电针人中穴对脑梗死大鼠脑动脉血管舒张功能的调节机制研究

To restore the blood supply of brain tissue is one of the key measures for the treatment of cerebral infarction, to diastole ischemia and its surrounding area vascular can alleviate ischemic brain injury and promote recovery of neurological function. Therefore, it has become one of the important strategies for the treatment of cerebral infarction to adjust the vasodilation. Previous studies show that electro-acupuncture at “Renzhong”(GV26) can promote ischemia and the surrounding area’ s blood supply, then its detailed mechanism is unclear.It can regulate ischemic vasodilatation and to promote blood supply of the surrounding area, It can reduce ischemic brain injury and promote the recovery of neurological function. So regulating vasodilatation has become one of the important strategy for the treatment of cerebral infarction. The study found that the NO / cGMP / PKG pathway is an important mechanism to induce vasodilation. This study on the basis of preliminary studies, to observe NO content and the expression and activity of signal transduction regulatory protein (sGC, cGMP, PKG, of MLCP, HSP20， VASP), the openness and expression of ion channels (VOCC, ROCC,Na / Ca exchange channels open and the expression of IP3-calcium channel), and Ca2 +-ATP enzyme activity. Dynamic observation cerebral infarction after three key factors in the promotion of vascular smooth muscle relaxation sub-signaling pathway variation and acupuncture intervention effect, from the point of view to promote vascular smooth muscle relaxation depth to reveal the molecular mechanism of the acupuncture treatment of cerebral infarction.

恢复脑组织血液供应是治疗脑梗死的关键措施之一，调节缺血及周边区血管舒张可增加血供，减轻脑损伤，促进神经功能恢复。因此调节血管舒张已成为治疗脑梗死的重要策略之一。前期研究显示，电针人中穴可促进缺血及周边区侧枝循环而增加血供，然其深入机制不清。研究发现NO/cGMP/PKG信号通路是诱导血管平滑肌（VSM）舒张的重要机制。本研究拟在前期研究基础上，以此为切入点，以其中诱导VSM舒张的3种途径为主线，以其关键的VSM细胞内NO含量和Ca2+浓度的变化以及信号转导调节蛋白（sGC、cGMP、PKG、MLCP、HSP20、VASP）的表达和活性变化、离子通道（VOCC、ROCC、Na/Ca交换通道、IP3-钙通道）的开放和表达及Ca2+ -ATP酶的活性变化为观测指标，观测脑梗死后该通路促进VSM舒张的信号途径上关键因子的变化规律及针刺干预效应，从促进VSM舒张的角度深入揭示针刺治疗脑梗死的分子机制

本课题通过研究脑梗死发生后脑动脉血管平滑肌细胞中与血管舒张运动相关的NO/cGMP/PKG信号通路中关键的信号转导调节因子NO含量和Ca2+浓度的变化以及信号转导调节蛋白（sGC、cGMP、PKG、MLCP、HSP20、VASP）的表达和活性变化、离子通道（VOCC、ROCC、Na/Ca交换通道、IP3-钙通道）的开放和表达及Ca2+ -ATP酶的活性变化为观测指标，观测脑梗死后该通路促进脑动脉血管平滑肌舒张的信号途径上关键因子的变化规律及针刺干预效应，从促进脑动脉血管平滑肌舒张的角度深入揭示针刺治疗脑梗死的分子机制。.选用Wistar大鼠，随机分为正常组、假手术组、模型组和针刺组。采用Longa线栓法复制脑梗死大鼠模型，以脑表面血管为研究对象，采用免疫印迹法检测sGC、PKG、MLCP、VASP、HSP20、cGMP、Ca2+-ATP酶的蛋白表达，采用RT-PCR法检测VOCC、ROCC、Na/Ca通道、IP3-钙通道的表达量，采用硝酸还原酶法检测NO的含量，采用酶联免疫吸附法检测cGMP的含量。.结果发现大鼠脑梗死后24h内：（1）NO含量、sGC表达量、cGMP表达量随脑梗死时间延长呈逐渐下降趋势，其中NO、cGMP含量下降趋势明显，针刺干预后3h、6h两个时项，针刺效应可抑制NO、cGMP含量的下降趋势，具有一定的时效性；但针刺干预后12h、24h两个时项，cGMP含量下降趋势较NO含量的下降趋势更明显。sGC的下降仅在3h时项有统计学趋势，且在这种趋势可以被针刺的干预效应抑制。（2）VOCC、ROCC、IP3-Ca通道三个亚基表达量变化一致，均随脑梗死时间延长呈整体升高趋势，针刺干预均能抑制其升高趋势，其针刺抑制效应整体以3h、6h两个时项最为明显；Na/Ca交换通道以及Ca2+-ATP酶均随脑梗死时间延长呈整体下降趋势，针刺干预均能抑制其下降趋势，其针刺抑制效应整体以3h、6h两个时项最为明显。（3） VASP、PKG表达量均随脑梗死时间延长呈逐渐下降趋势，其下降趋势可被针刺干预抑制；MLCP、HSP20表达量不受脑梗死时间的影响。. 到目前为止，本课题培养博士、硕士研究生各1名，发表文章2篇。