study provides new research content and ideas.
3.2 Methods
72 Sprague-Dawley(SD)rats.Rats were randomly divided into four groups: adulterant smoking group(AS),adulterant feeding group(AF),cuprizone feeding group(Cu),and control saline feeding group(con).Each group set 3 time point:2,4 and 6 weeks.We adopt heroin adulterant(INH,diazepam,barbitone and talcum powder)and cuprizone induce SD rats leukoencephalopathy model.Adulterant by ratio:diazepam:isoniazid:barbitone:talcum powder=2.5:30:100:200 mix grinding into powder.Adulterant mixed powder exposed to the volume of each rat by weight 25mg/100g respectively.1%cuprizone were fed rats(10mg/100g weight).While the control group was fed with equal saline.Once a day.
The rats' brains were detected with HE staining and MBP immunohistochemisty staining.Cells apoptosis was counted by TUNEL and caspase-3 immunohistochemisty staining under light microscope,ultra-structure was observed by electron microcope.
Statistical method:Apply SPSS 13.0 software package.The positive counts data of TUNEL and caspase-3 of SD rat model as(x~-±s).Comparision between the same brain regions in each group and of different time points used factorial variance analysis respectively.
3.3 Results
3.3.1 The whole brain samples of SD rats
72 cases of SD rat brain were not seen congestive and swelling.Rats of Cuprizone 6 weeks has mild adhesions between the meninges and the skull.Cerebral, cerebellar gray whiter matter structure clear,no expansion of white matter.
3.3.2 HE staining
Brain gray matter and white matter no vacuolation in AS group,AF groups,Cu 2w,4w group and Con group.Rats of Cu 6w group show the white matter vacuolization,vacuoles were bar shape and running along the fibers.The brain stem and occipital lobe white matter vacuolation were the most obvious,no significant changes in blood vessels,no obvious inflammatory cell infiltration,no necrosis,no obvious glial cell hyperplasia,cerebral gray matter nerve cells were not loss and degeneration.
3.3.3 Electron microscop
Cu 4w group of rats show more disordered arrangement of myelin,but no typical demyelination.Cu 6w group rats corpus callosum can be seen myelin swelling and myelin lamellar Vacuolating.
3.3.4 MBP immunohistochemical
White matter was sparse in Cu 6w group rats.Myelin lamellar split, vacuolization without coloring.The brain stem and top-occipital white matter most obvious.
3.3.5 TUNEL positive cells
More positive apoptotic nuclei were seen in white matter of Cu 4w group,deeply stained unevenly.Large number of apoptotic cells were seen in Cu 6w group,which main located in vacuole area of the ventricle white matter,cerebellar white matter, corpus callosum and brainstem.TUNEL counts in each group the difference was significant statistical significance(F=309.934,P=0.000).The difference between different times was significant statistical significance(F=163.839,P=0.000),Cu 6w group and the 2w,4w group were significantly statistical significance(P=0.000),4w group compared with the 2w group had significant statistical significance(P=0.000), with Cu 6w group were the highest,followed by Cu 4w group.prompted Cu 4w groups have emerged apoptosis in white matter,extended over time,to 6w the apoptosis increase.
3.3.6 Caspase-3 positive expression
This result and TUNEL were accordant.Rats brain gray matter in each group there is a small amount of caspase-3 positive expression,the difference was not significant(F=0.485,P=0.694).Cu 4w group revealed higher apoptosis positive granules.large number of apoptotic cells were count in Cu 6w group.The difference of Caspase-3 expression in each group was significant statistical significance (F=84.018,P=0.000).The differences between different time have significant statistical significance(F=23.611,P=0.000),Cu 6w group and 2w group,4w group were significantly statistical significance(P=0.000),4w group compared with 2w group have significant statistical significance(P=0.003),while Cu 6w group were the highest,followed by Cu 4w group.Prompt Cu 4w group white matter appeared apoptotic signal,apoptosis increaseas by 6w.Cells apoptosis is consistent with myelin lesion morphology of HE staining and MBP staining,and it is earlier than morphology change.Description apoptosis cause white matter vacuolar degeneration. Cell apoptosis may be the originating causes of vacuolar degeneration in white matter.
4 Conclusions
(1)Smoking or feeding rats can not induce SD rats alba white matter changes by diazepam,isoniazid,barbitone and talcum powder adulterant;
(2)Cuprizone oral perfusion may cause SD rats alba vacuolar degeneration;
(3)There were a large number of apoptosis in alba lesions of SD rats by cuprizone induced.Suggesting that apoptosis was related to alba demyelination and vacuolar degeneration;
(4)Damage target of cuprizone were in the central nervous system oligodendrocytes. The oligodendrocyte apoptosis was earlier than alba demyelination at the happened time.Guess apoptosis cause demyelination and vacuolar degeneration in white matter of SD rats by cuprizone induced;
(5)Vacuolation was induced by cuprizone in SD rats.Vacuolating time variability were dose-related,with the time prolonged,the total dose increase,demyelination aggravate;
(6)Cuprizone was used to induced white matter degeneration,no significant brain edema,no obvious vascular relevance;
(7)The cuprizone-induced white matter degeneration were different with HSLE incidence reasons,Neuropathological changes were different,but there are cell apoptosis.
Summary of The full text
1、Myelination were obvious decreased in HSLE white matter vacuolar area;
2、HSLE white matter exist Extensive demyelination and vacuolar degeneration;
3、Secondary vascular changes increase the vacuolar degeneration.We conjecture cerebral microvascular changes related to vacuole formation.
4、A large number of apoptotic cells in white matter vacuolation areas,speculated oligodendrocyte apoptosis are probably one of HSLE pathogenesis;
5、Bcl-2/bax expression change are one of oligodendrocyte apoptosis path;
6、These were related to Oligodendrocyte poisoning and mitochondrial dysfunction;
7、Smoking or feeding rats can not induce SD rats alba white matter changes by diazepam,isoniazid,barbitone and talcum powder adulterant;
8、Cuprizone oral perfusion may cause SD rats alba vacuolar degeneration;
9、There were a large number of apoptosis in alba lesions of SD rats by cuprizone induced.Suggesting that apoptosis was related to alba demyelination and vacuolar degeneration;
10、Damage target of cuprizone were in the central nervous system oligodendrocytes.The oligodendrocyte apoptosis was earlier than alba demyelination at the happened time.Guess apoptosis cause demyelination and vacuolar degeneration in white matter of SD rats with cuprizone induced;
11、Vacuolation were induced by cuprizone in SD rats.Vacuolating time variability were dose-related,with the time prolonged,the total dose increase, demyelination aggravate;
12、Cuprizone were used to induced white matter degeneration,slight cells swelling,no obvious vascular relevance;
13、The cuprizone-induced white matter degeneration were different with HSLE incidence reasons,Neuropathological changes were different,but there are cell apoptosis.
引文
[1]Wolters EC,van Wijngaarden GK,Stam FC,et al.Leucoencephalopathy after inhaling“heroin”pyrolysate[J].Lancet,1982,2(8310):1233-1237.
[2]Nuytten D,Wyffels E,Michiels K,et al.Drug-induced spongiform leucoencephalopathy,a case report with review of the literature[J].Acta Neurol Belg 1998,98(1):32-35.
[3]Weber W,Henkes H,Moller P,et al.Toxis spongiform leucoencephalopathy after inhaling heroin vapour[J].Fur Radiol 1998,8(5):749-755.
[4]Rizzuto N,Morbin M,Ferrari S,et al.Delayed spongiform leukoencephalopathy after heroin abuse[J].Acta Neuropathol(Berl)1997,94(1):87-90.
[5]Chen CY,Less KM,Lee CC,et al.Heroin induced spongiform leukoencephalpathy:value of diffusion MR imaging.[J].comput Assist Tomogr,2000,24:735.
[6]Keogh CF,Andrews GT,Spacey SD,et al.Neuroimaging features of heroin inhalation toxicity:“Chasing the Dragon”[J].AJR Am J Roentgenol.2003Mar,180(3):847-850.
[7]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查[J].中国药物滥用防治杂志.2002,37(2):32-35.
[8]张瑛,苗玲,钱可久,等.海洛因海绵状白质脑病(附2例报道)[J].卒中与神经疾病.2004,11(1):47-49.
[9]冯慧宇,徐评议,张成海,等.洛因海绵状白质脑病的临床特点及治疗分析[J].中华神经科杂志.2006,39(2):130-131.
[10]陆兵勋,周亮,潘速跃等.海洛因海绵状白质脑病的临床和病理特点[J].中华神经科杂志.2001,34(6):347-351.
[11]周亮 陆兵勋 尹恝,等.海洛因海绵状白质脑病的CT和MR表现[J].中华放射学杂志2002,36(1):29-31.
[12]马湘乔,张雪林,张玉忠,等.海洛因脑病的MRI表现[J].中国医学影像技术.2005,21(1):62-64.
[13]董加政,褚晓凡,古坤意,等.烫吸海洛因致白质脑病2例报告及文献复习[J].中风与神经疾病杂志,2002,19(2):108-110.
[14]陆兵勋,王为民,周亮,等.海洛因海绵状白质脑病的影像学特征.临床神经病学杂志[J].2002,15(1):12-14.
[15]王群,刘继元,陆兵勋.海洛因海绵状白质脑病的脑血流动力学变化.中华核医学杂志[J].2002,22(3):156.
[16]黄涛,林丽珍,侯光男,等.海洛因中毒所致海绵状白质脑病患者的神经电生理检测.临床神经电生理学杂志[J].2001,10(2):87-89.
[17]尹恝,陆兵勋,李伟,等.海洛因海绵状白质脑病的脑脊液和血清髓鞘碱性蛋白及其抗体检测及意义中风与神经疾病杂志[J].2003,20(3):222-224.
[18]L Zhou,BX Lu,J Yin,et al.Glucocortioid treatment for heroin-induced spongiform leucoencephalopathy:a clinical controlled study[J].Di Yi Jun Yi Da Xue Xue Bao.2003,23(2):172-174.
[19]刘君,王春雪,牛松涛,等.海洛因海绵状白质脑病的临床及影像学探讨(附1例报告)[J].首都医科大学学报,2005,26(4):502-504.
[20]李才明,张成,刘卫彬.海洛因海绵状白质脑病四例报告[J].中国神经免疫学和神经病学杂志,2005,12(1):56-57.
[21]H Hungerbuhler,W Waespe.Leukoencephalopathy following inhalation of heroin pyrolysate[J].Schweiz Med Wochenschr.1990,120(48):1801-5.
[22]TP Tan,PR Algra,J Valk,and EC Wolters.Toxic leukoencephalopathy after inhalation of poisoned heroin:MR findingsfJ].AJNR Am.J.Neuroradiol.1994,15:175-178.
[23]J Hagel,G Andrews,T Vertinsky,et al.“Chasing the dragon”--imaging of heroin inhalation leukoencephalopathy[J].Can Assoc Radiol.2005,56(4):199-203.
[24]Ciaran F.Keogh,Gordon T.Andrews,Sian D.Spacey,el at.Neuroimaging Features of Heroin Inhalation Toxicity:“Chasing the Dragon”[J].American journal of roentgenology(AJR) 2003,180:847-850.
[25]Kriegstein AR,Shungu DC,Millar WS,et al.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation[J].Neurology,1999,53:1765-1773.
[26]YJ Chang,CH Tsai,CJ Chen.Leukoencephalopathy after inhalation of heroin vapor[J].J Formos Med Assoc.1997,96(9):758-60.
[27]S Vella,R Kreis,KO Lovblad,et al.Acute leukoencephalopathy after inhalation of a single dose of heroin[J].Neuropediatrics,2003,34(2):100-4.
[28]CY Chen,KW Lee,CC Lee,et al.Heroin-induced spongiform leukoencephalopathy:value of diffusion MR imaging[J].J Comput Assist Tomogr,2000,24(5):735-7.
[29]A Ryan,F M Molloy,M A Farrell,et al.Fatal toxic leukoencephalopathy:clinical,radiological,and necropsy findings in two patients[J]Journal of Neurology Neurosurgery and Psychiatry 2005,76:1014-1016.
[30]E Bartlett,D J Mikulis.Chasing “chasing the dragon”with MRI:leukoencephalopathy in drug abuse[J].British Journal of Radiology,2005,78:997-1004.
[31]C Offiah,E Hall.Heroin-induced leukoencephalopathy:characterization using MRI,diffusion-weighted imaging,and MR spectroscopy[J].Clin Radiol,2008,63(2):146-52.
[32]ChangYJ,Tsal CH,Chen CJ.Leukoencephalopathy after inhalation of heroin vapor[J].J FormosMed Assoc,1997,96:758-760.
[33]PJ Egan,FW Becker,F Schumm.Spongiform leucoencephalopathy after inhaling illicit heroin and due to carbon monoxide-intoxication[J].Fortschr Neurol Psychiatr.2004,72(1):26-35.
[34]Buttner A,Mall G,Penning R,et al.The neuropathology of heroin abuse [J].Forensic Sci Int,2000,113:435-442.
[35]A Gacouin,S Lavoue,T Signouret,et al.Reversible spongiform leucoencephalopathy after inhalation of heated heroin[J].Intensive Care Med,2003,29(6):1012-1015.
[36]Gacouin,S Lavoue,T Signouret,et al.Reversible spongiform leucoencephalopathy after inhalation of heated heroin[J].Intensive Care Med.2003,29(6):1012-1015.
[37]Elius EG,Andersson S.Leucoencephalopathy after inhalation of heroin:a case report[J].J Neurol Neurosurg Psychiatry,1996,60:694-695.
[38]Hedley-Whyte ET.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation[J].Neurology,2000,54:2027-2028.
[39]Weber W,Henkes H,Moller P,et al.Toxic spongiform leucoencephalopathy after inhaling heroin vapour[J].Eur Radiol,1998,8:749-755.
[40]Filley CM.Toxic leukoencephalopathy[J].Clin Neuropharmacol.1999,22:249-260.
[41]陆兵勋,马烈,周亮.海洛因海绵状白质脑病流行病学调查[J].中华内科杂志,2001,40:802-805.
[42]陆兵勋,周亮,尹恝,等.海洛因海绵状白质脑病的临床和病理(附一例报告)[J].第一军医大学学报,2000,20:333-335.
[43]陆兵勋,周亮,潘速跃,等.中国海洛因海绵状白质脑病的临床和病理特点[J].中华内科杂志,2001,40:753-756.
[44]戚晓昆.海洛因白质脑病的临床研究进展[J].北京医学,2003,25:275-276.
[45]Romero A A,Gross S R,Cheng K Y,et al.An Age-related Increase in Resistance to DNA Damage-induced Apoptotic Cell Death is Associated with Development of DNA Repair Nbchanisms[J].Neurochem.2003,84(6):1275.
[46]Vartanian T,Li Y,Zhao M,et al.Interferon-gamma-induced oligodendrocyte cell death:implications for the pathogenesis of multiple sclerosis[J].M of Med,1995,1(7):732-743.
[47]Ye P,D'Ercole AJ.Insulin-hlce growth factor Ⅰ protects oligodendrocytes from turn or necrosis factor-alpha-induced injury[J].Endocrinology,1999,140(7):3063-3072.
[48][48]Bonetti B,Raine CS.Multiple sclerosis:oligodendrocytes display cell death-related molecules in situ but do not undergo apoptosis[J].Ann Neurol,1997,42(1):74-84.
[49]Tomimoto H,Ihara M,Wakita H,et al.Chronic cerebral hypoperfusion induces white matter lesions and loss of oligodendroglia with DNA fragmentation in the rat[J].Acta Neuropathol(Berl.2003,106(6):527-534.
[50]Mabuchi T,Kitagawa K,Ohtsaki T,et al.Contribution of microglia/macrophages to expansion of infarction and response of oligodendrocytes after focal cerebral ische mia in rats[J].Stroke.2000,31(7):1735-1743.
[51]Shibata M,Hisahara S,Hara H,et al.Caspases determine the vulnerability of oligodendrocytes in the ischemic brain[J].J Clin Invest.2000,106(5):643-653.
[52]Masumura M,Hata R,Nagai Y,et al.Oligodendroglial cell death with DNA fragmentation in the white matter under chronic cerebral hypoperfusion:comparison between normotensive and spontaneousIy hypertensive rats[J].Neurosci Res.2001,39(4):401-412.
[53]Li GL,Brodin G,Farooque M,et al.Apoptosis and expression of Bcl-2 after compression trauma to rat spinal cord[J].J Neuropathol Exp Neurol.1996,55:280-289.
[54]Tamura M,Nakamura M,Ogawa Y,et al.Targeted expression of anti-apoptotic protein p35 in oligodendrocytes reduces delayed demyelination and functional impairment after spinal cord injury[J].Glia.2005,51(4):312-321.
[55]Azari MF,Profyris C,Karnezis T,et al.Leukemia inhibitory factor arrests oligodendrocyte death and demyelination in spinal cord injury[J].J Neuropathol Exp Neurol.2006,65(9):914-929.
[56]Chiang C S,McBride W H,Wither H R.Radiatiorr induced Astrocytic and microglial Responses in Mouse Brain[J].Radiother Oncol.1993,29:60.
[57]Komoly S.Experimental demyelination caused by primary oligodendrocyte dystrophy.Regional distribution of the lesions in the nervous system of mice [correction][J].Ideggyogy Sz.2005,58(1-2):40-43.
[58]Osterhout DJ,Marin-Husstege M,Abano P,et al.Molecular mechanisms of enhanced susceptibility to apoptosis in differentiating oligodendrocytes[J].J Neurosci Res.2002,69(1):24-29.
[59]Hockenbery D,Oltvai ZN,Yin X,et al.Bcl-2 functions in an an tioxidant pathway to prevent apoptosis[J].Cell,1993,75:241-251.
[60]Sulejczak D,Czarkowska-Bauch J,Macias M,et al.Bcl-2 and Bax proteins are increased in neocortical but not in thalamic apoptosis following devascularizing lesion of the cerebral cortex in the rat:an immunohistochemical study[J].Brain Res.2004,1006(2):133-149.
[61]Hongxin Dong,Alicia Fazzaro,Chuanxi Xiang,et al.Enhanced Oligodendrocyte Survival after Spinal Cord Injury in Bax-Deficient Mice and Mice with Delayed Wallerian Degeneration[J].The Journal of Neuroscience.2003,23(25):8682-8691.
[62]Nesic-Taylor O,Cittelly D,Ye Z.Exogenous Bcl-xL fusion protein spares neurons after spinal cord injury[J].J Neurosci Res.2005,79(5):628-637.
[63]Heather A.Arnett,Ron P.Hellendall,Glenn K.et al.The protective role of nitric oxide in a Neurotoxicant-induced demyelinating Model[J].The Journal of Immunology,2002,168:427-433.
[64]Komoly S,Jeyasingham MD,Pratt OE,et al.Decerase in oligodendrocyte carbonic anhydrase activity preceding myelin degeneration in cuprizone induced demyelination[J].J Neurol Sci.1987,79(1-2):141-8.
[1]Wolters EC,van Wijngaarden GK,Stam FC,et al.Leucoencephalopathy after inhaling“heroin”pyrolysate[J].Lancet,1982,2(8310):1233-1237.
[2]Nuytten D,Wyffels E,Michiels K,et al.Drug-induced spongiform leucoencephalopathy,a case report with review of the literature[J].Acta Neurol Belg 1998,98(1):32-35.
[3]Weber W,Henkes H,Moller P,et al.Toxic spongiform leucoencephalopathy after inhaling heroin vapour[J].Eur Radiol 1998,8(5):749-755.
[4]Rizzuto N,Morbin M,Ferrari S,et al.Delayed spongiform leukoencephalopathy after heroin abuse[J].Acta Neuropathol(Befl)1997,94(1):87-90.
[5]陆兵勋,周亮,潘速跃,等.海洛因海绵状白质脑病的临床和病理特点.[J].中华神经科杂志,2001,34(6):347-350.
[6]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查[J].中国药物滥用防治杂志.2002,37(2):32-35.
[7]张瑛,苗玲,钱可久,等.海洛因海绵状白质脑病(附2例报道)[J].卒中与神经疾病.2004,11(1):47-49.
[8]冯慧宇,徐评议,张成海,等.洛因海绵状白质脑病的临床特点及治疗分析[J].中华神经科杂志.2006,39(2):130-131.
[9]D.H.Chui,E.Dobo,T.Makifuchi.et al.Apoptotic neurons in Alzheimer's disease frequently show intracellular Aβ42 labeling[J].Journal of Alzheimer's Disease,2001,3:231-239.
[10]Takeshi Tabira,De-Hua Chui,Masumi Endoh.The role of MHC class Ⅱ-positive cells in Alzheimer's disease brain[J].Frontiers of Alzheimer Research,Edited by Ishii T,Allsop D,Selkoe DJ,Amsterdam,Elsevier,1991,pp 251-258.
[11]De-hua chui,Takeshi Tabira,Shinzo Izumi,et al.Decreased β-Amylioid and Increased Abnormal Tau Deposition in the Brain of Aged Patients with Leprosy[J].American Journal of Pathology,1994,145(4):771-775.
[12]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查[J].中国药物滥用防治杂志.2002,37(2):32-35.
[13]Strang J,Gurling H.Computerized tomography and neuropsychological assessment in long-term high dose heroin addicts[J].Br J Addict,1989,84:1011-1019.
[14]陆兵勋,马列,周亮,等。海洛因海绵状白质脑病流行病学调查[J].中华内科杂志,2001;40(12):802-805.
[15]Hedley-Whyte ET.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation[J].Neurology.2000,54:2027-2028.
[16]Kriegstein AR,Shungu DC,Millar WS,et al.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation[J].Neurology.1999,53:1765-1773.
[17]N Rizzuto,M Morbin,S Ferrari,et al.Delayed spongiform leukoencephalopathy after heroin abuse[J].Acta Neuropathol.1997,94(1):87-90.
[18]Singh N,Bonham A,Fukui M.Immunosuppressive-associated leukoencephalopathy in organ transplant recipients[J].Transplantation,2000,69:467.
[19]Pirzada NA,Ali Ⅱ,Dafer RM.Fluorouracil-induced neurotoxicity[J].Ann Pharmacother,2000,34:35.
[20]魏万里.无锡地区150例海洛因样品在那个掺杂物检验与讨论[J].中国药物滥用防治杂志.2000,3:26
[21]Buttner A,Mall G,Penning R,et al.The neuropathology of heroin abuse.[J].Forensic Sci Int,2000,113:435-442.
[22]Volkow ND,Valentine A,Kulkarni M,et al.Radiological and neurological changes in the drug abuse patient:a study with MRI[J].J Neuroradiol,1988,15:288-293.
[23]Schoser BG,Groden C.Subacute onset of oculogyric crisis and generalized dystonia following intranasal administration of heroin[J].Addiction,1999,94:431-434.
[24]CY Chen,KW Lee,CC Lee,et al.Heroin-induced spongiform leukoencephalopathy:value of diffusion MR imaging[J].J Comput Assist Tomogr,2000,24(5):735-737.
[25]王光彬,单瑞芹,赵斌,等.脑后部可逆性脑病综合征的CT,MRI诊断.中华放射学杂志,2006,40(9):508-512.
[26]王群,刘继元,陆兵勋.海洛因海绵状白质脑病的脑血流动力学变化.中华核医学杂志[J].2002,22(3):156.
[27]Frei E,Klusman I,Schnell L,et al.Reactions of oligodendrocytes to spinal cord injury:cell survival and myelin repair[J].Exp Neurol,2000,163:373.
[28]Skoff RP,Bessert DA,Barks JD,et al.Hypoxic-ischemic injury results in acute disruption of myelin gene expression and death of oligodendroglial precursors in neonatal mice[J].Int J Devl Neuroscience,2001,19:197.
[29]E Bartlett,D J Mikulis.Chasing“chasing the dragon”with MRI:leukoencephalopathy in drag abuse[J].British Journal of Radiology,2005,78:997-1004.
[30]C Offiah,E Hall.Heroin-induced leukoencephalopathy:characterization using MRI,diffusion-weighted imaging,and MR spectroscopy[J].Clin Radiol,2008,63(2):146-52.
[31]尹恕,陆兵勋,李伟,等.海洛因海绵状白质脑病的脑脊液和血清髓鞘碱性蛋自及其抗体检测及意义.中风与神经疾病杂志.[J].2003,20(3):222-224.
[1]周亮,陆兵勋,尹恝,等.海洛因海绵状白质脑病的CT和MR表现.中华放射学杂志.[J].2002,36(1):29-31.
[2]尹恕,陆兵勋,李伟,等.海洛因海绵状白质脑病的脑脊液和血清髓鞘碱性蛋自及其抗体检测及意义.中风与神经疾病杂志.[J].2003,20(3):222-224.
[3]Romero A A,Gross S R,Cheng K Y,et al.An Age-related Increase in Resistance to DNA Damage-induced Apoptotic Cell Death is Associated with Development of DNA Repair Nbchanisms.[J].Neurochem.2003,84(6):1275.
[4]Chiang C S,McBride W H,Wither H R.Radiatiorr induced Astrocytic and microglial Responses in Mouse Brain.[J].Radiother Oncol.1993,29:60.
[5]Masumura M,Hata R,Nagai Y,et al.Oligodendroglial cell death with DNA fragmentation in the white matter under chronic cerebral hypoperfusion:comparison between normotensive and spontaneously hypertensive rats.[J].Neurosci Res.2001,39(4):401-412.
[6]Osterhout DJ,Marin-Husstege M,Abano P,et al.Molecular mechanisms of enhanced susceptibility to apoptosis in differentiating oligodendrocytes.[J].Neurosci Res.2002,69(1):24-29.
[7]Deluca GC,Nagy Z,Esiri MM,et al.Evidence for a role for apoptosis in central pontine myelinolysis.[J].Acta Neuropatho1(Berl).2002,103(6):590-598.
[8]D.H.Chui,E.Dobo,T.Makifuchi.et al.Apoptotic neurons in Alzheimer's disease frequently show intracellular A|342 labeling.[J].Journal of Alzheimer's Disease,2001,3:231-239.
[9]Takeshi Tabira,De-Hua Chui,Masumi Endoh.The role of MHC class Ⅱ -positive cells in Alzheimer's disease brain.[J].Frontiers of Alzheimer Research,Edited by Ishii T,Allsop D,Selkoe DJ,Amsterdam,Elsevier,1991,pp 251-258.
[10]De-hua chui,Takeshi Tabira,Shinzo Izumi,et al.Decreased P-Amylioid and Increased Abnormal Tau Deposition in the Brain of Aged Patients with Leprosy.[J].American Journal of Pathology,1994,145(4):771-775.
[11]Wolters EC,van Wijngaarden GK,Stam FC,et al.Leucoencephalopathy after inhaling“heroin” pyrolysate.[J].Lancet,1982,2(8310):1233-1237.
[12]Rizzuto N,Morbin M,Ferrari S,et al.Delayed spongiform leukoencephalopathy after heroin abuse.[J].Acta Neuropathol(Befl)1997,94(1):87-90.
[13]Weber W,Henkes H,Moiler P,et al.Toxis spongiform leucoencephalopathy after inhaling heroin vapour.[J].Fur Radiol,1998,8(5):749-755.
[14]AS Robertson,S Jain,RA O'Neil.Spongiform leucoencephalopathy following intravenous heroin abuse:radiological and histopathological findings.[J].Australas Radiol.2001,45(3):390-392.
[15]陆兵勋,周亮,潘速跃,等.海洛因海绵状白质脑病的临床和病理特点.[J].中华神经科杂志,2001,34(6):347-350.
[16]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查.[J].中国药物滥用防治杂志.2002,37(2):32-35.
[17]Elius EG,Andersson S.Leucoencephalopathy after inhalation of heroin:a case report[J].J Neurol Neurosurg Psychiatry.1996,60:694-695.
[18]Buttner A,Mall G,Penning R,et al.The neuropathology of heroin abuse.[J].Forensic Sci Int,2000,113:435-442.
[19]Filley CM.Toxic leukoencephalopathy.[J].Clin Neuropharmacol,1999,22:249-260.
[20]Hacker G.The morphology of apoptosis.[J].Cell Tissue Research,2000,301(1):5-17.
[21]Newrneyer DD,Ferguson-Miller S.Mitochondria:releasing power for life and unleashing the machineries of death.[J].Cell,2003,112(4):481-490.
[22]Thornberry NA,Lazebnik Y.Caspases:enemies within.[J].Science.1998,281:1312-1316.
[23]Breckenridge DG,Xue D.Regulation of mitochondrial membrane permeabilization by BCL-2 family proteins and caspases.[J].Curr Opin Cell Biol 2004,16:647-652.
[24]Wang X,Zhang J,Kim HP,et al.Bcl-XL disrupts death-inducing signal complex formation in plasma membrane induced by hypoxia/reoxygenation.[J].FASEB J 2004,18:1826-1833.
[25]Gabriel B,Sureau F,Casselyn M,et al.Retroactive pathway involving mitochondria in electroloaded cytochrome c-induced apoptosis.Protective properties of Bcl-2 and Bcl-XL.[J].Exp Cell Res 2003,289:195-210.
[26]Pan G,O'Rourke K,Dixit VM.Caspase-9,Bcl-XL,and Apaf-1 form a ternary complex.[J].Biol Chem 1998,273:5841-5845.
[27]Hu Y,Benedict MA,Wu D,et al.Bcl-XL interacts with Apaf-1 and inhibits Apaf-1-dependent caspase-9 activation.[J].Proc Natl Acad Sci USA 1998,95:4386-4391.
[28]Cheng EH,Kirsch DG,Clem RJ,et al.Conversion of Bcl-2 to a Bax-like death effector by caspases.[J].Science.1997,278:1966-1968.
[29]Hongxin Dong,Alicia Fazzaro,Chuanxi Xiang,et al.Enhanced Oligodendrocyte Survival after Spinal Cord Injury in Bax-Deficient Mice and Mice with Delayed Wallerian Degeneration.[J].The Journal of Neuroscience,2003,23(25):8682-8691.
[30]Nesie-Taylor O,Cittelly D,Ye Z.Exogenous Bcl-xL fusion protein spares neurons after spinal cord injury.[J].Neurosci Res,2005,79(5):628-637.
[31]Zhou C,Yamaguchi M,Colohan AR,et al.Role of p53 and apoptosis in cerebral vasospasm after experimental subarachnoid hemorrhage.[J].J Cereb Blood Flow Metab,2005,25(5):572-582.
[32]Adams JM,Cory S.The Bcl-2 protein family:arbiters of cell surviva.[J].Science,1998,281:1322-1326.
[33]Green DR,Kroemer G.The pathophysiology of mitochondrial cell death.[J].Science,2004,305:626-629.
[34]Kriegstein AR,Shungu DC,Millar WS,et al.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation.[J].Neurology.1999,53:1765-1773.
[1]Wolters EC,van Wijngaarden GK,Stam FC,et al.Leucoencephalopathy after inhaling“heroin”pyrolysate[J].Lancet,1982,2:1233-1237.
[2]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查.[J].中国药物滥用防治杂志.2002,37(2):32-35.
[3]Elius EG,Andersson S.Leucoencephalopathy after inhalation of heroin:a case report[J].J Neurol Neurosurg Psychiatry.1996,60:694-695.
[4]尹瑞雪,陆江阳,陆兵勋,等.bcl-2/bax基因表达和少突胶质细胞凋亡在海洛因海绵状白质脑病发病机制中的作用.中华医学杂志.2008,88(11):749-753.
[5]MF Stidworthy,S Genoud,U Surer,et al.Quantifying the early stages of remyelination following cuprizone-induced demyelination.Brain Pathol.2003,13(3):329-339.
[6]S Komoly.Experimental demyelination caused by primary oligodendrocyte dystrophy[J].Regional distribution of the lesions in the nervous system of mice [corrected]Ideggyogy Sz.2005,58(1-2):40-43.
[7]D.H.Chui,E.Dobo,T.Makifuchi,et al.Apoptotic neurons in Alzheimer's disease frequently show intracellular Aβ42 labeling[J].Journal of Alzheimer's Disease,2001,3:231-239.
[8]Takeshi Tabira,De-Hua Chui,Masumi Endoh.The role of MHC class Ⅱ-positive cells in Alzheimer's disease brain[J].Frontiers of Alzheimer Research,Edited by Ishii T,Allsop D,Selkoe D J,Amsterdam,Elsevier,1991,pp 251-258.
[9]De-hua chui,Takeshi Tabira,Shinzo Izumi,et al.Decreased β-Amylioid and Increased Abnormal Tau Deposition in the Brain of Aged Patients with Leprosy.[J].American Journal of Pathology.1994,145(4):771-775.
[10]陆兵勋,周亮,潘速跃等.海洛因海绵状白质脑病的临床和病理特点[J].中华神经科杂志.2001,34(6):347-351.
[11]周亮,陆兵勋,尹恝,等.海洛因海绵状白质脑病的CT和MR表现[J].中华放射学杂志.2002,36(1):29-31.
[12]张瑛,苗玲,钱可久,等.海洛因海绵状白质脑病(附2例报道)[J].卒中与神经疾病.2004,11(1):47-49.
[13]董加政,褚晓凡,古坤意,等.烫吸海洛因致白质脑病2例报告及文献复习[J].中风与神经疾病杂志.2002,19(2):108-110.
[14]H Hungerbuhler,W Waespe.Leukoencephalopathy following inhalation of heroin pyrolysate[J].Schweiz Med Wochenschr.1990,120(48):1801-5.
[15]TP Tan,PR Alga,J Valk,and EC Wolters.Toxic leukoencephalopathy after inhalation of poisoned heroin:MR findings[J].AJNR Am.J.Neuroradiol.Jan 1994,15:175-178.
[16]YJ Chang,CH Tsal,CJ Chen.Leukoencephalopathy after inhalation of heroin vapor[J].J Formos Med Assoc,September 1,1997;96(9):758-60.
[17]Ciaran F.Keogh,Gordon T.Andrews,Sian D.Spacey,el at.Neuroimaging Features of Heroin Inhalation Toxicity:“Chasing the Dragon”[J].American journal of roentgenology(AJR)2003,180:847-850.
[18]Praveen K Gupta,P R Krishnan,P J Sudhakar.Hippocampal involvement due to heroin inhalation-“Chasing the Dragon”[J].Clin Neurol Neurosurg,2008.
[19]Weber W,Henkes H,Moeller P,et al.Toxic spongiform leucoencephalopathy after inhaling heroin vapour[J].Eur Radiol,1998,8(5):749-55.
[20]Filley CM.Toxic leukoencephalopathy[J].Clin Neuropharmacol.1999,22:249-260.
[21]郑志仁,主编.环境病理学.第1版.济南:山东科学技术出版社.1990,56-57
[22]DC Purves,IJ Garrod,AD Dayan,et al.A comparison of spongiosis induced in the brain by hexachlorophene,cuprizone and triethyl tin in the Sprague-Dawley rat[J].Human and Experimental Toxicology.1991,10(6):439-444.
[23]N Franco-Pons,M Torrente,MT Colomina,et al.Behavioral deficits in the cuprizone-induced murine model of demyelination/remyelination[J].Toxicol Lett,2007,169(3):205-13.
[24]SW Sun,HF Liang,K Trinkaus,et al.Noninvasive detection of cuprizone induced axonal damage and demyelination in the mouse corpus callosum[J].Magn Reson Med,2006,55(2):302-8.
[25]QZ Wu,Q Yang,HS Cate,et al.MRI identification of the rostral-caudal pattern of pathology within the corpus callosum in the cuprizone mouse model[J].J Magn Reson Imaging.2008,27(3):446-53.
[26]H Jurevics,C Largent,J Hostettler,et al.Alterations in metabolism and gene expression in brain regions during cuprizone-induced demyelination and remyelination[J].J Neurochem.2002,82(1):126-36.
[27]Strang J,Gurling H.computerized tomography and neuropsychological assessment in long-tern high-dose heroin addicts[J].Br J Addict.1989,84:1011-1019.
[28]Campagnoni AT,Macklin WB.Cellular and molecular aspects of myelin protein gene expression[J].Mol Neurobiol.1988,2:41-89.
[29]Roach A,Takahashi N,Pravtcheva D,el at.Chromosomal mapping of mouse myelin basic protein gene and structure and transcription of the partially deleted gene in shiverer mutant mouse[J].Cell.1985,42:149-155.
[30]Newmeyer DD,Ferguson- Miller S.Mitochondria:releasing power for life and unleashing the machineries of death[J].Cell.2003,112(4):481-490.
[31]Thornberry NA,Lazebnik Y.Caspases:enemies within[XJ.Science.1998,281:1312-1316.
[32]Kuida K,Zheng TS,Na S,et al.Decreased apoptosis in the brain and premature lethalitu in CPP32-deficient mice[J].Nature,1996,384(6607):368-372.
[33]Adam JM,Corys.Apoptosomes:engines for caspase activation[J].Curropin Cell Biol.2002,14(6):715.
[34]Cheng EH,Kirsch DQClem PJ,Ravi R,Kastan MB,Bedi A,Ueno K,Hardwick JM.Conversion of Bcl-2 to a Bax-like death effector by caspases[J].Science.1997,278:1966-1968.
[35]Shi Y.Mechanisms of caspase activation and inhibition during apoptosis[J].MolCel.l 2002,9(3):459.
[36]Romero A A,Gross S R,Cheng K Y,et al.An Age-related Increase in Resistance to DNA Damage-induced Apoptotic Cell Death is Associated with Development of DNA Repair Nbchanisms[J].Neurochem,2003,84(6):1275.
[37]Chiang C S,McBride W H,Wither H R.Radiatiorr induced Astrocytic and microglial Responses in Mouse Brain[J].Radiother Oncol,1993,29:60.
[38]W Cammer.The neurotoxicant,cuprizone,retards the differentiation of oligodendrocytes in vitro[J].J Neurol Sci,October 15,1999,168(2):116-120.
[39]Lipton P.Ischemic cell death in brain neurons[J].Physiol Rev,1999,79(4): 1431.
[40]Kurumatani T,Kudo T,Ikura Y,et al.White matter changes in the Gerbil brain under chronic cerebral hypoperfusion[J].Stroke,1998,29:1058-1062.
[41]Jeffrey L.Mason,Arrel Toews,Janell D.Hostettler,et al.Oligodendrocytes and Progenitors Become Progressively Depleted within Chronically Demyelinated Lesions[J].American Journal of Pathology.2004,164:1673-1682.
[42]Herts SW.To Die orNot to Die:an overview of apoptosis and its role in disease[J].JAMA,1998,279:300-307.
[43]Lavrik IN,GolksA,KrammerPH.Caspases:pharmacological manipulation of cell death[J].J Clin Invest,2005,115:2665-2672.
[2]Nuytten D,Wyffels E,Michiels K,et al.Drug-induced spongiform leucoencephalopathy,a case report with review of the literature[J].Acta Neurol Belg 1998,98(1):32-35.
[3]Weber W,Henkes H,Moller P,et al.Toxis spongiform leucoencephalopathy after inhaling heroin vapour[J].Fur Radiol 1998,8(5):749-755.
[4]Rizzuto N,Morbin M,Ferrari S,et al.Delayed spongiform leukoencephalopathy after heroin abuse[J].Acta Neuropathol(Berl)1997,94(1):87-90.
[5]Chen CY,Less KM,Lee CC,et al.Heroin induced spongiform leukoencephalpathy:value of diffusion MR imaging.[J].comput Assist Tomogr,2000,24:735.
[6]Keogh CF,Andrews GT,Spacey SD,et al.Neuroimaging features of heroin inhalation toxicity:“Chasing the Dragon”[J].AJR Am J Roentgenol.2003Mar,180(3):847-850.
[7]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查[J].中国药物滥用防治杂志.2002,37(2):32-35.
[8]张瑛,苗玲,钱可久,等.海洛因海绵状白质脑病(附2例报道)[J].卒中与神经疾病.2004,11(1):47-49.
[9]冯慧宇,徐评议,张成海,等.洛因海绵状白质脑病的临床特点及治疗分析[J].中华神经科杂志.2006,39(2):130-131.
[10]陆兵勋,周亮,潘速跃等.海洛因海绵状白质脑病的临床和病理特点[J].中华神经科杂志.2001,34(6):347-351.
[11]周亮 陆兵勋 尹恝,等.海洛因海绵状白质脑病的CT和MR表现[J].中华放射学杂志2002,36(1):29-31.
[12]马湘乔,张雪林,张玉忠,等.海洛因脑病的MRI表现[J].中国医学影像技术.2005,21(1):62-64.
[13]董加政,褚晓凡,古坤意,等.烫吸海洛因致白质脑病2例报告及文献复习[J].中风与神经疾病杂志,2002,19(2):108-110.
[14]陆兵勋,王为民,周亮,等.海洛因海绵状白质脑病的影像学特征.临床神经病学杂志[J].2002,15(1):12-14.
[15]王群,刘继元,陆兵勋.海洛因海绵状白质脑病的脑血流动力学变化.中华核医学杂志[J].2002,22(3):156.
[16]黄涛,林丽珍,侯光男,等.海洛因中毒所致海绵状白质脑病患者的神经电生理检测.临床神经电生理学杂志[J].2001,10(2):87-89.
[17]尹恝,陆兵勋,李伟,等.海洛因海绵状白质脑病的脑脊液和血清髓鞘碱性蛋白及其抗体检测及意义中风与神经疾病杂志[J].2003,20(3):222-224.
[18]L Zhou,BX Lu,J Yin,et al.Glucocortioid treatment for heroin-induced spongiform leucoencephalopathy:a clinical controlled study[J].Di Yi Jun Yi Da Xue Xue Bao.2003,23(2):172-174.
[19]刘君,王春雪,牛松涛,等.海洛因海绵状白质脑病的临床及影像学探讨(附1例报告)[J].首都医科大学学报,2005,26(4):502-504.
[20]李才明,张成,刘卫彬.海洛因海绵状白质脑病四例报告[J].中国神经免疫学和神经病学杂志,2005,12(1):56-57.
[21]H Hungerbuhler,W Waespe.Leukoencephalopathy following inhalation of heroin pyrolysate[J].Schweiz Med Wochenschr.1990,120(48):1801-5.
[22]TP Tan,PR Algra,J Valk,and EC Wolters.Toxic leukoencephalopathy after inhalation of poisoned heroin:MR findingsfJ].AJNR Am.J.Neuroradiol.1994,15:175-178.
[23]J Hagel,G Andrews,T Vertinsky,et al.“Chasing the dragon”--imaging of heroin inhalation leukoencephalopathy[J].Can Assoc Radiol.2005,56(4):199-203.
[24]Ciaran F.Keogh,Gordon T.Andrews,Sian D.Spacey,el at.Neuroimaging Features of Heroin Inhalation Toxicity:“Chasing the Dragon”[J].American journal of roentgenology(AJR) 2003,180:847-850.
[25]Kriegstein AR,Shungu DC,Millar WS,et al.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation[J].Neurology,1999,53:1765-1773.
[26]YJ Chang,CH Tsai,CJ Chen.Leukoencephalopathy after inhalation of heroin vapor[J].J Formos Med Assoc.1997,96(9):758-60.
[27]S Vella,R Kreis,KO Lovblad,et al.Acute leukoencephalopathy after inhalation of a single dose of heroin[J].Neuropediatrics,2003,34(2):100-4.
[28]CY Chen,KW Lee,CC Lee,et al.Heroin-induced spongiform leukoencephalopathy:value of diffusion MR imaging[J].J Comput Assist Tomogr,2000,24(5):735-7.
[29]A Ryan,F M Molloy,M A Farrell,et al.Fatal toxic leukoencephalopathy:clinical,radiological,and necropsy findings in two patients[J]Journal of Neurology Neurosurgery and Psychiatry 2005,76:1014-1016.
[30]E Bartlett,D J Mikulis.Chasing “chasing the dragon”with MRI:leukoencephalopathy in drug abuse[J].British Journal of Radiology,2005,78:997-1004.
[31]C Offiah,E Hall.Heroin-induced leukoencephalopathy:characterization using MRI,diffusion-weighted imaging,and MR spectroscopy[J].Clin Radiol,2008,63(2):146-52.
[32]ChangYJ,Tsal CH,Chen CJ.Leukoencephalopathy after inhalation of heroin vapor[J].J FormosMed Assoc,1997,96:758-760.
[33]PJ Egan,FW Becker,F Schumm.Spongiform leucoencephalopathy after inhaling illicit heroin and due to carbon monoxide-intoxication[J].Fortschr Neurol Psychiatr.2004,72(1):26-35.
[34]Buttner A,Mall G,Penning R,et al.The neuropathology of heroin abuse [J].Forensic Sci Int,2000,113:435-442.
[35]A Gacouin,S Lavoue,T Signouret,et al.Reversible spongiform leucoencephalopathy after inhalation of heated heroin[J].Intensive Care Med,2003,29(6):1012-1015.
[36]Gacouin,S Lavoue,T Signouret,et al.Reversible spongiform leucoencephalopathy after inhalation of heated heroin[J].Intensive Care Med.2003,29(6):1012-1015.
[37]Elius EG,Andersson S.Leucoencephalopathy after inhalation of heroin:a case report[J].J Neurol Neurosurg Psychiatry,1996,60:694-695.
[38]Hedley-Whyte ET.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation[J].Neurology,2000,54:2027-2028.
[39]Weber W,Henkes H,Moller P,et al.Toxic spongiform leucoencephalopathy after inhaling heroin vapour[J].Eur Radiol,1998,8:749-755.
[40]Filley CM.Toxic leukoencephalopathy[J].Clin Neuropharmacol.1999,22:249-260.
[41]陆兵勋,马烈,周亮.海洛因海绵状白质脑病流行病学调查[J].中华内科杂志,2001,40:802-805.
[42]陆兵勋,周亮,尹恝,等.海洛因海绵状白质脑病的临床和病理(附一例报告)[J].第一军医大学学报,2000,20:333-335.
[43]陆兵勋,周亮,潘速跃,等.中国海洛因海绵状白质脑病的临床和病理特点[J].中华内科杂志,2001,40:753-756.
[44]戚晓昆.海洛因白质脑病的临床研究进展[J].北京医学,2003,25:275-276.
[45]Romero A A,Gross S R,Cheng K Y,et al.An Age-related Increase in Resistance to DNA Damage-induced Apoptotic Cell Death is Associated with Development of DNA Repair Nbchanisms[J].Neurochem.2003,84(6):1275.
[46]Vartanian T,Li Y,Zhao M,et al.Interferon-gamma-induced oligodendrocyte cell death:implications for the pathogenesis of multiple sclerosis[J].M of Med,1995,1(7):732-743.
[47]Ye P,D'Ercole AJ.Insulin-hlce growth factor Ⅰ protects oligodendrocytes from turn or necrosis factor-alpha-induced injury[J].Endocrinology,1999,140(7):3063-3072.
[48][48]Bonetti B,Raine CS.Multiple sclerosis:oligodendrocytes display cell death-related molecules in situ but do not undergo apoptosis[J].Ann Neurol,1997,42(1):74-84.
[49]Tomimoto H,Ihara M,Wakita H,et al.Chronic cerebral hypoperfusion induces white matter lesions and loss of oligodendroglia with DNA fragmentation in the rat[J].Acta Neuropathol(Berl.2003,106(6):527-534.
[50]Mabuchi T,Kitagawa K,Ohtsaki T,et al.Contribution of microglia/macrophages to expansion of infarction and response of oligodendrocytes after focal cerebral ische mia in rats[J].Stroke.2000,31(7):1735-1743.
[51]Shibata M,Hisahara S,Hara H,et al.Caspases determine the vulnerability of oligodendrocytes in the ischemic brain[J].J Clin Invest.2000,106(5):643-653.
[52]Masumura M,Hata R,Nagai Y,et al.Oligodendroglial cell death with DNA fragmentation in the white matter under chronic cerebral hypoperfusion:comparison between normotensive and spontaneousIy hypertensive rats[J].Neurosci Res.2001,39(4):401-412.
[53]Li GL,Brodin G,Farooque M,et al.Apoptosis and expression of Bcl-2 after compression trauma to rat spinal cord[J].J Neuropathol Exp Neurol.1996,55:280-289.
[54]Tamura M,Nakamura M,Ogawa Y,et al.Targeted expression of anti-apoptotic protein p35 in oligodendrocytes reduces delayed demyelination and functional impairment after spinal cord injury[J].Glia.2005,51(4):312-321.
[55]Azari MF,Profyris C,Karnezis T,et al.Leukemia inhibitory factor arrests oligodendrocyte death and demyelination in spinal cord injury[J].J Neuropathol Exp Neurol.2006,65(9):914-929.
[56]Chiang C S,McBride W H,Wither H R.Radiatiorr induced Astrocytic and microglial Responses in Mouse Brain[J].Radiother Oncol.1993,29:60.
[57]Komoly S.Experimental demyelination caused by primary oligodendrocyte dystrophy.Regional distribution of the lesions in the nervous system of mice [correction][J].Ideggyogy Sz.2005,58(1-2):40-43.
[58]Osterhout DJ,Marin-Husstege M,Abano P,et al.Molecular mechanisms of enhanced susceptibility to apoptosis in differentiating oligodendrocytes[J].J Neurosci Res.2002,69(1):24-29.
[59]Hockenbery D,Oltvai ZN,Yin X,et al.Bcl-2 functions in an an tioxidant pathway to prevent apoptosis[J].Cell,1993,75:241-251.
[60]Sulejczak D,Czarkowska-Bauch J,Macias M,et al.Bcl-2 and Bax proteins are increased in neocortical but not in thalamic apoptosis following devascularizing lesion of the cerebral cortex in the rat:an immunohistochemical study[J].Brain Res.2004,1006(2):133-149.
[61]Hongxin Dong,Alicia Fazzaro,Chuanxi Xiang,et al.Enhanced Oligodendrocyte Survival after Spinal Cord Injury in Bax-Deficient Mice and Mice with Delayed Wallerian Degeneration[J].The Journal of Neuroscience.2003,23(25):8682-8691.
[62]Nesic-Taylor O,Cittelly D,Ye Z.Exogenous Bcl-xL fusion protein spares neurons after spinal cord injury[J].J Neurosci Res.2005,79(5):628-637.
[63]Heather A.Arnett,Ron P.Hellendall,Glenn K.et al.The protective role of nitric oxide in a Neurotoxicant-induced demyelinating Model[J].The Journal of Immunology,2002,168:427-433.
[64]Komoly S,Jeyasingham MD,Pratt OE,et al.Decerase in oligodendrocyte carbonic anhydrase activity preceding myelin degeneration in cuprizone induced demyelination[J].J Neurol Sci.1987,79(1-2):141-8.
[1]Wolters EC,van Wijngaarden GK,Stam FC,et al.Leucoencephalopathy after inhaling“heroin”pyrolysate[J].Lancet,1982,2(8310):1233-1237.
[2]Nuytten D,Wyffels E,Michiels K,et al.Drug-induced spongiform leucoencephalopathy,a case report with review of the literature[J].Acta Neurol Belg 1998,98(1):32-35.
[3]Weber W,Henkes H,Moller P,et al.Toxic spongiform leucoencephalopathy after inhaling heroin vapour[J].Eur Radiol 1998,8(5):749-755.
[4]Rizzuto N,Morbin M,Ferrari S,et al.Delayed spongiform leukoencephalopathy after heroin abuse[J].Acta Neuropathol(Befl)1997,94(1):87-90.
[5]陆兵勋,周亮,潘速跃,等.海洛因海绵状白质脑病的临床和病理特点.[J].中华神经科杂志,2001,34(6):347-350.
[6]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查[J].中国药物滥用防治杂志.2002,37(2):32-35.
[7]张瑛,苗玲,钱可久,等.海洛因海绵状白质脑病(附2例报道)[J].卒中与神经疾病.2004,11(1):47-49.
[8]冯慧宇,徐评议,张成海,等.洛因海绵状白质脑病的临床特点及治疗分析[J].中华神经科杂志.2006,39(2):130-131.
[9]D.H.Chui,E.Dobo,T.Makifuchi.et al.Apoptotic neurons in Alzheimer's disease frequently show intracellular Aβ42 labeling[J].Journal of Alzheimer's Disease,2001,3:231-239.
[10]Takeshi Tabira,De-Hua Chui,Masumi Endoh.The role of MHC class Ⅱ-positive cells in Alzheimer's disease brain[J].Frontiers of Alzheimer Research,Edited by Ishii T,Allsop D,Selkoe DJ,Amsterdam,Elsevier,1991,pp 251-258.
[11]De-hua chui,Takeshi Tabira,Shinzo Izumi,et al.Decreased β-Amylioid and Increased Abnormal Tau Deposition in the Brain of Aged Patients with Leprosy[J].American Journal of Pathology,1994,145(4):771-775.
[12]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查[J].中国药物滥用防治杂志.2002,37(2):32-35.
[13]Strang J,Gurling H.Computerized tomography and neuropsychological assessment in long-term high dose heroin addicts[J].Br J Addict,1989,84:1011-1019.
[14]陆兵勋,马列,周亮,等。海洛因海绵状白质脑病流行病学调查[J].中华内科杂志,2001;40(12):802-805.
[15]Hedley-Whyte ET.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation[J].Neurology.2000,54:2027-2028.
[16]Kriegstein AR,Shungu DC,Millar WS,et al.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation[J].Neurology.1999,53:1765-1773.
[17]N Rizzuto,M Morbin,S Ferrari,et al.Delayed spongiform leukoencephalopathy after heroin abuse[J].Acta Neuropathol.1997,94(1):87-90.
[18]Singh N,Bonham A,Fukui M.Immunosuppressive-associated leukoencephalopathy in organ transplant recipients[J].Transplantation,2000,69:467.
[19]Pirzada NA,Ali Ⅱ,Dafer RM.Fluorouracil-induced neurotoxicity[J].Ann Pharmacother,2000,34:35.
[20]魏万里.无锡地区150例海洛因样品在那个掺杂物检验与讨论[J].中国药物滥用防治杂志.2000,3:26
[21]Buttner A,Mall G,Penning R,et al.The neuropathology of heroin abuse.[J].Forensic Sci Int,2000,113:435-442.
[22]Volkow ND,Valentine A,Kulkarni M,et al.Radiological and neurological changes in the drug abuse patient:a study with MRI[J].J Neuroradiol,1988,15:288-293.
[23]Schoser BG,Groden C.Subacute onset of oculogyric crisis and generalized dystonia following intranasal administration of heroin[J].Addiction,1999,94:431-434.
[24]CY Chen,KW Lee,CC Lee,et al.Heroin-induced spongiform leukoencephalopathy:value of diffusion MR imaging[J].J Comput Assist Tomogr,2000,24(5):735-737.
[25]王光彬,单瑞芹,赵斌,等.脑后部可逆性脑病综合征的CT,MRI诊断.中华放射学杂志,2006,40(9):508-512.
[26]王群,刘继元,陆兵勋.海洛因海绵状白质脑病的脑血流动力学变化.中华核医学杂志[J].2002,22(3):156.
[27]Frei E,Klusman I,Schnell L,et al.Reactions of oligodendrocytes to spinal cord injury:cell survival and myelin repair[J].Exp Neurol,2000,163:373.
[28]Skoff RP,Bessert DA,Barks JD,et al.Hypoxic-ischemic injury results in acute disruption of myelin gene expression and death of oligodendroglial precursors in neonatal mice[J].Int J Devl Neuroscience,2001,19:197.
[29]E Bartlett,D J Mikulis.Chasing“chasing the dragon”with MRI:leukoencephalopathy in drag abuse[J].British Journal of Radiology,2005,78:997-1004.
[30]C Offiah,E Hall.Heroin-induced leukoencephalopathy:characterization using MRI,diffusion-weighted imaging,and MR spectroscopy[J].Clin Radiol,2008,63(2):146-52.
[31]尹恕,陆兵勋,李伟,等.海洛因海绵状白质脑病的脑脊液和血清髓鞘碱性蛋自及其抗体检测及意义.中风与神经疾病杂志.[J].2003,20(3):222-224.
[1]周亮,陆兵勋,尹恝,等.海洛因海绵状白质脑病的CT和MR表现.中华放射学杂志.[J].2002,36(1):29-31.
[2]尹恕,陆兵勋,李伟,等.海洛因海绵状白质脑病的脑脊液和血清髓鞘碱性蛋自及其抗体检测及意义.中风与神经疾病杂志.[J].2003,20(3):222-224.
[3]Romero A A,Gross S R,Cheng K Y,et al.An Age-related Increase in Resistance to DNA Damage-induced Apoptotic Cell Death is Associated with Development of DNA Repair Nbchanisms.[J].Neurochem.2003,84(6):1275.
[4]Chiang C S,McBride W H,Wither H R.Radiatiorr induced Astrocytic and microglial Responses in Mouse Brain.[J].Radiother Oncol.1993,29:60.
[5]Masumura M,Hata R,Nagai Y,et al.Oligodendroglial cell death with DNA fragmentation in the white matter under chronic cerebral hypoperfusion:comparison between normotensive and spontaneously hypertensive rats.[J].Neurosci Res.2001,39(4):401-412.
[6]Osterhout DJ,Marin-Husstege M,Abano P,et al.Molecular mechanisms of enhanced susceptibility to apoptosis in differentiating oligodendrocytes.[J].Neurosci Res.2002,69(1):24-29.
[7]Deluca GC,Nagy Z,Esiri MM,et al.Evidence for a role for apoptosis in central pontine myelinolysis.[J].Acta Neuropatho1(Berl).2002,103(6):590-598.
[8]D.H.Chui,E.Dobo,T.Makifuchi.et al.Apoptotic neurons in Alzheimer's disease frequently show intracellular A|342 labeling.[J].Journal of Alzheimer's Disease,2001,3:231-239.
[9]Takeshi Tabira,De-Hua Chui,Masumi Endoh.The role of MHC class Ⅱ -positive cells in Alzheimer's disease brain.[J].Frontiers of Alzheimer Research,Edited by Ishii T,Allsop D,Selkoe DJ,Amsterdam,Elsevier,1991,pp 251-258.
[10]De-hua chui,Takeshi Tabira,Shinzo Izumi,et al.Decreased P-Amylioid and Increased Abnormal Tau Deposition in the Brain of Aged Patients with Leprosy.[J].American Journal of Pathology,1994,145(4):771-775.
[11]Wolters EC,van Wijngaarden GK,Stam FC,et al.Leucoencephalopathy after inhaling“heroin” pyrolysate.[J].Lancet,1982,2(8310):1233-1237.
[12]Rizzuto N,Morbin M,Ferrari S,et al.Delayed spongiform leukoencephalopathy after heroin abuse.[J].Acta Neuropathol(Befl)1997,94(1):87-90.
[13]Weber W,Henkes H,Moiler P,et al.Toxis spongiform leucoencephalopathy after inhaling heroin vapour.[J].Fur Radiol,1998,8(5):749-755.
[14]AS Robertson,S Jain,RA O'Neil.Spongiform leucoencephalopathy following intravenous heroin abuse:radiological and histopathological findings.[J].Australas Radiol.2001,45(3):390-392.
[15]陆兵勋,周亮,潘速跃,等.海洛因海绵状白质脑病的临床和病理特点.[J].中华神经科杂志,2001,34(6):347-350.
[16]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查.[J].中国药物滥用防治杂志.2002,37(2):32-35.
[17]Elius EG,Andersson S.Leucoencephalopathy after inhalation of heroin:a case report[J].J Neurol Neurosurg Psychiatry.1996,60:694-695.
[18]Buttner A,Mall G,Penning R,et al.The neuropathology of heroin abuse.[J].Forensic Sci Int,2000,113:435-442.
[19]Filley CM.Toxic leukoencephalopathy.[J].Clin Neuropharmacol,1999,22:249-260.
[20]Hacker G.The morphology of apoptosis.[J].Cell Tissue Research,2000,301(1):5-17.
[21]Newrneyer DD,Ferguson-Miller S.Mitochondria:releasing power for life and unleashing the machineries of death.[J].Cell,2003,112(4):481-490.
[22]Thornberry NA,Lazebnik Y.Caspases:enemies within.[J].Science.1998,281:1312-1316.
[23]Breckenridge DG,Xue D.Regulation of mitochondrial membrane permeabilization by BCL-2 family proteins and caspases.[J].Curr Opin Cell Biol 2004,16:647-652.
[24]Wang X,Zhang J,Kim HP,et al.Bcl-XL disrupts death-inducing signal complex formation in plasma membrane induced by hypoxia/reoxygenation.[J].FASEB J 2004,18:1826-1833.
[25]Gabriel B,Sureau F,Casselyn M,et al.Retroactive pathway involving mitochondria in electroloaded cytochrome c-induced apoptosis.Protective properties of Bcl-2 and Bcl-XL.[J].Exp Cell Res 2003,289:195-210.
[26]Pan G,O'Rourke K,Dixit VM.Caspase-9,Bcl-XL,and Apaf-1 form a ternary complex.[J].Biol Chem 1998,273:5841-5845.
[27]Hu Y,Benedict MA,Wu D,et al.Bcl-XL interacts with Apaf-1 and inhibits Apaf-1-dependent caspase-9 activation.[J].Proc Natl Acad Sci USA 1998,95:4386-4391.
[28]Cheng EH,Kirsch DG,Clem RJ,et al.Conversion of Bcl-2 to a Bax-like death effector by caspases.[J].Science.1997,278:1966-1968.
[29]Hongxin Dong,Alicia Fazzaro,Chuanxi Xiang,et al.Enhanced Oligodendrocyte Survival after Spinal Cord Injury in Bax-Deficient Mice and Mice with Delayed Wallerian Degeneration.[J].The Journal of Neuroscience,2003,23(25):8682-8691.
[30]Nesie-Taylor O,Cittelly D,Ye Z.Exogenous Bcl-xL fusion protein spares neurons after spinal cord injury.[J].Neurosci Res,2005,79(5):628-637.
[31]Zhou C,Yamaguchi M,Colohan AR,et al.Role of p53 and apoptosis in cerebral vasospasm after experimental subarachnoid hemorrhage.[J].J Cereb Blood Flow Metab,2005,25(5):572-582.
[32]Adams JM,Cory S.The Bcl-2 protein family:arbiters of cell surviva.[J].Science,1998,281:1322-1326.
[33]Green DR,Kroemer G.The pathophysiology of mitochondrial cell death.[J].Science,2004,305:626-629.
[34]Kriegstein AR,Shungu DC,Millar WS,et al.Leukoencephalopathy and raised brain lactate from heroin vapor inhalation.[J].Neurology.1999,53:1765-1773.
[1]Wolters EC,van Wijngaarden GK,Stam FC,et al.Leucoencephalopathy after inhaling“heroin”pyrolysate[J].Lancet,1982,2:1233-1237.
[2]陆兵勋,马烈,周亮,等.广东沿海地区吸毒人群海洛因海绵状白质脑病流行病学调查.[J].中国药物滥用防治杂志.2002,37(2):32-35.
[3]Elius EG,Andersson S.Leucoencephalopathy after inhalation of heroin:a case report[J].J Neurol Neurosurg Psychiatry.1996,60:694-695.
[4]尹瑞雪,陆江阳,陆兵勋,等.bcl-2/bax基因表达和少突胶质细胞凋亡在海洛因海绵状白质脑病发病机制中的作用.中华医学杂志.2008,88(11):749-753.
[5]MF Stidworthy,S Genoud,U Surer,et al.Quantifying the early stages of remyelination following cuprizone-induced demyelination.Brain Pathol.2003,13(3):329-339.
[6]S Komoly.Experimental demyelination caused by primary oligodendrocyte dystrophy[J].Regional distribution of the lesions in the nervous system of mice [corrected]Ideggyogy Sz.2005,58(1-2):40-43.
[7]D.H.Chui,E.Dobo,T.Makifuchi,et al.Apoptotic neurons in Alzheimer's disease frequently show intracellular Aβ42 labeling[J].Journal of Alzheimer's Disease,2001,3:231-239.
[8]Takeshi Tabira,De-Hua Chui,Masumi Endoh.The role of MHC class Ⅱ-positive cells in Alzheimer's disease brain[J].Frontiers of Alzheimer Research,Edited by Ishii T,Allsop D,Selkoe D J,Amsterdam,Elsevier,1991,pp 251-258.
[9]De-hua chui,Takeshi Tabira,Shinzo Izumi,et al.Decreased β-Amylioid and Increased Abnormal Tau Deposition in the Brain of Aged Patients with Leprosy.[J].American Journal of Pathology.1994,145(4):771-775.
[10]陆兵勋,周亮,潘速跃等.海洛因海绵状白质脑病的临床和病理特点[J].中华神经科杂志.2001,34(6):347-351.
[11]周亮,陆兵勋,尹恝,等.海洛因海绵状白质脑病的CT和MR表现[J].中华放射学杂志.2002,36(1):29-31.
[12]张瑛,苗玲,钱可久,等.海洛因海绵状白质脑病(附2例报道)[J].卒中与神经疾病.2004,11(1):47-49.
[13]董加政,褚晓凡,古坤意,等.烫吸海洛因致白质脑病2例报告及文献复习[J].中风与神经疾病杂志.2002,19(2):108-110.
[14]H Hungerbuhler,W Waespe.Leukoencephalopathy following inhalation of heroin pyrolysate[J].Schweiz Med Wochenschr.1990,120(48):1801-5.
[15]TP Tan,PR Alga,J Valk,and EC Wolters.Toxic leukoencephalopathy after inhalation of poisoned heroin:MR findings[J].AJNR Am.J.Neuroradiol.Jan 1994,15:175-178.
[16]YJ Chang,CH Tsal,CJ Chen.Leukoencephalopathy after inhalation of heroin vapor[J].J Formos Med Assoc,September 1,1997;96(9):758-60.
[17]Ciaran F.Keogh,Gordon T.Andrews,Sian D.Spacey,el at.Neuroimaging Features of Heroin Inhalation Toxicity:“Chasing the Dragon”[J].American journal of roentgenology(AJR)2003,180:847-850.
[18]Praveen K Gupta,P R Krishnan,P J Sudhakar.Hippocampal involvement due to heroin inhalation-“Chasing the Dragon”[J].Clin Neurol Neurosurg,2008.
[19]Weber W,Henkes H,Moeller P,et al.Toxic spongiform leucoencephalopathy after inhaling heroin vapour[J].Eur Radiol,1998,8(5):749-55.
[20]Filley CM.Toxic leukoencephalopathy[J].Clin Neuropharmacol.1999,22:249-260.
[21]郑志仁,主编.环境病理学.第1版.济南:山东科学技术出版社.1990,56-57
[22]DC Purves,IJ Garrod,AD Dayan,et al.A comparison of spongiosis induced in the brain by hexachlorophene,cuprizone and triethyl tin in the Sprague-Dawley rat[J].Human and Experimental Toxicology.1991,10(6):439-444.
[23]N Franco-Pons,M Torrente,MT Colomina,et al.Behavioral deficits in the cuprizone-induced murine model of demyelination/remyelination[J].Toxicol Lett,2007,169(3):205-13.
[24]SW Sun,HF Liang,K Trinkaus,et al.Noninvasive detection of cuprizone induced axonal damage and demyelination in the mouse corpus callosum[J].Magn Reson Med,2006,55(2):302-8.
[25]QZ Wu,Q Yang,HS Cate,et al.MRI identification of the rostral-caudal pattern of pathology within the corpus callosum in the cuprizone mouse model[J].J Magn Reson Imaging.2008,27(3):446-53.
[26]H Jurevics,C Largent,J Hostettler,et al.Alterations in metabolism and gene expression in brain regions during cuprizone-induced demyelination and remyelination[J].J Neurochem.2002,82(1):126-36.
[27]Strang J,Gurling H.computerized tomography and neuropsychological assessment in long-tern high-dose heroin addicts[J].Br J Addict.1989,84:1011-1019.
[28]Campagnoni AT,Macklin WB.Cellular and molecular aspects of myelin protein gene expression[J].Mol Neurobiol.1988,2:41-89.
[29]Roach A,Takahashi N,Pravtcheva D,el at.Chromosomal mapping of mouse myelin basic protein gene and structure and transcription of the partially deleted gene in shiverer mutant mouse[J].Cell.1985,42:149-155.
[30]Newmeyer DD,Ferguson- Miller S.Mitochondria:releasing power for life and unleashing the machineries of death[J].Cell.2003,112(4):481-490.
[31]Thornberry NA,Lazebnik Y.Caspases:enemies within[XJ.Science.1998,281:1312-1316.
[32]Kuida K,Zheng TS,Na S,et al.Decreased apoptosis in the brain and premature lethalitu in CPP32-deficient mice[J].Nature,1996,384(6607):368-372.
[33]Adam JM,Corys.Apoptosomes:engines for caspase activation[J].Curropin Cell Biol.2002,14(6):715.
[34]Cheng EH,Kirsch DQClem PJ,Ravi R,Kastan MB,Bedi A,Ueno K,Hardwick JM.Conversion of Bcl-2 to a Bax-like death effector by caspases[J].Science.1997,278:1966-1968.
[35]Shi Y.Mechanisms of caspase activation and inhibition during apoptosis[J].MolCel.l 2002,9(3):459.
[36]Romero A A,Gross S R,Cheng K Y,et al.An Age-related Increase in Resistance to DNA Damage-induced Apoptotic Cell Death is Associated with Development of DNA Repair Nbchanisms[J].Neurochem,2003,84(6):1275.
[37]Chiang C S,McBride W H,Wither H R.Radiatiorr induced Astrocytic and microglial Responses in Mouse Brain[J].Radiother Oncol,1993,29:60.
[38]W Cammer.The neurotoxicant,cuprizone,retards the differentiation of oligodendrocytes in vitro[J].J Neurol Sci,October 15,1999,168(2):116-120.
[39]Lipton P.Ischemic cell death in brain neurons[J].Physiol Rev,1999,79(4): 1431.
[40]Kurumatani T,Kudo T,Ikura Y,et al.White matter changes in the Gerbil brain under chronic cerebral hypoperfusion[J].Stroke,1998,29:1058-1062.
[41]Jeffrey L.Mason,Arrel Toews,Janell D.Hostettler,et al.Oligodendrocytes and Progenitors Become Progressively Depleted within Chronically Demyelinated Lesions[J].American Journal of Pathology.2004,164:1673-1682.
[42]Herts SW.To Die orNot to Die:an overview of apoptosis and its role in disease[J].JAMA,1998,279:300-307.
[43]Lavrik IN,GolksA,KrammerPH.Caspases:pharmacological manipulation of cell death[J].J Clin Invest,2005,115:2665-2672.