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目的探讨醒脑静对脑缺血再灌注(CIR)过程中神经细胞自噬的疗效及可能作用机制。方法用栓线法制备90只大脑中动脉栓塞大鼠模型,选取符合要求的模型大鼠随机分为2大组:MRI组(20只)和WB组(70只),MRI组随机分为2小组:模型对照组10只,醒脑静组10只;WB组随机分为7组:模型组,溶媒组,醒脑静组,3-MA组,3-MA+醒脑静组,雷帕霉素(Rapa)组,Rapa+醒脑静组(每组10只)。用磁共振成像检测MRI组大鼠用药前后模型脑梗的体积,计算脑梗体积占全脑体积的比例;用免疫印迹法检测WB组大鼠细胞内自噬相关蛋白的表达。结果 MRI组:各组大鼠给药24h后的脑梗体积比例均大于给药前的脑梗体积比例,差异具有统计学意义(P<0.05);醒脑静组的脑梗体积比例小于模型对照组,差异具有统计学意义(P<0.05)。WB组:与模型组相比,醒脑静组、3-MA组与3-MA+醒脑静组的Beclin1和LC3的表达量均降低,Bcl-2的表达量升高,差异具有统计学意义(P<0.05),且3-MA组+醒脑静组优于3-MA组与醒脑静组(P<0.05);Rapa组与Rapa组+醒脑静组Beclin1和LC3的表达量均高于模型组、醒脑静组、3-MA组与3-MA组+醒脑静组,Bcl-2表达量低于模型组、醒脑静组、3-MA+醒脑静组(P<0.05),且Rapa+醒脑静组Beclin1和LC3低于Rapa组,Bcl-2高于Rapa组(P<0.05)。Bcl-2与Beclin1之间的表达量呈负相关。结论醒脑静可抑制大鼠脑缺血再灌注过程中的自噬反应,提高受损神经细胞的存活率。
Objective To investigate the effect of Xingnaojing on neuronal autophagy during cerebral ischemia-reperfusion (CIR) and its possible mechanism. Methods 90 middle cerebral artery occlusion rat models were prepared by thread embolization. The rats were selected and randomly divided into 2 groups: MRI group (20) and WB group (70). The MRI group was randomly divided into 2 groups Group: 10 in model control group and 10 in Xingnaojing group. WB group were randomly divided into 7 groups: model group, vehicle group, Xingnaojing group, 3-MA group, 3-MA + Xingnaojing group, Rapa group, Rapa + Xingnaojing group (10 in each group). The volume of cerebral infarction in the MRI group was measured by MRI before and after treatment. The proportion of cerebral infarction volume to whole brain volume was calculated. The expression of autophagy-related protein in WB group was detected by Western blotting. Results MRI group: The proportion of cerebral infarction volume in rats in each group was higher than that before administration, the difference was statistically significant (P <0.05); the proportion of brain infarction in Xingnaojing group was smaller than that in model group Control group, the difference was statistically significant (P <0.05). Compared with the model group, the expression of Beclin1 and LC3 in Xingnaojing group, 3-MA group and 3-MA + Xingnaojing group were lower than those in the model group, and the expression of Bcl-2 was significantly increased (P <0.05), and the expression of Beclin1 and LC3 in 3-MA group and Xingnaojing group were better than those in 3-MA group and Xingnaojing group (P <0.05) The expression of Bcl-2 in model group, Xingnaojing group and 3-MA + Xingnaojing group was significantly higher than that in model group, Xingnaojing group, 3-MA group and 3-MA group + Xingnaojing group (P < 0.05). Beclin1 and LC3 in Rapa + Xingnaojing group were lower than those in Rapa group, and Bcl-2 was higher than Rapa group (P <0.05). The expression of Bcl-2 and Beclin1 was negatively correlated. Conclusion Xingnaojing can inhibit the autophagy in rats during cerebral ischemia reperfusion and improve the survival rate of damaged nerve cells.