大承气汤对多器官功能障碍综合征大鼠小肠平滑肌细胞Bcl-2和Bax表达的影响
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摘要
背景:多器官功能障碍综合征(multiple organ dysfunction syndrome,MODS)指严重创伤(包括休克、重型胰腺炎等)、严重感染、大面积烧伤、外科大手术和病理产科等原发病发生后,在全身炎症反应综合征(systemic inflammation response syndrome,SIRS)的基础上,同时或序贯发生两个或两个以上脏器功能障碍以致衰竭的临床综合征,是目前腹部外科患者死亡的重要原因。胃肠道作为细菌的储菌库,被认为是促发SIRS和MODS的始动器官与靶部位。MODS可引起胃肠运动功能受损,从而导致细菌移位,肠屏障功能受损。胃肠功能状态被认为是判断MODS患者预后的一项重要指标。促进胃肠运动功能的恢复是有效地阻断MODS向多器官衰竭(multiple organ failure,MOF)发展的有效手段。胃肠内神经—ICC—平滑肌细胞网络是胃肠运动的基本单位。细胞凋亡是个体发育过程中由一系列基因控制的细胞主动死亡过程。平滑肌细胞是胃肠运动的执行者,其结构和功能的正常对于胃肠运动有着重要意义。在平滑肌细胞中,原癌基因Bcl-2蛋白家族是重要的细胞凋亡调节因子,在细胞凋亡中相互联系,相互作用调控细胞凋亡。Bcl-2(B cell lymphoma/leukemia 2)蛋白家族包含抑制和促进细胞凋亡两类功能相反的基因,这两种蛋白的代表分别是Bcl-2和Bax(Bcl-2 associated X protein)。国内医务工作者运用中医药下法的代表方剂大承气汤(Dachengqi Decoction DCQD)对MODS进行中西医结合防治,取得较好效果,显示出中医药在MODS防治中的独特优势。近代研究表明,中医药可从多环节、多靶点地发挥作用,能更好的从整体水平对MODS进行干预、控制。因此深入研究MODS发病机理和大承气汤的治疗机制,对MODS的防治具有重要的临床意义。
目的:本实验以细菌性腹膜炎致MODS大鼠模型为研究对象,观察MODS大鼠模型小肠肌层中的Bcl-2、Bax的分布和平滑肌细胞结构的改变,以及DCQD对MODS大鼠小肠肌层中Bcl-2和Bax的影响。探讨MODS状态下胃肠运动障碍的发生机制和大承气汤防治MODS的机理。
方法:健康成年清洁级Wistar大鼠100只,体重250g左右,雌雄各半,随机分为对照组(20只)、MODS模型组(40只)和MODS+DCQD治疗组(40只)。对照组动物每只腹腔注射1ml生理盐水。MODS模型组和MODS+ DCQD治疗组动物每只腹腔注射1ml E.coloi.混悬液,建立细菌性腹膜炎所致的MODS模型。MODS+DCQD治疗组于造模前2天即给予DCQD灌胃,每次1ml/100g,2次/日。造模24小时后无菌剖腹取存活大鼠的上段小肠组织。
部分小肠置于含2.5%戊二醛固定液的培养皿中,将组织切成1mm×2mm小块,经固定,脱水,浸透,包埋,制备半薄切片,光镜定位下制备超薄切片,最后用透射电镜观察平滑肌细胞形态变化,采集图像。
其它小肠用甲醛固定后,石蜡包埋,用免疫组化(Strept Actividin-Biotin- Complex,SABC法)染色Bcl-2及Bax,然后固定封片。观察各组小肠平滑肌细胞中Bcl-2和Bax的表达及分布情况,采集图像,统计分析。
结果:大体标本和对照组相比,MODS组大鼠胃肠极度扩张,胃肠梗阻征象明显。MODS+DCQD治疗组胃肠扩张、梗阻较MODS组明显减轻。
透射电镜观察:MODS组平滑肌细胞核皱缩,胞浆内细胞器数量明显减少,结构出现异常,超微结构受损明显,出现凋亡征象。较之MODS组,DCQD治疗组平滑肌细胞核形态基本正常,胞浆内细胞器数量较多,形态结构较为清楚,超微结构较MODS组损伤明显减轻。
免疫组织化学检测各组大鼠小肠平滑肌细胞胞浆内均可见Bcl-2和Bax表达,其免疫反应沉淀物为棕黄色,呈颗粒状或匀质状。
对照组Bcl-2和Bax的免疫反应沉淀物在纵肌层和环肌层平滑肌细胞中分布均匀,Bax的较Bcl-2阳性细胞表达稍多。
MODS组Bcl-2较对照组表达强度无明显差异(p>0.05),而Bax较对照组表达明显增多(p<0.05)。同组标本Bax的表达较Bcl-2强烈。
MODS+DCQD治疗组Bcl-2的表达强度均比对照组、MODS组明显增多(p<0.05),而Bax的表达强度较MODS组明显减少(p<0.05),与对照组相比无明显差异(p>0.05)。同组标本Bcl-2的表达较Bax强烈。
结论:1细菌性腹膜炎所致MODS组大鼠胃肠极度扩张,梗阻征象明显;大承气汤能够显著改善其胃肠梗阻症状。2 MODS时大鼠小肠平滑肌细胞结构受到破坏,发生凋亡;DCQD的防治可减轻MODS时平滑肌细胞结构受到的损伤。3 MODS时大鼠小肠平滑肌细胞中的促凋亡基因Bax表达增多,使细胞过度凋亡;DCQD的防治可增加MODS状态下平滑肌细胞抑凋亡基因Bcl-2的表达,减少Bax的表达,下调细胞凋亡水平,保护胃肠平滑肌细胞。
Background:Multiple organ dysfunction syndrome (MODS) is a clinical syndrome, which is induced by the primary disease, such as serious injury ( including shock, acute severe pancreatitis, et al), severe infections, extensive burns, surgical and obstetric major operations and oriented from systemic inflammatory response syndrome (SIRS). In MODS there are two or more organs dysfunction simultaneously or successively. It is the main cause of the death of patients with abdominal surgical diseases. The gastrointestinal tract is a reservoir of bacteria and considered to be the trigger and target site of SIRS and MODS. MODS may cause damage to gastrointestinal motility, bacterial translocation and impaired intestinal epithelial barrier function. The condition of gastrointestinal function is also thought to be the important criterion of evaluation the prognosis of critically ill patients. Promoting the recovery of gastrointestinal motility function could effectively prevent the patient from MODS deteriorating to multiple organ failure (MOF). The network of enteric nerve-ICC-smooth muscle is the basic unit of gastrointestinal motility. Cell apoptosis is the capacity of cells to undergo programmed cell death, which is controlled by a series of genes in the ontogeny course. Smooth muscle cells is the executor of the gastrointestinal motility, its normal structure and function has important significance for gastrointestinal motility. For smooth muscle cells, the original cancer albumen Bcl-2(B cell lymphoma/leukemia 2) protein family is important factors to the apoptosis, connecting and interacting with each other in the apoptosis to regulate the apoptosis, with the two opposite function, inhibition and facilitation. The two kinds of proteins are represented in the Bcl-2 and Bax(Bcl-2 associated X protein). Domestically, The Chinese Medicine are used in treatment of MODS leading to better therapeutic effect which displays distinctive predominance. Recent research has discovered that some Chinese Medicine can produce effect on several aspects of pathogenesis of MODS. Thus, it is important to prevention and cure of MODS that we find out more effective Chinese Medicine.
Objective:To observe the distribution of Bcl-2 and Bax and the structure changes of smooth muscle cells in intestinal muscle of rats with MODS, and the effect of Dachengqi Decoction (DCQD) on Bcl-2 and Bax in intestinal muscle with MODS, to explore the mechanism of gastrointestinal motility disorders leaded by MODS, and to investigate the theory of effect of DCQD on MODS.
Methods:One handred Wistar rats of both sexes weighing about 250g were randomly divided into three groups: control group(n=20), MODS group(n=40) and MODS+DCQD treated group(n=40). The model of MODS was established according to previous study. Briefly, one millilitre suspension of 8×10~8cfu/ml of Escherichia coli strain O127 H6, which contained 10%BaSO4, was injected under sterile conditions into the peritoneal cavity of the rats in the MODS and MODS+DCQD treated group. The rats of control group were injected 1ml of normal saline. The rats of DCQD treated group were administrated by gavage with DCQD (twice a day) before the suspension was injected. Twenty-four hours after injection, the proximal small intestine was taken.
After some of the small intestine was put in 2.5% glutaraldehyde fixative, the tissue was cut into 1mm×2mm pieces, fixed, dehydrated, impregnated, embeded, semi-thin section was prepared, located on the light microscope, the thin slice was prepared. The structural changes of smooth muscle cells were observed, the images were collectted for statistical analysis.
Other intestinal was for formaldehyde fixation, paraffin embedding, immunohistochemistry (SABC method) staining Bcl-2 and Bax, then coverslipping. The expression and distribution of Bcl-2 and Bax in the intestinal muscle were observed, The images were collected for statistical analysis.
Results:Anatomical changes The gastrointestinal tracts were extremely dilated in the rats with MODS compared with in control group. Compared with the MODS group, the dilated gastrointestinal tracts was markedly alleviated in MODS+DCQD treated group.
TEM observation Compared with the control group, the ultrastructural features of the smooth muscle cells in the MODS group displayed severe ultrastructural abnormalities. Injury was displayed in both the perinuclear cytoplasm and the smooth muscle cells processes, the nuclei of smooth muscle cells were shrunken, and the plasma membrane bleb was observed, smooth muscle cells appear apoptosis. Compared with the MODS group, the shape of ultrastructural features of smooth muscle cells in the DCQD group were better, the nuclei was nearly normal.
Immunohistochemistry study Bcl-2 and Bax expression in cytoplasm of smooth muscle cells were observed in rats of various group, The characteristic of Bcl-2 and Bax immuno- reactivity was brown sediment.
In control group, the brown sediment was evenly distributed in circular muscularis and longitudinal muscle layer. In MODS group, the expression of Bax was more than the control group (p<0.05), and the expression of Bcl-2 was not obvious different between the control group(p > 0.05). In the MODS+DCQD treated group, the expression of Bcl-2 was more than MODS group and control group(p<0.05), but the expression of Bax was less than the MODS group(p<0.05) and was not obvious different between the control group(p>0.05).
Conclusion:There was extreme expansion of gastrointestinal and obvious signs of intestinal obstruction in MODS rat induced by bacterial peritonitis; DCQD can significantly alleviate the signs of paralytic intestinal obstruction.
The structure of smooth muscle cells of small intestine in rat with MODS was damaged and smooth muscle cells appeared apoptosis. DCQD can protect structure of smooth muscle cells from destructing in rat with MODS.
The expression of Bax was increased greatly in rat with MODS, DCQD could reduce the expression of Bax and increase the expression of Bcl-2 in rat with MODS. DCQD could protect gastrointestinal smooth muscle cells in MODS.
目的:本实验以细菌性腹膜炎致MODS大鼠模型为研究对象,观察MODS大鼠模型小肠肌层中的Bcl-2、Bax的分布和平滑肌细胞结构的改变,以及DCQD对MODS大鼠小肠肌层中Bcl-2和Bax的影响。探讨MODS状态下胃肠运动障碍的发生机制和大承气汤防治MODS的机理。
方法:健康成年清洁级Wistar大鼠100只,体重250g左右,雌雄各半,随机分为对照组(20只)、MODS模型组(40只)和MODS+DCQD治疗组(40只)。对照组动物每只腹腔注射1ml生理盐水。MODS模型组和MODS+ DCQD治疗组动物每只腹腔注射1ml E.coloi.混悬液,建立细菌性腹膜炎所致的MODS模型。MODS+DCQD治疗组于造模前2天即给予DCQD灌胃,每次1ml/100g,2次/日。造模24小时后无菌剖腹取存活大鼠的上段小肠组织。
部分小肠置于含2.5%戊二醛固定液的培养皿中,将组织切成1mm×2mm小块,经固定,脱水,浸透,包埋,制备半薄切片,光镜定位下制备超薄切片,最后用透射电镜观察平滑肌细胞形态变化,采集图像。
其它小肠用甲醛固定后,石蜡包埋,用免疫组化(Strept Actividin-Biotin- Complex,SABC法)染色Bcl-2及Bax,然后固定封片。观察各组小肠平滑肌细胞中Bcl-2和Bax的表达及分布情况,采集图像,统计分析。
结果:大体标本和对照组相比,MODS组大鼠胃肠极度扩张,胃肠梗阻征象明显。MODS+DCQD治疗组胃肠扩张、梗阻较MODS组明显减轻。
透射电镜观察:MODS组平滑肌细胞核皱缩,胞浆内细胞器数量明显减少,结构出现异常,超微结构受损明显,出现凋亡征象。较之MODS组,DCQD治疗组平滑肌细胞核形态基本正常,胞浆内细胞器数量较多,形态结构较为清楚,超微结构较MODS组损伤明显减轻。
免疫组织化学检测各组大鼠小肠平滑肌细胞胞浆内均可见Bcl-2和Bax表达,其免疫反应沉淀物为棕黄色,呈颗粒状或匀质状。
对照组Bcl-2和Bax的免疫反应沉淀物在纵肌层和环肌层平滑肌细胞中分布均匀,Bax的较Bcl-2阳性细胞表达稍多。
MODS组Bcl-2较对照组表达强度无明显差异(p>0.05),而Bax较对照组表达明显增多(p<0.05)。同组标本Bax的表达较Bcl-2强烈。
MODS+DCQD治疗组Bcl-2的表达强度均比对照组、MODS组明显增多(p<0.05),而Bax的表达强度较MODS组明显减少(p<0.05),与对照组相比无明显差异(p>0.05)。同组标本Bcl-2的表达较Bax强烈。
结论:1细菌性腹膜炎所致MODS组大鼠胃肠极度扩张,梗阻征象明显;大承气汤能够显著改善其胃肠梗阻症状。2 MODS时大鼠小肠平滑肌细胞结构受到破坏,发生凋亡;DCQD的防治可减轻MODS时平滑肌细胞结构受到的损伤。3 MODS时大鼠小肠平滑肌细胞中的促凋亡基因Bax表达增多,使细胞过度凋亡;DCQD的防治可增加MODS状态下平滑肌细胞抑凋亡基因Bcl-2的表达,减少Bax的表达,下调细胞凋亡水平,保护胃肠平滑肌细胞。
Background:Multiple organ dysfunction syndrome (MODS) is a clinical syndrome, which is induced by the primary disease, such as serious injury ( including shock, acute severe pancreatitis, et al), severe infections, extensive burns, surgical and obstetric major operations and oriented from systemic inflammatory response syndrome (SIRS). In MODS there are two or more organs dysfunction simultaneously or successively. It is the main cause of the death of patients with abdominal surgical diseases. The gastrointestinal tract is a reservoir of bacteria and considered to be the trigger and target site of SIRS and MODS. MODS may cause damage to gastrointestinal motility, bacterial translocation and impaired intestinal epithelial barrier function. The condition of gastrointestinal function is also thought to be the important criterion of evaluation the prognosis of critically ill patients. Promoting the recovery of gastrointestinal motility function could effectively prevent the patient from MODS deteriorating to multiple organ failure (MOF). The network of enteric nerve-ICC-smooth muscle is the basic unit of gastrointestinal motility. Cell apoptosis is the capacity of cells to undergo programmed cell death, which is controlled by a series of genes in the ontogeny course. Smooth muscle cells is the executor of the gastrointestinal motility, its normal structure and function has important significance for gastrointestinal motility. For smooth muscle cells, the original cancer albumen Bcl-2(B cell lymphoma/leukemia 2) protein family is important factors to the apoptosis, connecting and interacting with each other in the apoptosis to regulate the apoptosis, with the two opposite function, inhibition and facilitation. The two kinds of proteins are represented in the Bcl-2 and Bax(Bcl-2 associated X protein). Domestically, The Chinese Medicine are used in treatment of MODS leading to better therapeutic effect which displays distinctive predominance. Recent research has discovered that some Chinese Medicine can produce effect on several aspects of pathogenesis of MODS. Thus, it is important to prevention and cure of MODS that we find out more effective Chinese Medicine.
Objective:To observe the distribution of Bcl-2 and Bax and the structure changes of smooth muscle cells in intestinal muscle of rats with MODS, and the effect of Dachengqi Decoction (DCQD) on Bcl-2 and Bax in intestinal muscle with MODS, to explore the mechanism of gastrointestinal motility disorders leaded by MODS, and to investigate the theory of effect of DCQD on MODS.
Methods:One handred Wistar rats of both sexes weighing about 250g were randomly divided into three groups: control group(n=20), MODS group(n=40) and MODS+DCQD treated group(n=40). The model of MODS was established according to previous study. Briefly, one millilitre suspension of 8×10~8cfu/ml of Escherichia coli strain O127 H6, which contained 10%BaSO4, was injected under sterile conditions into the peritoneal cavity of the rats in the MODS and MODS+DCQD treated group. The rats of control group were injected 1ml of normal saline. The rats of DCQD treated group were administrated by gavage with DCQD (twice a day) before the suspension was injected. Twenty-four hours after injection, the proximal small intestine was taken.
After some of the small intestine was put in 2.5% glutaraldehyde fixative, the tissue was cut into 1mm×2mm pieces, fixed, dehydrated, impregnated, embeded, semi-thin section was prepared, located on the light microscope, the thin slice was prepared. The structural changes of smooth muscle cells were observed, the images were collectted for statistical analysis.
Other intestinal was for formaldehyde fixation, paraffin embedding, immunohistochemistry (SABC method) staining Bcl-2 and Bax, then coverslipping. The expression and distribution of Bcl-2 and Bax in the intestinal muscle were observed, The images were collected for statistical analysis.
Results:Anatomical changes The gastrointestinal tracts were extremely dilated in the rats with MODS compared with in control group. Compared with the MODS group, the dilated gastrointestinal tracts was markedly alleviated in MODS+DCQD treated group.
TEM observation Compared with the control group, the ultrastructural features of the smooth muscle cells in the MODS group displayed severe ultrastructural abnormalities. Injury was displayed in both the perinuclear cytoplasm and the smooth muscle cells processes, the nuclei of smooth muscle cells were shrunken, and the plasma membrane bleb was observed, smooth muscle cells appear apoptosis. Compared with the MODS group, the shape of ultrastructural features of smooth muscle cells in the DCQD group were better, the nuclei was nearly normal.
Immunohistochemistry study Bcl-2 and Bax expression in cytoplasm of smooth muscle cells were observed in rats of various group, The characteristic of Bcl-2 and Bax immuno- reactivity was brown sediment.
In control group, the brown sediment was evenly distributed in circular muscularis and longitudinal muscle layer. In MODS group, the expression of Bax was more than the control group (p<0.05), and the expression of Bcl-2 was not obvious different between the control group(p > 0.05). In the MODS+DCQD treated group, the expression of Bcl-2 was more than MODS group and control group(p<0.05), but the expression of Bax was less than the MODS group(p<0.05) and was not obvious different between the control group(p>0.05).
Conclusion:There was extreme expansion of gastrointestinal and obvious signs of intestinal obstruction in MODS rat induced by bacterial peritonitis; DCQD can significantly alleviate the signs of paralytic intestinal obstruction.
The structure of smooth muscle cells of small intestine in rat with MODS was damaged and smooth muscle cells appeared apoptosis. DCQD can protect structure of smooth muscle cells from destructing in rat with MODS.
The expression of Bax was increased greatly in rat with MODS, DCQD could reduce the expression of Bax and increase the expression of Bcl-2 in rat with MODS. DCQD could protect gastrointestinal smooth muscle cells in MODS.
引文
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23.王卫东,陈正堂.Bcl-2/Bax比率与细胞“命运”.中国肿瘤生物治疗杂志,2007,14(4):393-396
24.谭清武.MODS防治的中医及中西医结合研究热点.中国中医急症,2007,(16)3:341-344
25.陈海龙,吴咸中,关凤林,等.中医通里攻下法对多器官功能不全综合征时肠道屏障功能保护作用的实验研究.中国中西医结合杂志,2000,20( 2) : 120- 122
26.陈德昌,杨兴易,景炳文,等.大黄对多器官功能障碍综合征治疗作用的临床研究.中国中西医结合急救杂志,2002,9(1):6-8
27.刘勇,齐清会.MODS肠运动障碍机制和大承气冲剂的治疗作用.中国中西医结合外科杂志,2004,10(6):430-432
28.陈德昌,景炳文,杨兴易,严鸣,李文放,许永华.大黄对创伤后危重病脓毒症患者的治疗作用.中华创伤杂志,2003,19(1):17-19
29.王顺明.中医药防治肠道细菌及内毒素易位的研究进展.中华临床医学杂志,2008,9(5):51-53
30.张栋梁.大承气汤动物含药血浆对Cajal间质细胞三磷酸肌醇受体表达的影响.大连医科大学硕士学位论文,2008
31.余成建.大承气汤动物血清、大黄素和厚朴酚对大鼠小肠Cajal间质细胞内IP3R和RyR表达的影响.大连医科大学硕士学位论文,2009
32.刘健,齐清会.番泻甙对小肠平滑肌细胞收缩功能的影响.中国中西医结合外科杂志,2007,13(1):59-61
33.李毅.多器官功能障碍综合征肠神经-Cajal间质细胞-平滑肌网络形态学变化及大承气汤治疗作用.大连医科大学硕士学位论文,2008
1.龚非力,沈关心,李卓娅,等.医学免疫学.上海:科学技术出版社,2000,6
2.李捷萌,陈彦青,刘荣国.线粒体凋亡途径与Bcl-2家族蛋白研究进展.医学综述,2008,14(4):489-490
3.赵卫红,寿好长,闫福岭.细胞凋亡.郑州:河南医科大学出版社,1997,53-56
4.Adrain C,Martin SJ.The mitochondrial apoptosome:a killer unleashed by the cytochrome seas.Trends Biochem Sci,2001,26(6):390-397
5.Boehning D,Patterson RL,Sedaghat L,Glebova NO,Kurosaki T,Snyder SH.Cytochrome binds to inositol (1 , 4 , 5) trisphosphate receptors , amplifying caldum-dependent apoptosis.NatCell Biol,2003,5:1051- 1061
6.Boehning D,van Roswm DB,Patterson RL,et al.A peptide inhibitor of cytochrome/inositol 1,
4,5-trisphosphate receptor binding blocks intrinsic and extrinsic cell death pathways.Proc Natl Acad Scl U S 2005:102:1466-1471
7.Haeberlein SL.Mitochondrial function in apoptotic neuronal cell death.Neurochem Res,2004,29(3):521-530
8.Reed JC.Apoptosis-based therapies.Nat Rev Drug Discov,2002,1(2):111-121
9.Ishii HH,Gobe GC,Yoneyama J,et al.Role of p53,apoptosis,and cell proliferation in early stage Epstein-Barr virus positive and negative gastric carcinomas.J Clin Pathol 2004,57(12):1306-1311
10.Lockshin RA.Programmed cell death:history and future of a concept.J Soc Biol,2005,199(3):169-173
11.Culmsee C,Plesnila N.Targeting Bid to prevent programmed cell death in neurons.Biochem Soc Trans,2006,34(Pt 6):1334-1340
12.Tsujimoto Y.Cell Death regulation by the Bcl-2 protein family in the mitoehondria.J Cell Physiol,2003,195(2):158-167
13.Raisova M,Hossini AM,Eberle J.The Bax/Bcl-2 ratio determines the susceptibility of human melanoma cells to CD95/Fas-mediated apoptosis.J Invest Dermatol,2001,117(2):333-340
14.De Falco M,Laforgia V,Fedele V.Vasoactive intestinal peptide stimulation modulates the expression of Bcl-2 family members in the adrenal gland of the lizard Podarcis sicula.Histochem J,2001,33(11-12):639-645
15.Degterev A,Boyce M,Yuan J.The channel of death. J Cell, Biol,2001,155(5):695-698
16.Shimizu S,Shinohara Y,Tsujimoto Y.Bax and Bcl-xL independently regulate apoptotic changes of yeast mitochondria that require VDAC but not adenine nucleotide translocator.Oncogene,2000,19(38):4309-4318
17.Tsukaharas S,Yamamoto S,Shew TT,et al.Inhalation of low-level formaldehyde increases the bcl-2/bax expression ratio in the hippocampus of immunologically sensitized mice.Neuro immunomodulation,2006,13(2):63-68
18.迟万好,曹明富,朱睦元.茶多酚诱导肿瘤细胞凋亡的作用.杭州师范学院学报(自然科学版),2001,18(4):46-47
19.Hockenbery G,Csordas G,Das S,et al.Mitoch ond rial calcium signalling and cell death:approaches for assessing the role of mitoch on drial Ca2+ uptake in apoptosis.Cell Calcium,2006,40(5-6):553-560
20.Cheng EH,Kirsch DG,Clen RJ,et al.Conversion of Bcl-2 to a Bax like death effector by caspases.Science,1997,278:1966-1968
21.Wang K,Yin XM,Chao DT,et al.BID:a novel BH3 domain-only death agonist.Genes Dev,1996,10(22):2859-2869
22.Zha JP,Harada H,O sipov K,et al.BH3 domain of BAD is required for hetero dimerization,with Bcl-xl and proapoptotic activity.J Biol Chem,1997,272(39):24101-24104
23.Prakasa Babu P,Yoshida Y,Su M,et al.Immunohistochemical expression of Bcl-2,Bax and cytochrome C following focal cerebral ischemia and effect of hypothermia in rat.Neurosci Lett,2000,291(3):196-200
24.Kuwana T,Mackey MR,Perkins G,et al.Bid,Bax,and lipids cooperate to form supramolecular openings in the outer mitochondrial membrane.Cell,2002,111(3):331-342
25.Mikhailov V,Mikhailova M,Pulkrabek DJ,et al.Bcl-2 prevents Bax oligomerization in the mitochondrial outer membrane.J Biol Chem,2001,276 (21):18361-18374
26.Saito M,Korsmeyer SJ,Schlesinger PH.Bax-dependent transport of cytochrome creconstituted in pure liposomes.Nat Cell Biol,2000,2(8):553-555
27.Antonsson B,Conti F,Ciavatta A,et al.Inhibition of Bax channel forming activity by Bcl-2.Science,1997(6):277-370
28.Oltvai ZN,Milliman CL,Korsmeyer SJ.Bcl-2 heterodimerizes in vivo with a conserved Homolog,Bax,that accelerates programmed cell death.Cell,1993,74(4):609-619
29.Rosset T,Dlivier R,Monney L,et al.Bcl-2 prolongs cell survival after Bax-induced release of cytochrome C.Nature,1998,391-396
30.王卫东,陈正堂.Bcl-2/Bax比率与细胞“命运”.中国肿瘤生物治疗杂志,2007,14(4):393-396
2.岳茂兴.腹部外科疾病并发多器官功能障碍综合征代谢特点及分阶段营养支持治疗.中国危重病急救医学.2003,15:29-31
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4.Ukleja A.Altered GI motility in critically ill Patients:current understanding of pathophysiology,clinical impact,and diagnostic approach.Nutr Clin Pract,2010,25(l):16-25
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9.陈哲宇,齐清会.多器官功能不全综合征大鼠结肠运动功能变化和机制的研究.中华实验外科杂志,2004,21(3):341-344
10.何贵金,吴荣,高沁怡,等.103pd诱导犬胆管增殖平滑肌细胞凋亡及对相关基因的影响.世界华人消化杂志,2005,13(14):1721-1724
11.胡森,高飞.中医药防治多脏器功能障碍综合征回顾与展望.中国中西医结合急救杂志2001,8 6):323-325
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14.Khrupkin VI,Alekseev SA.Syndrome of enteric insufficiency in patients with diffuse peritonitis:evaluation of the severity and outcome of the process.Vestn Khir Im I I Grek,2004,163(2):46-49
15.Balzan S,de Almeida Quadros C,de Cleva R,Zilberstein B,Cecconello I,Bacterial translocation:Overview of mechanisms and clinical impact.Journal of Gastroenterology and Hepatology,2007,22;464–471
16.黄启福.病理学.北京:科学出版社,2007(1):28-29
17.Lockshin RA.Programmed cell death:history and future of a concept.J Soc Biol,2005,199(3):169-173
18.Culmsee C,Plesnila N.Targeting Bid to prevent programmed cell death in neurons.Biochem Soc Trans,2006,34(Pt 6):1334-1340
19.Raisova M,Hossini AM,Eberle J.The Bax/Bcl-2 ratio determines the susceptibility of human melanoma cells to CD95/Fas-mediated apoptosis.J Invest Dermatol,2001,117(2):333-340
20.De Falco M,Laforgia V,Fedele V.Vasoactive intestinal peptide stimulation modulates the expression of Bcl-2 family members in the adrenal gland of the lizard Podarcis sicula.Histochem J,2001,33(11-12):639-645
21.Adrain C,Emma M,Creagh,et al.Apoptosis-associated release of Smac /DIABLO from mitochondria requires aetive caspases and is blocked by Bcl-2. The EMBO Journal,2001,20:6627一6636
22.Haeberlein SL.Mitochondrial function in apoptosis neuronal cell death. Neurochem Res,2004,29(3):521-530
23.王卫东,陈正堂.Bcl-2/Bax比率与细胞“命运”.中国肿瘤生物治疗杂志,2007,14(4):393-396
24.谭清武.MODS防治的中医及中西医结合研究热点.中国中医急症,2007,(16)3:341-344
25.陈海龙,吴咸中,关凤林,等.中医通里攻下法对多器官功能不全综合征时肠道屏障功能保护作用的实验研究.中国中西医结合杂志,2000,20( 2) : 120- 122
26.陈德昌,杨兴易,景炳文,等.大黄对多器官功能障碍综合征治疗作用的临床研究.中国中西医结合急救杂志,2002,9(1):6-8
27.刘勇,齐清会.MODS肠运动障碍机制和大承气冲剂的治疗作用.中国中西医结合外科杂志,2004,10(6):430-432
28.陈德昌,景炳文,杨兴易,严鸣,李文放,许永华.大黄对创伤后危重病脓毒症患者的治疗作用.中华创伤杂志,2003,19(1):17-19
29.王顺明.中医药防治肠道细菌及内毒素易位的研究进展.中华临床医学杂志,2008,9(5):51-53
30.张栋梁.大承气汤动物含药血浆对Cajal间质细胞三磷酸肌醇受体表达的影响.大连医科大学硕士学位论文,2008
31.余成建.大承气汤动物血清、大黄素和厚朴酚对大鼠小肠Cajal间质细胞内IP3R和RyR表达的影响.大连医科大学硕士学位论文,2009
32.刘健,齐清会.番泻甙对小肠平滑肌细胞收缩功能的影响.中国中西医结合外科杂志,2007,13(1):59-61
33.李毅.多器官功能障碍综合征肠神经-Cajal间质细胞-平滑肌网络形态学变化及大承气汤治疗作用.大连医科大学硕士学位论文,2008
1.龚非力,沈关心,李卓娅,等.医学免疫学.上海:科学技术出版社,2000,6
2.李捷萌,陈彦青,刘荣国.线粒体凋亡途径与Bcl-2家族蛋白研究进展.医学综述,2008,14(4):489-490
3.赵卫红,寿好长,闫福岭.细胞凋亡.郑州:河南医科大学出版社,1997,53-56
4.Adrain C,Martin SJ.The mitochondrial apoptosome:a killer unleashed by the cytochrome seas.Trends Biochem Sci,2001,26(6):390-397
5.Boehning D,Patterson RL,Sedaghat L,Glebova NO,Kurosaki T,Snyder SH.Cytochrome binds to inositol (1 , 4 , 5) trisphosphate receptors , amplifying caldum-dependent apoptosis.NatCell Biol,2003,5:1051- 1061
6.Boehning D,van Roswm DB,Patterson RL,et al.A peptide inhibitor of cytochrome/inositol 1,
4,5-trisphosphate receptor binding blocks intrinsic and extrinsic cell death pathways.Proc Natl Acad Scl U S 2005:102:1466-1471
7.Haeberlein SL.Mitochondrial function in apoptotic neuronal cell death.Neurochem Res,2004,29(3):521-530
8.Reed JC.Apoptosis-based therapies.Nat Rev Drug Discov,2002,1(2):111-121
9.Ishii HH,Gobe GC,Yoneyama J,et al.Role of p53,apoptosis,and cell proliferation in early stage Epstein-Barr virus positive and negative gastric carcinomas.J Clin Pathol 2004,57(12):1306-1311
10.Lockshin RA.Programmed cell death:history and future of a concept.J Soc Biol,2005,199(3):169-173
11.Culmsee C,Plesnila N.Targeting Bid to prevent programmed cell death in neurons.Biochem Soc Trans,2006,34(Pt 6):1334-1340
12.Tsujimoto Y.Cell Death regulation by the Bcl-2 protein family in the mitoehondria.J Cell Physiol,2003,195(2):158-167
13.Raisova M,Hossini AM,Eberle J.The Bax/Bcl-2 ratio determines the susceptibility of human melanoma cells to CD95/Fas-mediated apoptosis.J Invest Dermatol,2001,117(2):333-340
14.De Falco M,Laforgia V,Fedele V.Vasoactive intestinal peptide stimulation modulates the expression of Bcl-2 family members in the adrenal gland of the lizard Podarcis sicula.Histochem J,2001,33(11-12):639-645
15.Degterev A,Boyce M,Yuan J.The channel of death. J Cell, Biol,2001,155(5):695-698
16.Shimizu S,Shinohara Y,Tsujimoto Y.Bax and Bcl-xL independently regulate apoptotic changes of yeast mitochondria that require VDAC but not adenine nucleotide translocator.Oncogene,2000,19(38):4309-4318
17.Tsukaharas S,Yamamoto S,Shew TT,et al.Inhalation of low-level formaldehyde increases the bcl-2/bax expression ratio in the hippocampus of immunologically sensitized mice.Neuro immunomodulation,2006,13(2):63-68
18.迟万好,曹明富,朱睦元.茶多酚诱导肿瘤细胞凋亡的作用.杭州师范学院学报(自然科学版),2001,18(4):46-47
19.Hockenbery G,Csordas G,Das S,et al.Mitoch ond rial calcium signalling and cell death:approaches for assessing the role of mitoch on drial Ca2+ uptake in apoptosis.Cell Calcium,2006,40(5-6):553-560
20.Cheng EH,Kirsch DG,Clen RJ,et al.Conversion of Bcl-2 to a Bax like death effector by caspases.Science,1997,278:1966-1968
21.Wang K,Yin XM,Chao DT,et al.BID:a novel BH3 domain-only death agonist.Genes Dev,1996,10(22):2859-2869
22.Zha JP,Harada H,O sipov K,et al.BH3 domain of BAD is required for hetero dimerization,with Bcl-xl and proapoptotic activity.J Biol Chem,1997,272(39):24101-24104
23.Prakasa Babu P,Yoshida Y,Su M,et al.Immunohistochemical expression of Bcl-2,Bax and cytochrome C following focal cerebral ischemia and effect of hypothermia in rat.Neurosci Lett,2000,291(3):196-200
24.Kuwana T,Mackey MR,Perkins G,et al.Bid,Bax,and lipids cooperate to form supramolecular openings in the outer mitochondrial membrane.Cell,2002,111(3):331-342
25.Mikhailov V,Mikhailova M,Pulkrabek DJ,et al.Bcl-2 prevents Bax oligomerization in the mitochondrial outer membrane.J Biol Chem,2001,276 (21):18361-18374
26.Saito M,Korsmeyer SJ,Schlesinger PH.Bax-dependent transport of cytochrome creconstituted in pure liposomes.Nat Cell Biol,2000,2(8):553-555
27.Antonsson B,Conti F,Ciavatta A,et al.Inhibition of Bax channel forming activity by Bcl-2.Science,1997(6):277-370
28.Oltvai ZN,Milliman CL,Korsmeyer SJ.Bcl-2 heterodimerizes in vivo with a conserved Homolog,Bax,that accelerates programmed cell death.Cell,1993,74(4):609-619
29.Rosset T,Dlivier R,Monney L,et al.Bcl-2 prolongs cell survival after Bax-induced release of cytochrome C.Nature,1998,391-396
30.王卫东,陈正堂.Bcl-2/Bax比率与细胞“命运”.中国肿瘤生物治疗杂志,2007,14(4):393-396