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.