颈交感神经干离断对局灶性脑缺血大鼠脑组织可溶性NSF蛋白附着蛋白受体影响的实验研究
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摘要
目的：在颈交感干离断对局灶性脑缺血大鼠的脑保护作用明确的基础上，探讨其与可溶性NSF蛋白附着蛋白（SNAPs）受体(SNARE)及缺血性脑损伤的关系。
方法：选用Wistar雌性大鼠130只，随机分为大脑中动脉栓塞组（模型组）及大脑中动脉栓塞同时行颈交感干离断组（治疗组），根据栓塞时间又分为6小时、24小时、48小时和72小时四个亚组。使用线栓法制作Wistar大鼠的大脑中动脉闭塞(middle cerebral artery occlusion, MCAO)模型，在此模型基础上，对其行颈交感神经干离断(Transaction of cervicalsympathetic trunk, TCST)术,模拟人类的星状神经节阻滞(satellite ganglionblock，SGB)。观察TCST对实验动物一般状态，神经功能及生存率的影响，应用免疫组织化学染色的方法，观察各个时间点各组大鼠脑组织中海马CA1区神经细胞可溶性NSF蛋白(N-ethylmaleimide-sensitive fusionprotein,NSF)附着蛋白受体（SNAREs）的活性变化。
结果：治疗组动物的一般状态（神经病学评分）及生存率明显优于对照组，同时此组大鼠SNARE的表达明显高于模型组。
结论：颈交感神经干离断可以使SNARE表达明显增加,从而改善缺血性脑卒中动物的一般状态及生存率，对局灶性脑缺血有保护作用。TCST的这些作用可能是通过SNARE介导的途径实现的，为临床上采用星状神经节阻滞治疗缺血性脑卒中提供了有力的实验室依据。
Objective: As known as the transaction of cervical sympathetic trunk canhave the protective effect of focal cerebral ischemia, we explore therelationship with the protective effect of focal cerebral ischemia, the solubleNSF protein attachment protein (SNAPs) receptor (the SNARE) and ischemicbrain injury.
Methods:130Wistar female rats were divided into the middle cerebralartery occlusion group (model group) and the middle cerebral artery occlusionline with the transaction of cervical sympathetic trunk group (treatment group)randomly. According to the embolization time, every groups were divided into6hours,24hours,48hours and72hours of four subgroups. Middle cerebralartery of occlusion (MCAO) model in the brain of Wistar rats were using thesuture method. Based on this model, we make the transaction of cervical ofsympathetic trunk (TCST) to simulate the human satellite ganglion block(SGB). Observe the impact of general status and nerve function of theexperimental rats with TCST. Applicate the immunohistochemical stainingmethod to observe soluble N-ethylmaleimide-sensitive fusion protein (NSF)protein attachment protein receptors (the SNARE) activity in the hippocampalCA1region in each group at each time point in rat brain tissue cells
Results: The general state (neurological score) of the treatment group andsurvival was significantly better than the control group, while rats SNAREexpression was significantly higher than the model group.
Conclusion: The transaction of cervical sympathetic trunk make that theSNARE expression was significantly increased, thereby improving the generalstatus and the survival rate of ischemic stroke in animals, have a protective effect on focal cerebral ischemia. These effects of TCST may be throughSNARE-mediated pathway, and provide a strong experimental basis for theclinical use of stellate ganglion block in treating ischemic stroke.
方法：选用Wistar雌性大鼠130只，随机分为大脑中动脉栓塞组（模型组）及大脑中动脉栓塞同时行颈交感干离断组（治疗组），根据栓塞时间又分为6小时、24小时、48小时和72小时四个亚组。使用线栓法制作Wistar大鼠的大脑中动脉闭塞(middle cerebral artery occlusion, MCAO)模型，在此模型基础上，对其行颈交感神经干离断(Transaction of cervicalsympathetic trunk, TCST)术,模拟人类的星状神经节阻滞(satellite ganglionblock，SGB)。观察TCST对实验动物一般状态，神经功能及生存率的影响，应用免疫组织化学染色的方法，观察各个时间点各组大鼠脑组织中海马CA1区神经细胞可溶性NSF蛋白(N-ethylmaleimide-sensitive fusionprotein,NSF)附着蛋白受体（SNAREs）的活性变化。
结果：治疗组动物的一般状态（神经病学评分）及生存率明显优于对照组，同时此组大鼠SNARE的表达明显高于模型组。
结论：颈交感神经干离断可以使SNARE表达明显增加,从而改善缺血性脑卒中动物的一般状态及生存率，对局灶性脑缺血有保护作用。TCST的这些作用可能是通过SNARE介导的途径实现的，为临床上采用星状神经节阻滞治疗缺血性脑卒中提供了有力的实验室依据。
Objective: As known as the transaction of cervical sympathetic trunk canhave the protective effect of focal cerebral ischemia, we explore therelationship with the protective effect of focal cerebral ischemia, the solubleNSF protein attachment protein (SNAPs) receptor (the SNARE) and ischemicbrain injury.
Methods:130Wistar female rats were divided into the middle cerebralartery occlusion group (model group) and the middle cerebral artery occlusionline with the transaction of cervical sympathetic trunk group (treatment group)randomly. According to the embolization time, every groups were divided into6hours,24hours,48hours and72hours of four subgroups. Middle cerebralartery of occlusion (MCAO) model in the brain of Wistar rats were using thesuture method. Based on this model, we make the transaction of cervical ofsympathetic trunk (TCST) to simulate the human satellite ganglion block(SGB). Observe the impact of general status and nerve function of theexperimental rats with TCST. Applicate the immunohistochemical stainingmethod to observe soluble N-ethylmaleimide-sensitive fusion protein (NSF)protein attachment protein receptors (the SNARE) activity in the hippocampalCA1region in each group at each time point in rat brain tissue cells
Results: The general state (neurological score) of the treatment group andsurvival was significantly better than the control group, while rats SNAREexpression was significantly higher than the model group.
Conclusion: The transaction of cervical sympathetic trunk make that theSNARE expression was significantly increased, thereby improving the generalstatus and the survival rate of ischemic stroke in animals, have a protective effect on focal cerebral ischemia. These effects of TCST may be throughSNARE-mediated pathway, and provide a strong experimental basis for theclinical use of stellate ganglion block in treating ischemic stroke.
引文
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[2]刘冬生，邵燕，欧阳菊香，廖珊.脑梗死预后影响因素的累积logit模型分析[J].中国卫生统计.2010，27（4）:381-382.
[3]吴兆苏,姚崇华,赵冬.我国人群脑卒中发病率、死亡率的流行病学研究[J].中华流行病学杂志，2003，24(3):236-239.
[4]中华医学会.第八届疼痛学分会年会会议论文集[C].成都：[出版者不详]，2011.
[5] Chang-Ki Kang.Ph.D，Seung-Taek Oh B.Sc，Rack Kyung Chung M.D.，Ph.D. Effect of Stellate Ganglion Block on the Cerebrovascular System[J].Anesthesiology2010,113:936–44.
[6]李兴志.颈交感干离断对局灶性脑缺血大鼠生存率影响的实验研究[J].中风与神经疾病杂志，2006，23（4）:436-438
[7]栗志兵,牛丽娟,牛爱清,秦建斌,梁志红.2420例20000次星状神经节阻滞的临床总结[J].实用疼痛学杂志，2009，5（4）：260-264.
[8]王堰婷，陆佩华，盛祖杭.神经递质释放过程中的可溶性NSF附着蛋白受体(SNARE)和相关蛋白[J].生物化学与生物物理进展，2002，29（1）:35-38.
[9]熊良志.星状神经节阻滞对全脑缺血再灌注后家兔海马CA-1区Bax及Bcl-2表达的影响[J].中国临床康复，2006，10（14）:105-107.
[10]蒋燕.星状神经节阻滞的临床研究进展[J].河北医药，2008，30（12）:1969-1970.
[11]刘小立.星状神经节阻滞的方法与用药[J].实用疼痛学杂志，2008，4（2），85.
[12]Longa E Z，Weinstein P R，Carlson S，et al.Reversible middle cerebralartery occlusion without craniectomy in rats[J]. Stroke，1989，20(1):84-91.
[13]Okuda Y, Kitajima T. Influence of stellate ganglion block andelectrical stimulation of the stellate ganglion on bilateral brachialarterial blood flow--is stellate ganglion block effective either unilaterally or bilaterally?[J].Masui,1996,45(12):1465-8.
[14]秦振龙,魏淑明.腰交感神经节切除对大鼠坐骨神经损伤再生的影响
[D].河北:河北医科大学，2005.
[15]郑玉珍，柳顺锁.腰交感神经阻滞对股骨头缺血性坏死修复的影响[D].河北:河北医科大学，2005.
[16]小川秀道.星状神经节阻滞的继发（中枢）性作用：家兔胃溃疡模型的实验研究[J].疼痛学杂志，1991，85-87.
[17]赵广栩.颈交感干离断对浸水大鼠胃粘膜血流量的影响[J].临床麻醉学杂志，2005，21（11）:235-237.
[18]岩间，裕.鼠颈交感干离断与促性腺激素的关系[J].疼痛学杂志，1995，3（1）:6-7.
[19]Uchida K，Tateda T，Hino H. Novel mechanism of action hyp othesizedfor stellate ganglion block related to melatonin[J]．Med Hypotheses，2002，59(4)：446-449．
[20]王贤裕.星状神经节阻滞对兔血浆去甲肾上腺素浓度的影响[J].中华麻醉学杂志，2001,7.365-366.
[21]向勇.星状神经节阻滞对兔炎性疼痛反应和间脑Fos表达的影响[J].医学研究生学报，2005，18（8）:692-695.
[22]熊良志.星状神经节阻滞对兔脑缺血再灌注期血清白细胞介素8的影响[J].中国临床康复，2005，9（10）:98-99.
[23]熊良志.颈交感神经干离断对局灶性脑缺血再灌注后大鼠海马iNOS表达的影响[J].卒中与神经疾病，2008，15（3）:142-147.
[24] Yui Terakawa,DDS，Tatsuya Ichinohe，DDS.PhD，and Yuzuru KanekoDDS PhD. Redistribution of Tissue Blood Flow After Stellate GanglionBlock in the Rabbit[J].Regional Anesthesia and Pain Medicine,34(6)：553-556.
[25]李兴志.颈交感干离断对局灶性脑缺血大鼠血浆中降钙素基因相关肽的影响[J].中国康复医学杂志，2008，23（3）：156-160.
[26]董莹.颈交感干离断对局灶性脑缺血大鼠脑组织病理学及HSP70的影响[D].长春：吉林大学，2007.
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