脑康Ⅱ号对甲基乙二醛诱导的海马神经元凋亡的影响

Effects of Naokang II on apoptosis of hippocampal neurons induced by methylglyoxal

目的:观察甲基乙二醛(MG)对原代培养的新生鼠海马神经元的凋亡损伤以及脑康Ⅱ号的保护作用。方法:培养新生鼠海马神经元,建立MG损伤神经元模型,并给予脑康Ⅱ号含药血清(分别含生药0.5、1.0、2.0g/mL)预保护24h。利用四甲基偶氮唑蓝法检测细胞存活率,Hoechst33342免疫荧光染色和流式细胞仪检测细胞凋亡。结果:采用100μmolMG(细胞存活率接近于50%)作为本次实验的损伤浓度。脑康Ⅱ号小、中、大各剂量与培养的神经元预孵育24h,细胞存活率比模型组均明显提高(P<0.01)。免疫荧光染色和流式细胞仪检测结果显示:与正常对照组相比,模型组细胞凋亡率显著升高(P<0.01),而脑康Ⅱ号各剂量组能明显降低MG引起的凋亡(P<0.05或P<0.01)。结论:脑康Ⅱ号对MG诱导的海马神经元损伤有保护作用,其机制可能与抑制凋亡有关。

Objective：To investigate the effect of Naokang II on apoptosis of hippocampal neurons induced by methylglyoxal （MG）.Methods：Hippocampal neurons were isolated from new born rats,and the cultured cells were exposed to various concentrations of MG to prepare a cell injury model.The serum containing Naokang II（0.5 g/mL,1.0 g/mL,2.0 g/mL）was pre-incubated with neuron cells for 24 h,respectively.Cell viability was assessed by the MTT assay.Cell apoptosis was determined by Hoechst 33342 labeling and flow cytometry of Annexin V/PI method.Results：100μM MG（corresponding to-50%cell survival）was the best concentration to precede with the following experiments.Pre-incubation of Naokang II at all three doses with cultured neurons increased the cell viability compared with MG model group（P0.01）.Results of Hoechst staining and flow cytometric analysis revealed that a significant increase in apoptosis was observed after MG insult,compared with normal control（P0.05）.Pre-incubation of Naokang II significantly reduced the percentage of apoptotic neurons（P0.05 or P0.01）.Conclusions：Naokang II has a protective effect against injury of hippocampal neurons induced by MG,which may be related to its anti-apoptotic property.