Bcl－2抑制剂对黄芪注射液减轻缺氧缺糖／复氧复糖大鼠海马神经元凋亡的影响

摘要

Objective To observe the influence of Bcl-2 inhibitor (TW-37) on the alleviation action of astragalus injection (AI) on apoptosis of hippocamal neurons of rats induced by hypoxia /hypoglycemia and reoxygenation (H/R). Methods The hippocampal neurons were isolated from fetal rat and cultured for eight days .The cells were randomly divided into six groups:normal control group, H/R group, AI group, AI solution group (sterile deionized water), TW-37 group, TW-37 with AI group.All groups were tested after 0 h, 0.5 h, 2 h, 6 h, 24 h, 72 h and 120 h since they were treated with hypoglycemia and reoxygenation after deprived of oxygen and glucose for 30 min except normal control group . The activity of hippocampal neurons were detected with MTT ,The expression level of Caspase-3 was measured by western blotting, the expression of mRNA of Caspase-3 was tested by RT-PCR.Results Compared with normal control group , aside from 0 h group, the activity of neurons decreased obviously ( P <0.05 ), moreover, the protein and mRNA expressions of Caspase-3 were enhanced significantly (P <0.05) in H/R groups.Compared with H/R group, the activity of neurons increased obviously in astragalus group ( P <0.05 ) , and the expressions of Caspase-3 protein and the expression of Caspase-3 mRNA in the AI groups decreased significantly ( P<0.05) besides 0 h group; but there was no significant difference in both AI solution group、TW-37+H/R and TW-37+AI+H/R group ( P>0.05 ) .Conclusions Astragalus injection to increase the neurons activity and decrease the expression level of Caspase -3 in hippocampal neurons of rat by activating the effect of anti-apoptosis of Bcl-2.Bcl-2 is one of the targets exerting the inhibitory action of astragalus injection on apoptosis of hippocamal neurons of rats induced by hypoxia /hypoglycemia and reoxygenation .%目的：探讨黄芪注射液（AI）对Bcl－2抑制剂（TW－37）作用下的缺氧缺糖／复氧复糖（H／R）大鼠海马神经元凋亡的影响。方法取原代培养8 d的胎鼠海马神经元，随机分为6组：正常对照组、H／R组、AI组、AI溶剂对照组、Bcl－2抑制剂（TW－37）＋H／R组、TW－37＋AI＋H／R组。除正常对照组外，各组均进行缺氧缺糖0．5 h，再于复氧复糖后七个时间点（0 h、0．5 h、2 h、6 h、24 h、72 h、120 h）分别进行指标检测。采用MTT法检测细胞活性， western blot法和RT－PCR法检测海马神经元Caspase－3蛋白和mRNA的表达。结果与正常对照组比，除0 h外， H／R组海马神经元活性明显降低（P＜0．05），Caspase－3蛋白及Caspase－3 mRNA表达均明显增强（P＜0．05）；与H／R组相比，除0h外，AI组海马神经元活性明显增高（P＜0．05），Caspase－3蛋白及mRNA表达均明显降低（P＜0．05）；而AI溶剂对照组、TW－37＋H／R组及TW－37＋AI＋H／R组则无显著差异（ P＞0．05）。结论黄芪注射液可通过激活Bcl－2抗凋亡机制增强H／R大鼠海马神经细胞活性、降低Caspase－3表达，Bcl－2是黄芪注射液抑制H／R大鼠海马神经元凋亡的重要靶点之一。