缺血性中风的病因病机及针刺防治机理的实验研究

【作者】 王世军；

【导师】 孙桐；

【作者基本信息】 南京中医药大学 ， 中医基础理论， 2005， 博士

【摘要】 本文通过研究“中风”的病名沿革、病因病机和针灸治疗记载，探讨了祖国医学对中风的认识、针灸治疗中风的历史沿革及其作用机制。 实验研究采用线栓法阻断大脑中动脉(MCAO)复制大鼠局灶性脑缺血模型，在恢复期选用人中内关、曲池足三里、地机经渠三组穴位施以针刺。观察针刺过程中大鼠神经体征、一般状况的改变；检测针刺后血液中一氧化氮(NO)、内皮素(ET)、超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-PX)的变化；并观察脑组织细胞形态学的改变；采用原位杂交技术，检测脑微血管内皮细胞内皮型一氧化氮合酶(ecNOS)及葡萄糖转运蛋白1(Glut-1)、P糖蛋白(P-GP)mRNA的表达。为了比较急性期与恢复期针刺的疗效，我们观察了不同时机电针对MCAO大鼠神经体征的促恢复作用，并检测血液中一氧化氮(NO)、内皮素(ET)、超氧化物歧化酶(SOD)、丙二醛(MDA)的变化；及其对神经元凋亡相关基因Bcl-2、Bax、Caspase-3、神经营养因子3(NT-3)表达的影响。 结果显示，针刺可促进MCAO大鼠神经体征的恢复，其中曲池足三里、人中内关组的促恢复作用明显。针刺可降低血清中NO、MDA的含量，但对ET、GSH-PX及SOD的水平无明显影响。针刺能降低大鼠海马神经元死亡率，增加微血管数目。针刺对脑缺血后海马微血管内皮细胞ecNOS mRNA的表达无明显影响，但可增强神经元ecNOS mRNA的表达。针刺对Glut-1表达的影响不明显。脑缺血后P糖蛋白(P-GP)在微血管内皮细胞表达非常弱，针刺对其的影响尚待进一步研究。 急性期、恢复期针刺的比较研究结果表明，电针可明显促进神经体征的恢复，急性期电针组比恢复期电针组作用明显。电针可以减低脑缺血后升高的NO水平，逆转脑缺血后血浆中ET升高的趋势；电针能够诱导Bcl-2和抑制Bax的蛋白表达，从而抑制神经细胞的凋亡。针刺可促进NT-3的表达，从而促进组织细胞功能及形态结构的恢复。更多还原

【Abstract】 In this study, the terminology, etiology and pathology of Zhongfeng (stroke) were discussed in the view of Traditional Chinese Medicine (TCM). Acupuncture treatment of Zhongfeng was also discussed in many aspects such as the history of acupuncture treatment, the mechanism of acupuncture treatment.Based on our previous research, which indicated that acupuncture can improve cerebral microcirculation and accelerate the recovery of cerebral function in focal cerebral ischemic rats, and for the understanding of the mechanism of acupuncture on the treatment of cerebral ischemic diseases, we studied the change of cerebral function and the expression of functional correlated genes of endothelial cell in the cerebral microvessels after acupuncture in rat with cerebral ischemia. To produce focal cerebral ischemia, middle cerebral artery occlusion (MCAO) was made by nylon thread embolus. The acupoints, Renzhong & Neiguan, Quchi & Zusanli, Diji & Jingqu were chosen for the treatment in different groups in the recovery stage after MCAO. These three groups are compared with MCAO control group and normal control group. The variety of nerve signs and common situation, nitric oxide (NO), endothelin (ET), super-oxide dismutase (SOD), MDA, glutathione peroxidase (GSH-PX) of blood and the morphological change of cerebral cellular were observed. The expression of nitric oxide synthase (ecNOS), glucose transporter-1 (Glut-1) and P-glucoprotein (P-GP) in microvascular endothelial cell were also observed by hybridization in situ.To compare the effects of acupuncture in different stage, we observed the improvement of nerve signs after electro-acupuncture in the acute and chronic ischemic stage. The change of nitric oxide (NO), endothelin (ET), super-oxide dismutase (SOD), MDA of blood, the expressions of bcl-2, bax, caspase-3 and NT-3, which is close related with neuron apoptosis, were also studied.The results showed acupuncture could accelerate the disappearance of pathological nerve signs during recovery stage. Quchi, Zusanli and Renzhong, Neiguan groups showed more obvious effect than other groups. Acupuncture could decline the amount of NO and MDA, but had no obvious effect on ET, GSH-PX and SOD in the blood. Acupuncture could decrease the rate of neuron death and increase the microvascular quantity in the hippocampus. Acupuncture had no obvious influence on the expression ecNOS mRNA in the endothelial cell in the hippocampus, but it could increase the expression of ecNOS更多还原