Effects of Senegenin against Hypoxia/Reoxygenation-Induced Injury in PC12 Cells  

作　　者：朱小青 李学敏 赵严冬 纪惜銮 王彦平 付咏梅 王华东 陆大祥 戚仁斌 

机构地区：[1]Department of Pathophysiology,Key Laboratory of State Administration of Traditional Chinese Medicine of the People's Republic of China,School of Medicine,Jinan University [2]Clifford Hospital,Guangzhou University of Chinese Medicine [3]High Magnetic Field Laboratory,Chinese Academy of Sciences

出　　处：《Chinese Journal of Integrative Medicine》2016年第5期353-361,共9页中国结合医学杂志（英文版）

基　　金：Supported by the Natural Science Foundation of Guangdong Province,China(No.2014A030313394);the Project of Science and Technology of Guangzhou,China(No.2014J4100098);the National Program on Key Basic Research Project(973 Program,No.2011CB707501);the Fundamental Research Funds for the Central Universities in China(No.21613401)

摘　　要：Objective： To investigate the effect and the potential mechanism of Senegenin （Sen） against injury induced by hypoxia/reoxygenation （H/R） in highly differentiated PC12 cells. Methods： The cultured PC12 cells were treated with H/R in the presence or absence of Sen （60 μmol/L）. Four groups were included in the experiment： control group, H/R group, H/R＋Sen group and Sen group. Cell viability of each group and the level of lactate dehydrogenase （LDH） in culture medium were detected for the pharmacological effect of Sen. Hoechst 33258 staining and annexin V/propidium iodide double staining were used to analyze the apoptosis rate. Moreover, mitochondrial membrane potential （△ψm）, reactive oxygen species （ROS） and intracellular free calcium （[Ca2＋]i） were measured by fluorescent staining and flow cytometry. Cleaved caspase-3 and activity of NADPH oxidase （NOX） were determined by colorimetric protease assay and enzyme linked immunosorbent assay, respectively. Results： Sen significantly elevated cell viability （P〈0.05）, decreased the leakage of LDH （P〈0.05） and apoptosis rate （P〈0.05） in H/R-injured PC12 cells. Sen maintained the value of △ψm （P〈0.05） and suppressed the activity of caspase-3 （P〈0.05）. Moreover, Sen reduced ROS accumulation （P〈0.05） and [Ca2＋]i increment （P〈0.05） by inhibiting the activity of NOX （P〈0.05）. Conclusion： Sen may exert cytoprotection against H/R injury by decreasing the levels of intracellular ROS and [Ca2＊]~, thereby suppressing the mitochondrial pathway of cellular apoptosis.Objective： To investigate the effect and the potential mechanism of Senegenin （Sen） against injury induced by hypoxia/reoxygenation （H/R） in highly differentiated PC12 cells. Methods： The cultured PC12 cells were treated with H/R in the presence or absence of Sen （60 μmol/L）. Four groups were included in the experiment： control group, H/R group, H/R＋Sen group and Sen group. Cell viability of each group and the level of lactate dehydrogenase （LDH） in culture medium were detected for the pharmacological effect of Sen. Hoechst 33258 staining and annexin V/propidium iodide double staining were used to analyze the apoptosis rate. Moreover, mitochondrial membrane potential （△ψm）, reactive oxygen species （ROS） and intracellular free calcium （[Ca2＋]i） were measured by fluorescent staining and flow cytometry. Cleaved caspase-3 and activity of NADPH oxidase （NOX） were determined by colorimetric protease assay and enzyme linked immunosorbent assay, respectively. Results： Sen significantly elevated cell viability （P〈0.05）, decreased the leakage of LDH （P〈0.05） and apoptosis rate （P〈0.05） in H/R-injured PC12 cells. Sen maintained the value of △ψm （P〈0.05） and suppressed the activity of caspase-3 （P〈0.05）. Moreover, Sen reduced ROS accumulation （P〈0.05） and [Ca2＋]i increment （P〈0.05） by inhibiting the activity of NOX （P〈0.05）. Conclusion： Sen may exert cytoprotection against H/R injury by decreasing the levels of intracellular ROS and [Ca2＊]~, thereby suppressing the mitochondrial pathway of cellular apoptosis.

关 键 词：SENEGENIN hypoxiaJreoxygenation PC12 cells reactive oxygen species NAPDH oxidase 

分 类 号：R285.5[医药卫生—中药学]