硫酸镁联合纳洛酮对大鼠创伤性脑损伤脑保护作用的实验研究
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摘要
前言
随着经济和交通手段的日益发展,颅脑损伤的发生率和因伤致残率、致死率均逐年增加。通过药物治疗对颅脑损伤后可能存在的细胞因子变化进行研究,有助于进一步阐明颅脑损伤的机理,从而指导临床治疗,提高颅脑损伤的治愈率、降低死亡率和致残率。肿瘤坏死因子-α属于促炎性细胞因子,在颅脑损伤后表达明显增加,在颅脑外伤后继发性脑损伤中起着重要作用。近年来,大量研究显示了镁剂和纳洛酮在治疗创伤性脑损伤中具有较为肯定的保护作用,但其机制尚未完全阐明。本课题通过Feeney法自由落体动物模型致伤,并给予硫酸镁和纳洛酮治疗,观察大鼠脑创伤后脑组织TNF-α的浓度及脑水肿的变化,探讨两者的脑保护作用机制及其联合用药作用效果。
材料与方法
Wister雄性大鼠60只,体重270～300g,随机分为致伤对照组、纳洛酮组、硫酸镁组及联合用药组,每组再分伤后6h、12h、24h共3个时间点,各时间点每组动物5只。实验大鼠腹腔注射10%水合氯醛(3ml·kg~(-1))麻醉,于中线右侧旁开约2mm,冠状缝后约2mm处钻一直径约5mm骨窗。使用Feeney自由落体损伤装置致伤。致伤成功后各治疗组立即分别给予腹腔注射纳洛酮1mg·kg~(-1)、12.5%的硫酸镁300 mg·kg~(-1)、12.5%的硫酸镁300 mg·kg~(-1)+纳洛酮1mg·kg~(-1),致伤对照组给予等量生理盐水。分别于伤后6h、12h、24h处死大鼠并完整取脑,部分脑组织立即放入10℃甲醛中,固定后制成腊块标本;部分脑组织于-70℃冰箱冰冻保存。按Elliotl公式计算致伤对照组及各治疗组大鼠挫伤灶边缘脑皮质含水量、采用酶联免疫方法(ELISA)测定TNF-α含量,同时取标本做病理学检查。采用SPSS13.0统计软件进行数据处理,计量资料以均数±标准差((?)±s)表示,组间比较采用重复测量数据的方差分析,两两比较用多重比较t检验,P＜0.05为差异有统计学意义。
结果
光镜下发现脑损伤后存在明显炎症反应,细胞水肿及炎症细胞浸润。各组大鼠脑致伤后6h、12h、24h脑组织含水量比较,明显呈增高趋势(各组三时间点间两两比较,均P＜0.05),以致伤后24h脑组织含水量最高。三治疗组与致伤对照组比较,脑组织含水量均有所降低(单一用药组P＜0.05,联合用药组P＜0.01),以联合用药组最明显,且联合用药组与单一用药比较均有显著差异(P＜0.05)。提示:大鼠颅脑致伤后24小时内,脑水肿呈加重趋势,且硫酸镁联合纳洛酮减轻脑水肿作用最明显。损伤组TNF-α水平在损伤后6h、12h、24h三时间点呈上升趋势(P＜0.05)。各治疗组均能抑制TNF-α上升,大鼠致伤后6h、12h、24h脑组织TNF-α的水平与致伤组对照组比较均有差异(P＜0.05,联合用药组P＜0.01),且联合用药组与单一用药组比较亦有统计学意义(P＜0.05),提示:纳洛酮和硫酸镁均能抑制TNF-α水平,且联合用药较单一用药更有效。
结论
大鼠脑损伤后急性期(24小时内)TNF-α的浓度呈增高趋势,随着TNF-α含量的增高,脑水肿也随之明显加重;早期行硫酸镁和纳洛酮治疗能降低脑组织TNF-α的浓度和减轻脑水肿程度,联合用药效果最明显,表明硫酸镁、纳洛酮均对脑创伤后脑损伤有保护作用,并且联合用药效果最佳。
Preface
The incidence of brain injury and accompanied mutilation and mortality rates have increased year by year with the development of the economy and transportation.The abnormal expression of cytokines in traumatic brain injury after drug treatment was investigated to illuminate the mechanism of cerebral injury,and thus to direct the clinical treatment,which produces the elevated recovery rate and decreased mortality and mutilation rates.The tumor necrosis factor-alpha belongs to pro-inflammatory cytokine,which increases obviously after brain injury and plays an important role in the secondary brain injury,the magnesia mixture and naloxone currently have certain protection for traumatic brain injury demonstrated in many researches,but the mechanism has not been illuminated completely.In this study,the animal model for brain injury by the Feeney method of free falling object was established,and given magnesium sulfate and naloxone.The changing in the concentration of TNF-αand brain edema in rats' brain tissue was observed to investigate their brain protective mechanism and the effect of combined medication.
Material and Methods
Total 60 Wister masculinity rats were included in this study,whose body mass were from 270g to 300g,and randomly divided into four groups:vulnerate control group、naloxone group、magnesium sulfate group and combined medication group. Each group was subdivided into three time points:6h、12h、24h after injury,and there were 5 rats in every time point.Experimental rats were anaesthetized through injecting 10%chloral hydrate(3mg/kg)into abdominal cavities,and a diameter of about 5mm bony window was drilled in every rats in about 2mm to the right of the median line and about 2mm to the post of sutura coronalis.The device of Feeney free falling body was used to injury the rats.After injury,every group was immediately given peritoneal injection naloxone(1mg/kg)、12.5%magnesium sulfate(300mg/kg)、12.5%magnesium sulfate(300mg/kg)+naloxone(1mg/kg),and equivalent normal sodium to the control group respectively.The rats were put into death at the time of 6h、12h、24h respectively after injury,and to get brain tissues.Partial brain tissues were immediately put into 10%formaldehyde and made into wax sample after fixing;and partial brain tissues were reserved in refrigerator with temperature70℃below zero.The moisture in the pallium at the edge of contusion foci of rats was calculated according to Elliotl formula in every therapeutic group and the control group、the contents of TNF-αwere determined through ELISA method,and the sample was obtained for pathologic examination.Statistical analysis:all measurement data were expressed as(?)±s,and statistical significance was evaluated by multiple comparison of test and ANOVA.A P value of 0.05 or less was considered to indicate statistical significance.All statistical processes were completed via SPSS(V13.0).
Results
Under the macroscope,obvious inflammatory reaction、cellular edema and inflammatory cell infiltrating were found after cerebral injury.The moisture of brain tissues at 6h、12h、24h after brain injury in rats was compared,and the increasing trend was obvious(every two time points was compared in the three points,P＜0.05),and the moisture of brain tissue at 24h after injury was highest.Comparing the three therapeutic groups and control group,the moisture of brain tissues were all decreased (medication alone P＜0.05,combined medication P＜0.01),most obviously in combined group,and the combined group was more obvious different than medication alone(P＜0.05),these results indicated that during 24h after brain injury of rats,brain edema was increased trend,and combined magnesium sulfate with naloxone was most obvious in relieving brain edema.TNF-αin the injury group was in an upgrading trend at the time of 6h、12h、24h after injury(P＜0.05).Each therapeutic group could inhibit the increasing of TNF-α,and the level of TNF-αat the time point of 6h、12h、24h after injury of rats compared with control group(P＜0.05),which indicated that magnesium sulfate with naloxone can both inhibit the level of TNF-α,and combined medication was better than medication alone.
Conclusion
In the acute phase of brain injury of rats in 24 h,the concentration of TNF-αwas in an increasing trend.As TNF-αincreased,the brain edema was also aggravated obviously.Magnesium sulfate and naloxone were used early to decrease the concentration of TNF-αand brain edema,combined medication was most obvious, which indicated that magnesium sulfate and naloxone can both protect the brain injury after trauma,and combined medication was better.
随着经济和交通手段的日益发展,颅脑损伤的发生率和因伤致残率、致死率均逐年增加。通过药物治疗对颅脑损伤后可能存在的细胞因子变化进行研究,有助于进一步阐明颅脑损伤的机理,从而指导临床治疗,提高颅脑损伤的治愈率、降低死亡率和致残率。肿瘤坏死因子-α属于促炎性细胞因子,在颅脑损伤后表达明显增加,在颅脑外伤后继发性脑损伤中起着重要作用。近年来,大量研究显示了镁剂和纳洛酮在治疗创伤性脑损伤中具有较为肯定的保护作用,但其机制尚未完全阐明。本课题通过Feeney法自由落体动物模型致伤,并给予硫酸镁和纳洛酮治疗,观察大鼠脑创伤后脑组织TNF-α的浓度及脑水肿的变化,探讨两者的脑保护作用机制及其联合用药作用效果。
材料与方法
Wister雄性大鼠60只,体重270～300g,随机分为致伤对照组、纳洛酮组、硫酸镁组及联合用药组,每组再分伤后6h、12h、24h共3个时间点,各时间点每组动物5只。实验大鼠腹腔注射10%水合氯醛(3ml·kg~(-1))麻醉,于中线右侧旁开约2mm,冠状缝后约2mm处钻一直径约5mm骨窗。使用Feeney自由落体损伤装置致伤。致伤成功后各治疗组立即分别给予腹腔注射纳洛酮1mg·kg~(-1)、12.5%的硫酸镁300 mg·kg~(-1)、12.5%的硫酸镁300 mg·kg~(-1)+纳洛酮1mg·kg~(-1),致伤对照组给予等量生理盐水。分别于伤后6h、12h、24h处死大鼠并完整取脑,部分脑组织立即放入10℃甲醛中,固定后制成腊块标本;部分脑组织于-70℃冰箱冰冻保存。按Elliotl公式计算致伤对照组及各治疗组大鼠挫伤灶边缘脑皮质含水量、采用酶联免疫方法(ELISA)测定TNF-α含量,同时取标本做病理学检查。采用SPSS13.0统计软件进行数据处理,计量资料以均数±标准差((?)±s)表示,组间比较采用重复测量数据的方差分析,两两比较用多重比较t检验,P＜0.05为差异有统计学意义。
结果
光镜下发现脑损伤后存在明显炎症反应,细胞水肿及炎症细胞浸润。各组大鼠脑致伤后6h、12h、24h脑组织含水量比较,明显呈增高趋势(各组三时间点间两两比较,均P＜0.05),以致伤后24h脑组织含水量最高。三治疗组与致伤对照组比较,脑组织含水量均有所降低(单一用药组P＜0.05,联合用药组P＜0.01),以联合用药组最明显,且联合用药组与单一用药比较均有显著差异(P＜0.05)。提示:大鼠颅脑致伤后24小时内,脑水肿呈加重趋势,且硫酸镁联合纳洛酮减轻脑水肿作用最明显。损伤组TNF-α水平在损伤后6h、12h、24h三时间点呈上升趋势(P＜0.05)。各治疗组均能抑制TNF-α上升,大鼠致伤后6h、12h、24h脑组织TNF-α的水平与致伤组对照组比较均有差异(P＜0.05,联合用药组P＜0.01),且联合用药组与单一用药组比较亦有统计学意义(P＜0.05),提示:纳洛酮和硫酸镁均能抑制TNF-α水平,且联合用药较单一用药更有效。
结论
大鼠脑损伤后急性期(24小时内)TNF-α的浓度呈增高趋势,随着TNF-α含量的增高,脑水肿也随之明显加重;早期行硫酸镁和纳洛酮治疗能降低脑组织TNF-α的浓度和减轻脑水肿程度,联合用药效果最明显,表明硫酸镁、纳洛酮均对脑创伤后脑损伤有保护作用,并且联合用药效果最佳。
Preface
The incidence of brain injury and accompanied mutilation and mortality rates have increased year by year with the development of the economy and transportation.The abnormal expression of cytokines in traumatic brain injury after drug treatment was investigated to illuminate the mechanism of cerebral injury,and thus to direct the clinical treatment,which produces the elevated recovery rate and decreased mortality and mutilation rates.The tumor necrosis factor-alpha belongs to pro-inflammatory cytokine,which increases obviously after brain injury and plays an important role in the secondary brain injury,the magnesia mixture and naloxone currently have certain protection for traumatic brain injury demonstrated in many researches,but the mechanism has not been illuminated completely.In this study,the animal model for brain injury by the Feeney method of free falling object was established,and given magnesium sulfate and naloxone.The changing in the concentration of TNF-αand brain edema in rats' brain tissue was observed to investigate their brain protective mechanism and the effect of combined medication.
Material and Methods
Total 60 Wister masculinity rats were included in this study,whose body mass were from 270g to 300g,and randomly divided into four groups:vulnerate control group、naloxone group、magnesium sulfate group and combined medication group. Each group was subdivided into three time points:6h、12h、24h after injury,and there were 5 rats in every time point.Experimental rats were anaesthetized through injecting 10%chloral hydrate(3mg/kg)into abdominal cavities,and a diameter of about 5mm bony window was drilled in every rats in about 2mm to the right of the median line and about 2mm to the post of sutura coronalis.The device of Feeney free falling body was used to injury the rats.After injury,every group was immediately given peritoneal injection naloxone(1mg/kg)、12.5%magnesium sulfate(300mg/kg)、12.5%magnesium sulfate(300mg/kg)+naloxone(1mg/kg),and equivalent normal sodium to the control group respectively.The rats were put into death at the time of 6h、12h、24h respectively after injury,and to get brain tissues.Partial brain tissues were immediately put into 10%formaldehyde and made into wax sample after fixing;and partial brain tissues were reserved in refrigerator with temperature70℃below zero.The moisture in the pallium at the edge of contusion foci of rats was calculated according to Elliotl formula in every therapeutic group and the control group、the contents of TNF-αwere determined through ELISA method,and the sample was obtained for pathologic examination.Statistical analysis:all measurement data were expressed as(?)±s,and statistical significance was evaluated by multiple comparison of test and ANOVA.A P value of 0.05 or less was considered to indicate statistical significance.All statistical processes were completed via SPSS(V13.0).
Results
Under the macroscope,obvious inflammatory reaction、cellular edema and inflammatory cell infiltrating were found after cerebral injury.The moisture of brain tissues at 6h、12h、24h after brain injury in rats was compared,and the increasing trend was obvious(every two time points was compared in the three points,P＜0.05),and the moisture of brain tissue at 24h after injury was highest.Comparing the three therapeutic groups and control group,the moisture of brain tissues were all decreased (medication alone P＜0.05,combined medication P＜0.01),most obviously in combined group,and the combined group was more obvious different than medication alone(P＜0.05),these results indicated that during 24h after brain injury of rats,brain edema was increased trend,and combined magnesium sulfate with naloxone was most obvious in relieving brain edema.TNF-αin the injury group was in an upgrading trend at the time of 6h、12h、24h after injury(P＜0.05).Each therapeutic group could inhibit the increasing of TNF-α,and the level of TNF-αat the time point of 6h、12h、24h after injury of rats compared with control group(P＜0.05),which indicated that magnesium sulfate with naloxone can both inhibit the level of TNF-α,and combined medication was better than medication alone.
Conclusion
In the acute phase of brain injury of rats in 24 h,the concentration of TNF-αwas in an increasing trend.As TNF-αincreased,the brain edema was also aggravated obviously.Magnesium sulfate and naloxone were used early to decrease the concentration of TNF-αand brain edema,combined medication was most obvious, which indicated that magnesium sulfate and naloxone can both protect the brain injury after trauma,and combined medication was better.
引文
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28 樊永中,朱凤清,周幽心.大鼠脑挫裂伤后Fos蛋白表达及硫酸镁的作用.苏州医学院学报,2000:20(7):607-610.
29 石全红,支兴刚,何朝晖.硫酸镁对大鼠创伤性脑损伤后神经细胞凋亡的影响.重庆医科大学学报,2004;29(1):48-50.
30 Heath DL,Vink R.Optimization of magnesium therapy after severe diffuse axonal brain injury in rats.J Pharmacal Exp Ther,1999;288:1311-1316.
31 Deanne L,Heath DL,Vink R.Improved motor outcome in response to magnesium therapy received up to 24 hours after traumatic diffuse axonal brain injury in rats.Neurosurg,1999:90:504-509.
32 张文川,李世亭,徐顺清,等.颅脑外伤急性期血清镁离子浓度的变化.中华急诊医学杂志,2005:14(11):928-930.
33 Hoane MR,Barth TM.The window of opportunity for administration of magnesium therapy following focal brain injury is 24h but is task dependent in the rat.Physiol Behav,2002;76:271-280.
34 Muir KW,Lees KR.Dose optimization of intravenous magnesium sulfatea)after acute stroke.Stroke,1998;29:918-923.
35 Khan OH,Enno T,Del Bigio MR.Magnesium sulfate therapy is of mild benefit to young rats with kaolin-induced hydrocephalus.Pediatr Res,2003;53:970-976.
36 Veyna RS,Seyfried D,Burke DG,et al.Magnesiun sulfate therapy after anenurysmal subarachnoid hemorrhage.J Neurosurg,2002;96:510-514.
37 金尔伦全国多中心双盲临床研究课题组.金尔伦(盐酸钠络酮)治疗急性颅脑损伤患者随机双盲多中心前瞻性临床研究.中华神经外科杂志,2001:17(3):135-139.
38 曾小明,何坚汉,王雪莹,等。金尔伦在急性重症脑外伤的临床应用研究.海军医学杂志,2006;27(2):119-121.
39 江基尧,朱诚,王成海,等.急性颅脑损伤患者脑脊液β 2ELI含量变化及其临床意义.中华医学杂志.1988:68(8):422-424.
40 Tseng EE,Brock MV,Kwon CC,et al.Increased int racerebralexcitatoryamino acids and nitric oxide afterhypot hermit circulatory arrest.Ann Thorac Sury,1999;67:371-376.
41 Yang JC,Shan J,NgKF,et al.Morphine and methadone haveelifferent effect on cakium channelcurrent s in neuroblastoma cells.Brain Res,2000;870:199-203.
42 Przewlocki R,Parsons KL,Sweaney DD,et al.Opioid onhancement NMD Areceptors and L -type claliccmchannels in cultured hippocampalneurons.Neurosu,1999;19:9705-9715.
43 曲丽辉,李玉荣,贾淑伟,等纳洛酮对脑缺血大鼠皮层SEP和Ca2+-ATP酶活性的影响.哈尔滨医科大学学报.2004~38(1):45-47.
44 黎沾良主编.现代危重病学.安徽科学技术出版社,1998:114-115.
45 Chun-Jung Chen,Fu-Chou Cheng,et al.Effects of naloxone on lactate,pyruvate metabolism and antioxidant enzyme activity in rat cerebral isehemia/reperfusion.Neuroscience Letters 2000;287:113-116.
46 Vink R,Mclntosh TK,et al.Opiate antagonist nalmefene improves intracellular free Mg2+,bioenergetic state,and neurologic outcome following traumatic brain injury in rats.Neurosci 1990 Nov;10(11):3524-30.
47 Tymianski M,Tator CH.Normal and abnormal calcium homeostasis in neurons:a basis for the pathophysiology of traumatic and ischemic central nervous system injury.Neurosurg,1996;38:1176-1195.
48 Fade AI,Demediuk P,Panter SS,et al.The role of ecitatory amino acids and NMDA receotors in traumatic head injury.Science,1989,;244:798-800.
49 Liu B,Du L,Kong LY,et al.Reduction by naloxone of lipopolysaccharide-induced neurotoxicity in mouse cortical neuron-glia co-cultures.Neuroscience 2000;97(4):749-56.
50 Shibata S,Tominaga K,Watanabe S.Kappa-Opioid receptor agonist protects against ischemic reduction of 2-deoxyglucose uptake in morphine-tolerant rats.Eur J Pharmacol 1995 Jun 12;279(2-3):197-202.
51 Kugawa F,Arae K,Ueno A,et al.Buprenorphine hydrochloride induces apoptosis in NG108215 nerve cells.Eur J Pharmacol,1998;347105-112.
52 黄志雄,徐安定,徐如祥,等.纳洛酮对大鼠颅脑损伤后神经细胞凋亡的影响.中华实验外科杂志,2002;19(6):581-582.
53 Vink R,Nimmo AJ.Novel therapies in development for the treatment of traumatic brain injury.Expert Opin Investig Drugs,2002;11(10):1375-1386.
54 Martina S,Elizabeth C,Elad I.L,et al.Magnesium infusion for vasospasm prophylaxis after subarachnoid hemorrhage.Neurosurg,2006;105:723-729.
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7 Vink R,O'Connor CA,Nimmo AJ,et al.Magnesium attenuates persistent functional deficits following diffuse traumatic-brain injury in rats.Neurosci Lett.2003;336(1):41-44.
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11 周国胜,赵洪洋,朱贤立,等。大剂量硫酸镁对脑外伤治疗作用的研究。中华创伤杂志,2001:11(3):161-162.
12 Keith W,Muir MRCP,Kennedy R,et al.A randomized double blind Placebo controlled pilot trial of intravenous magnesium sulfate in acutestroke.Stroke,1995;26:1183.
13 Muir KW.New experimental and clinical data on the efficacy of pharmacological magnesium infusion in cerebral infarcts.Magnes Res,1998;11(1):43-56.
14 Vink R,Heath DL,McIntosh TK,et al.Acute and prdonged alterations in brain free magnesium following fluid percussion-inducde brain trauma in rats.Neu rochem,1996;66(6):2477-2483.
15 McDonald JW,Silverstein FS,Johnston MV.Magnesium reduces N-methy-aspartate(NMDA)-mediated brain injury in perinatal rats.Neurosci Lett,1990;109(1-2):234-238.
16 Heath DL.Characterisation of the neuroprotective role of magnesium indiffuse traumatic brain injury.James Cook Univrsity of North Queens land,Townsville,Queensland,Australia,1997.
17 Altura BM,Gebrewold A,Zhang A,et al.Low extracellular magnesium ions induce lipid peroxidation and activation of nuclear factor-kappa B in canine cerebral vascular smooth muscle:possible relation to traumatic brain injury and strokes.Neurosci Lett,2003;341(3):189-192.
18 Bariskaner H,Ustun ME,Yosunkaya A,et al.Effects of magnesium sulfate on tissue lactate and malondialdehyde levels after cerebral ischemia.Pharmacology,2003;68(3):162-168.
19 Ustun ME,Duman A,Ogun CO,et al.Effects of nimodipine and magnesium sulfate on endogenous antioxidant levels in brain tissue after experimental head trauma.J Neurosurg Anesthesiol,2001;13(3):227-232.
20 Ram Z,Sadeh M,Shacked I,et al.Magnesium sulfate reverses experimental delayed cerebral vasospasm after subarachnoid hemorrhage in rats.Stroke,1991;22:922-927.
21 Van der Hel WS,Van den Bergh WM,Nicolay K,et al.Suppression of cortical spreading depressions after magnesium treatment in the rat.Neuroreport,1998;9(10):2179-2182.
22 Vink R,Cernak I.Regulation of intracellular free magnesium in central nervous system injury.Front Biosci,2000;5:656-665.
23 Saatman KE,Bareyre FM,Grady MS,et al.Acute cytoskeletal alterations and cell death induced by experimental brain injury are attenuated by magnesium treatment and exacerbated by magnesium deficiency.J Neuropathol Exp Neurol,2001;60(2):183-194.
24 Krueger RC,Santore MT,Dawson G,et al.Increasde extracellular magnesium modulates proliferation in fetal neural cells in culture.Brain Res Dev Brain Res,2001;127(2):99-109.
25 Rink A,Fung KM,Trojanowski JQ,et al.Evidence ofapoptotic cell death after expeimental traumatic brain injury in the rat.Am J Path1,1995;147(6):1575-1583.
26 张亚卓,王忠诚,郑毓书,等.猫中脑损伤后细胞凋亡及相关基冈表达的意义.中华神经外科杂志,1999;15(4):233-236.
27 Muir J K,Raghupathi R,Emery DL,et al.Postinjury magnesium treatment attenuates traumatic brain injury-induced cortical induction of p53 mRNA in rats Exp Neurol,1999;159(2):584-593.
28 樊永中,朱凤清,周幽心.大鼠脑挫裂伤后Fos蛋白表达及硫酸镁的作用.苏州医学院学报,2000:20(7):607-610.
29 石全红,支兴刚,何朝晖.硫酸镁对大鼠创伤性脑损伤后神经细胞凋亡的影响.重庆医科大学学报,2004;29(1):48-50.
30 Heath DL,Vink R.Optimization of magnesium therapy after severe diffuse axonal brain injury in rats.J Pharmacal Exp Ther,1999;288:1311-1316.
31 Deanne L,Heath DL,Vink R.Improved motor outcome in response to magnesium therapy received up to 24 hours after traumatic diffuse axonal brain injury in rats.Neurosurg,1999:90:504-509.
32 张文川,李世亭,徐顺清,等.颅脑外伤急性期血清镁离子浓度的变化.中华急诊医学杂志,2005:14(11):928-930.
33 Hoane MR,Barth TM.The window of opportunity for administration of magnesium therapy following focal brain injury is 24h but is task dependent in the rat.Physiol Behav,2002;76:271-280.
34 Muir KW,Lees KR.Dose optimization of intravenous magnesium sulfatea)after acute stroke.Stroke,1998;29:918-923.
35 Khan OH,Enno T,Del Bigio MR.Magnesium sulfate therapy is of mild benefit to young rats with kaolin-induced hydrocephalus.Pediatr Res,2003;53:970-976.
36 Veyna RS,Seyfried D,Burke DG,et al.Magnesiun sulfate therapy after anenurysmal subarachnoid hemorrhage.J Neurosurg,2002;96:510-514.
37 金尔伦全国多中心双盲临床研究课题组.金尔伦(盐酸钠络酮)治疗急性颅脑损伤患者随机双盲多中心前瞻性临床研究.中华神经外科杂志,2001:17(3):135-139.
38 曾小明,何坚汉,王雪莹,等。金尔伦在急性重症脑外伤的临床应用研究.海军医学杂志,2006;27(2):119-121.
39 江基尧,朱诚,王成海,等.急性颅脑损伤患者脑脊液β 2ELI含量变化及其临床意义.中华医学杂志.1988:68(8):422-424.
40 Tseng EE,Brock MV,Kwon CC,et al.Increased int racerebralexcitatoryamino acids and nitric oxide afterhypot hermit circulatory arrest.Ann Thorac Sury,1999;67:371-376.
41 Yang JC,Shan J,NgKF,et al.Morphine and methadone haveelifferent effect on cakium channelcurrent s in neuroblastoma cells.Brain Res,2000;870:199-203.
42 Przewlocki R,Parsons KL,Sweaney DD,et al.Opioid onhancement NMD Areceptors and L -type claliccmchannels in cultured hippocampalneurons.Neurosu,1999;19:9705-9715.
43 曲丽辉,李玉荣,贾淑伟,等纳洛酮对脑缺血大鼠皮层SEP和Ca2+-ATP酶活性的影响.哈尔滨医科大学学报.2004~38(1):45-47.
44 黎沾良主编.现代危重病学.安徽科学技术出版社,1998:114-115.
45 Chun-Jung Chen,Fu-Chou Cheng,et al.Effects of naloxone on lactate,pyruvate metabolism and antioxidant enzyme activity in rat cerebral isehemia/reperfusion.Neuroscience Letters 2000;287:113-116.
46 Vink R,Mclntosh TK,et al.Opiate antagonist nalmefene improves intracellular free Mg2+,bioenergetic state,and neurologic outcome following traumatic brain injury in rats.Neurosci 1990 Nov;10(11):3524-30.
47 Tymianski M,Tator CH.Normal and abnormal calcium homeostasis in neurons:a basis for the pathophysiology of traumatic and ischemic central nervous system injury.Neurosurg,1996;38:1176-1195.
48 Fade AI,Demediuk P,Panter SS,et al.The role of ecitatory amino acids and NMDA receotors in traumatic head injury.Science,1989,;244:798-800.
49 Liu B,Du L,Kong LY,et al.Reduction by naloxone of lipopolysaccharide-induced neurotoxicity in mouse cortical neuron-glia co-cultures.Neuroscience 2000;97(4):749-56.
50 Shibata S,Tominaga K,Watanabe S.Kappa-Opioid receptor agonist protects against ischemic reduction of 2-deoxyglucose uptake in morphine-tolerant rats.Eur J Pharmacol 1995 Jun 12;279(2-3):197-202.
51 Kugawa F,Arae K,Ueno A,et al.Buprenorphine hydrochloride induces apoptosis in NG108215 nerve cells.Eur J Pharmacol,1998;347105-112.
52 黄志雄,徐安定,徐如祥,等.纳洛酮对大鼠颅脑损伤后神经细胞凋亡的影响.中华实验外科杂志,2002;19(6):581-582.
53 Vink R,Nimmo AJ.Novel therapies in development for the treatment of traumatic brain injury.Expert Opin Investig Drugs,2002;11(10):1375-1386.
54 Martina S,Elizabeth C,Elad I.L,et al.Magnesium infusion for vasospasm prophylaxis after subarachnoid hemorrhage.Neurosurg,2006;105:723-729.