【作者】 陈玉静； 李林； 李宗信； 陈文强； 黄小波；

【Author】 CHEN Yu-jing LI Lin LI Zong-xin CHEN Wen-qiang HUANG Xiao-bo (Xuanwu Hospital,Capital Medical University,Beijing 100053,China)

【机构】 首都医科大学宣武医院；

【摘要】 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损伤,以及脑康Ⅱ号的保护作用。方法:培养新生Wistar大鼠海马神经元,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0.5g/mL,1.0g/mL,2.0g/mL)预保护24h。利用四甲基偶氮唑蓝法检测细胞存活率,Hoechst 33342免疫荧光染色和流式细胞仪检测细胞凋亡,western blot方法检测凋亡相关蛋白Caspase-3、Bcl-2和Bax的表达。结果:与模型组相比,脑康Ⅱ号能明显提高神经元存活率。免疫荧光染色和流式细胞仪检测结果显示:脑康Ⅱ号各剂量组能明显降低MG引起的凋亡,并能上调Bcl-2蛋白,下调Caspase-3和Bax蛋白的表达。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。更多还原

【Abstract】 Objective:To investigate the injury effect of methylglyoxal(MG) on primary cultured hippocampal neurons of new born rats and the protective effect of NaokangⅡ.Methods:Hippocampal neurons were isolated from new born Wistar rats,and the cultured cells were exposed to various concentrations of MG to prepare a cell injury model.The serum containing NaokangⅡ(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 cell morphology of Hoechst 33342 labeling and flow cytometry of Annexin V/PI method.The expression of Caspase-3,Bcl-2 and Bax proteins was measured by western blot analysis.Results:Pre-incubation of NaokangⅡat all three doses with cultured neurons increased the cell viability compared with MG model group.Results from Hoechst staining,flow cytometric analysis and western blot revealed that pre-incubation of NaokangⅡsignificantly reduced the percentage of MG-induced apoptotic neurons by increasing the expression of Bcl-2 and reducing the expression of Caspase-3 and Bax in the hippocampus. Conclusions:NaokangⅡhas a protective effect against injury of hippocampal neurons induced by MG,which may be related to its anti-apoptotic property.更多还原