在急性缺血性脑卒中(ICS)治疗中，开通闭塞的动脉血管可引起脑缺血组织再灌注现象，同时也可引发血管内皮细胞损伤及血管内凝血，导致急性缺血性脑卒中再发或脑出血。因此，内皮细胞生物功能对急性缺血性脑卒中发生和预后起着重要的作用。课题组在前期实验研究中发现蒙药扎冲十三味丸对治疗大鼠脑梗死疗效显著，我们分析这可能与激活血管内皮细胞信号传导密切相关。因此，本课题在前期工作基础上，通过建立脑缺血再灌注的大鼠模型，用扎冲十三味丸进行干预，设立对照组，采用Western-blot、紫外线分光光度计、双抗体夹心酶联免疫吸附试验、HE染色法、TTC染色法、电镜下观察法等技术从分子、器官、整体水平进行检测后探讨扎冲十三味丸在大鼠脑缺血再灌注模型中对血管内皮细胞保护作用的机制。

The major purpose in the therapy of acute ischemic stroke (ICS) is to eliminate blood clots and to restore occluded arteries blood flow as soon as possible. However,the recanalization can cause ischemia- reperfusion injury, including vascular endothelial cell injury, inducing intravascular coagulation, and leading to the occurrence of cerebral ischemic stroke. Therefore, it is extremely important to protect the vascular endothelial cells after ischemia-reperfusion in the aspects of structure and function and promote its regeneration which plays an important role in the occurrence and prognosis. Zhachong Shisan Wei Pill performs significant curative effect for the ICS treatment and has many years of clinical and experimental research. In previous research, we found the Zhachong Shisan Wei Pill can reduce ischemia-injury of hypoxia and the effect of nerve cell apoptosis of the rats.Based on the aforementioned researches, we would like to establish the rat model of cerebral ischemia-reperfusion injury. We performed intervention with Zhachong Shisan Wei Pill.Control groups will be set up. We plan to use Western-blot、Ultraviolet Spectrophotometer、 ELISA、Hematoxylin-eosin staining、TTC staining、method of election microscope observational and other advanced techonologies. To discussing the protection mechanism of Mongolian medicine Zhachong Shisan Wei Pill with reperfusion injury of vascular endothelial cell in rats.

目的：验证蒙药扎冲十三味丸对脑缺血再灌注损伤模型大鼠的血管内皮细胞及神经细胞具有保护作用，探讨其保护作用机制。方法:对SD大鼠制备MCAO/R模型，经缺血2h再灌注24h后腹主动脉取血处死大鼠并快速取脑，对脑组织进行TTC染色、HE染色等组织学观察试验。取血清和匀浆后脑组织上清液，对其进行双抗体夹心酶联免疫吸附试验和蛋白印迹实验。最后统计实验数据，对试验进行分析讨论。结果：HE、TTC染色结果显示扎冲十三味丸组大鼠神经损伤程度及梗死面积明显小于模型组。扎冲十三味丸组SOD、GSH-PX、CAT表达量下降程度和MDA表达升高程度不及模型组显著（P<0.05）；与模型组大鼠比较扎冲十三味丸组VEGF表达量升高（P<0.05），NOS和NO表达量降低（P<0.05）。在蛋白印记实验中，与模型组相比较扎冲十三味丸组VEGFR-２表达及磷酸化的Akt表达增强；而模型组脑组织内AQP4的表达量显著高于其他各组。结论：蒙药冲十三味丸对大鼠脑缺血再灌注损伤模型大鼠血管内皮细胞和神经细胞具有的保护作用。其机制为扎冲十三味丸通过提高脑组织SOD、GSH-PX、CAT酶活性，减轻MDA等氧自由基堆积；促进VEGF／VEGFR-２信号通路诱导内皮细胞增值、分化，推动侧支循环的建立；抑制AQP4表达，破坏血管内皮细胞、血脑屏障，形成血管源性脑水肿；促进磷酸化的Akt表达水平，抗神经细胞及血管内皮细胞凋亡；抑制NOS激活并过度生成NO，对神经细胞及血管内皮细胞产生毒性等等多种机制，发挥其对血管内皮细胞和神经细胞的保护作用。