脑神康胶囊对缺氧复氧损伤大鼠海马神经元凋亡的影响

Effect of Naoshenkang Capsule on neuronal apoptosis and expression of Bcl-2,Bax mRNA in cultured rat hippocampal neurons with anoxia-reoxygenation injury

目的观察脑神康胶囊对缺氧复氧损伤大鼠海马神经元凋亡及其Bcl-2、Bax mRNA表达的影响。方法随机将原代培养大鼠海马神经元分为对照组、损伤组、中药低、中、高浓度组。对照组正常培养,损伤组及中药组建立缺氧复氧损伤模型,中药各浓度组于复氧时换以不同浓度含药血清的培养液,采用MTT法测各组细胞存活率,采用流式细胞仪检测凋亡率,采用RT-PCR法检测Bcl-2和Bax mRNA的表达并计算两者比值。结果损伤组细胞存活率及Bcl-2 mRNA表达明显下降(P均<0.01),凋亡率及Bax mRNA表达显著升高(P均<0.01),Bcl-2/Bax降低(P<0.01);中药各浓度组较损伤组细胞存活率及Bcl-2 mRNA表达均升高(P均<0.01),凋亡率及Bax mRNA表达降低(P<0.01,P<0.05),Bcl-2/Bax增加(P<0.01),且呈明显的剂量依赖性。结论脑神康胶囊对海马神经元缺氧复氧损伤有明显的保护作用,其作用机制可能与其调节凋亡相关蛋白、减少凋亡密切相关。

Objective To explore the effect of Naoshenkang Capsule, a traditional Chinese drug with the function of invigorating Qi and tonifying kidneys on neuronal apoptosis and expression of Bcl-2, Bax mRNA in cultured rat hippocampal neurons with an- oxia-reoxygenation injury. Methods Primarily cultured neonate rat hippocampal neurons were randomly divided into five groups： the control group, the anoxia-reoxygenation group and the small, middle, and large dosage of Chinese Medicine-protective groups. The Chinese Medicine-protective groups were treated with different concentrations of Naoshenkang containing sermn be- fore reoxygenation. The survival rate and apoptosis rate of neurons were determined by MTT and flow cytometry. RT-PCR was used to evaluate mRNA expressions of Bcl-2 and Bax genes. Results Survival rate of cells, Bcl-2 mRNA expression and Bcl-2/ Bax in the anoxia-reoxygenation group were significantly lower （ all P 〈 0.01 ）, but apoptosis rate and Bax mRNA expression were higher （all P 〈 0.01） than those in the control group. Survival rate of cells, Bcl-2 mRNA expression and Bcl-2/Bax in all Chinese Medicine-protective groups were obviously higher （all P 〈 0.01）, but apoptosis rate and Bax mRNA expression were lower （P 〈 0.01, P 〈 0.05） than those in the anoxia-reoxygenation group and demonstrated obvious dosage-effective relationship. Conclusion Naoshenkang Capsule has a protective effect on hippocampal neurons against anoxia-reoxygenation injury, and its mechanism may be that the Chinese medicine can regulate protein expressions of Bcl-2 and Bax and inhibits apoptosis.