中药脑泰方保护脑缺血再灌注损伤中血管内皮细胞凋亡机制探讨
The mechanism study of Chinese traditional medicine mixture on protection of vascular endothelial cell from apoptosis in cerebral ischemia-reperfusion injury
目的 从凋亡信号通路探讨中药保护脑缺血再灌注损伤中血管内皮细胞凋亡机制.方法 将64只清洁级SD大鼠随机分为4组(每组16只):假手术对照组、脑缺血再灌注组、脑缺血预处理组和中药预处理组;再将脑缺血再灌注组、脑缺血预处理组和中药预处理组按不同时相分为1、7、14及21 d的4个亚组,中药预处理组应用中药脑泰方提取物进行干预.取大鼠大脑中静脉血管内皮细胞,分别行Hoechst 33258染色后计数以及DNA"梯状"条带检测内皮细胞凋亡情况,再应用Western-blot分析中药脑泰方对Caspase-3,8,9活化和Bid裂解的影响情况;从凋亡信号通路探讨中药保护脑缺血再灌注损伤中血管内皮细胞凋亡机制.结果 中药脑泰方提取物可抑制脑缺血再灌注损伤中血管内皮细胞凋亡,能明显抑制缺血再灌注损伤所致的Caspase-3,8,9活化,也抑制了Bid裂解成tBid.结论 中药脑泰方提取物从凋亡信号通路抑制了脑缺血再灌注损伤中血管内皮细胞凋亡.
更多Objective To investigate the mechanisms of Chinese traditional medicine mixture protection of vascular endothelial cell from apoptosis in cerebral ischemia-reperfusion injury. Methods Healthy, clean SD rats were randomly divided into 4 groups: Sham opera-tion group (SOG), cerebral ischemia reperfusion injury group (IRG), cerebral ischemia preconditioning group (IPG) and Chinese tradi-tional medicine mixture preconditioning group (CPG). Furthermore, IRG, IPG and NPC were divided into 4 sub-groups: 1 d, 7d, 14d, 21d subgroup, according to the different time point since ischemia-reperfusion took place. And in CPG, Naotai formula extract was used. Cere-bral vascular endothelial cells of rats were removed and Hoechst 33258 staining and the DNA gradient bands were used to detect the apoptosis of these cells. Then the influence of Naotai formula extract on caspase-3, 8 and 9 activation, and Bid lysis was examined by Western-blot, and the mechanisms of Chinese traditional medicine mixture protection of vascular endothelial cell were investigated from apoptosis signal pathway. Result Naotai formula extract can inhibit the apoptosis of endothelial cells in ischemia-reperfusion injury, and it also can inhibit the activation of caspase-3, 8 and 9, thus inhibit the lysis of Bid into tBid. Conclusion Naomi formula extract inhibit the apoptosis of endo-thelial ceils in ischemia-reperfusion injury via apoptosis signal pathway.
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