脑通汤对缺氧／复氧大鼠海马神经元Bcl－2、Bax和Caspase－3表达的影响

摘要

目的：探讨脑通汤对缺氧／复氧（ H／R ）大鼠海马神经元Bcl－2、Bax和半胱氨酸蛋白酶（ Caspase）－3表达的影响及防治海马神经元凋亡的作用机制。方法取培养9d的海马神经元，随机分为正常细胞组、H／R模型组、正常血清组、脑通汤大、中、小剂量含药血清组。除正常细胞组以外，各组均缺氧24 h再复氧2 h造模。采用免疫组化检测海马神经元Bcl－2、Bax蛋白表达；实时荧光定量PCR法检测Bcl－2、Bax、Caspase－3基因的表达。结果免疫组化显示：与正常细胞比较，模型组Bcl－2蛋白表达下降、Bax蛋白表达明显升高、Bcl－2／Bax比值下调（ P＜0．05）。与模型组比较，脑通汤大、中、小剂量含药血清组 Bcl－2蛋白表达及Bcl－2／Bax比值明显增高，Bax蛋白表达明显下降，呈剂量依赖性（ P＜0．05）；但正常血清组上述指标无显著变化（P＞0．05）。实时荧光定量PCR显示：Bcl－2、Bax mRNA趋势同蛋白表达一致，Caspase－3 mRNA趋势同Bax mRNA表达一致。结论脑通汤可通过上调Bcl－2蛋白及基因表达和Bcl－2／Bax比值，抑制Bax及Caspase－3的过度表达，从而抑制H／R海马神经元凋亡。%Objective To investigate the effects of Naotong soup on the expressions of bcl -2,bax and Caspase-3 hypoxia/reoxygen-ation( H/R) in hippocampal of rats .Methods The hippocampal neurons cultured for 9 d were randomly divided into normal cell ,H/R mod-el,normal serum,large,medium and small doses of Naotong soup medicinal serum groups .The expressions of Bcl2-and Bax protein of hipp-ocampal neurons were detected by immunohistochemistry .Realt-ime fluorescent quantitative PCR was used to detect Bcl -2,Bax and Caspase-3 genes expressions .Results Compared with those of normal cell group ,the expressions of Bcl-2 protein and Bcl-2/Bax were decreased ( P<0.05).However,the expression of Bax protein was significantly increased .Compared with those of model group ,the expressions of Bcl-2 pro-tein and Bcl-2/Bax were significantly increased in Naotong soup medicinal serum groups ,and Bax protein was decreased too (P<0.05).How-ever,the normal serum group showed no significant change (P>0.05).Bcl-2 and Bax mRNAs trend was consistent with the expression of the protein,and Caspase-3 mRNA trend expression was consistent with Bax mRNA .Conclusions Naotong soup could suppress H/R neuronal apoptosis through upregulating the expressions of Bcl-2 protein and gene and Bcl-2/Bax,meanwhile,it could inhibit the expressions of Bax and Caspase-3.