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  

The influence of Bcl-2 inhibitor on the alleviation action of astragalus injection on apoptosis of hippocamal neurons of rats induced by hypoxia/hypoglycemia and reoxygenation

作　　者：董雅洁[1] 高维娟[2] 钱涛[3] 卢锴锋[1] 朱炎杰[1] 谢亚芹[1] 

机构地区：[1]承德医学院病理生理学教研室,河北承德067000 [2]河北中医学院,石家庄050200 [3]河北省人民医院,石家庄050051

出　　处：《中国比较医学杂志》2016年第11期11-17,共7页Chinese Journal of Comparative Medicine

基　　金：河北省科技支撑计划项目(13272504D);河北省普通高等学校高层次人才科学研究计划项目(GCC2014031);河北省教育厅青年基金项目(Q2012002);承德医学院自然科学研究计划项目(Q201506);河北省高校重点学科建设项目资助(冀教高【2013】4号病理学与病理生理学)

摘　　要：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。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.

关 键 词：黄芪注射液 Bcl-2抑制剂 缺氧缺糖/复氧复糖 海马神经元 细胞凋亡 CASPASE-3 

分 类 号：R332[医药卫生—人体生理学]