大黄素对过氧化氢诱导原代大鼠皮层细胞凋亡的保护作用

Neuroprotective Effects of Emodin on Primary Rat Cortical Neurons Apoptosis Induced by Hydrogen Peroxide

目的:应用过氧化氢(H2O2)孵育原代大鼠皮层细胞制作细胞损伤模型,研究大黄素对模型细胞凋亡的影响。方法:H2O2和大黄素对皮层细胞进行处理,MTT和LDH法检测细胞活性,流式细胞术检测细胞内活性氧(ROS)水平,倒置显微镜、Hoechst33258染色观察细胞形态变化,Western blot法检测Bcl-2和Bax的表达。结果:与正常对照组比较,经150μmol/LH2O2处理后皮层细胞活性显著降低,ROS聚集增加,Hoechst33258荧光染色出现明亮的凋亡小体,大黄素预处理能改善其损害情况。Western blot法显示与正常对照组比较,H2O2可致Bcl-2表达显著减弱,Bax表达显著增强,而大黄素预处理可使其恢复到正常水平。结论:大黄素可拮抗过氧化氢诱导的皮层神经细胞凋亡,具有一定的神经细胞保护功能。

Objective ：To establish a nerve cell injury model by incubating primary rat cortical neurons in the presence of hydrogen peroxide （H2O2） to study the effect of emodin on apoptosis of nerve cells. Methods ：Cortical neurons were incubated with H2O2 and emodin. The cell viability was measured by MTT and cell injury was evaluated by lactate dehydrogenase （LDH）. Flow cytometry was used to evaluate the intracellular level of reactive oxygen species （ROS）. the morphology of cortical cells were observed by inverted microscope and Hoechst 33258 staining. The expression of Bcl-2 and Bax were determined by western blot. Results ：The 150 μmol/L H2O2 significantly decreased the viability of cortical cells and increased the level of intracellular ROS compared to control. Hoechest 33258 fluorescent staining showed brightness nucleosomes in apoptosis cells. The injury state was ameliorated by pretreatment of emodin. Moreover,as detected by Western blot analysis,H2O2 down-regulated the expression of Bcl-2 and up-regulation the expression of Bax compared to control. The state could be reversed. by pretreatment of emodin. Conclusion ：Emodin can resist the adverse effects of H2O2 on increasing apoptotic rates,and it has the protective function towards nerve cells.