淫羊藿苷对脑星形胶质细胞缺氧/复氧损伤的保护作用研究  

Study on protective effect of Icarrin on hypoxia/reoxygenation injury of cerebral astrocytes

作　　者：李伟 扶国学 程绩 LI Wei;FU Guoxue;CHENG Ji(Department of Emergency,Second Affiliated Hospital of Army Military Medical University,Chongqing 400037,China;Department of Thyroid and Hernia Surgery,First Affiliated Hospital of Gannan Medical University,Ganzhou,Jiangxi 341000,China)

机构地区：[1]陆军军医大学第二附属医院急诊科,重庆400037 [2]赣南医学院第一附属医院甲状腺疝外科,江西赣州341000

出　　处：《重庆医学》2023年第13期1932-1937,共6页Chongqing medicine

基　　金：重庆市科委技术创新与应用示范项目(cstc2018jscx-msybX0050)。

摘　　要：目的探讨淫羊藿苷对星形胶质细胞缺氧/复氧(H/R)损伤的保护作用。方法利用CCK-8法测定7种不同浓度的淫羊藿苷培养液对正常培养星形胶质细胞24 h细胞活力的影响,筛选出安全浓度范围。将星形胶质细胞分为对照组、H/R组和淫羊藿苷组。H/R组和淫羊藿苷组采用无血清缺氧缺糖后恢复血清复氧复糖培养的方法构建H/R星形胶质细胞模型,淫羊藿苷组在此基础上使用安全浓度的淫羊藿苷进行干预。CCK-8法测定各组细胞活力,筛选出淫羊藿苷最佳有效浓度。检测各组细胞中乳酸脱氢酶(LDH)、超氧阴离子自由基(·O_(2)^(-))及超氧化物歧化酶(SOD)水平,H2DCFDA荧光探针检测各组细胞内活性氧(ROS)水平,JC-1荧光探针测定各组细胞线粒体膜电位水平,流式细胞术检测各组细胞凋亡情况。结果7种浓度淫羊藿苷组中,3.8、7.5、15.0μmol/L淫羊藿苷组与对照组细胞活力比较,差异无统计学意义(P>0.05)。与对照组比较,H/R组细胞活力降低(P<0.05);与H/R组比较,3.8、7.5、15.0μmol/L淫羊藿苷组细胞活力增强(P<0.05),且呈现剂量依赖的趋势,确定15.0μmol/L为淫羊藿苷最佳有效浓度。与对照组比较,H/R组LDH、·O_(2)^(-)、细胞内ROS水平明显增加,SOD活性减弱,线粒体膜电位下降,细胞凋亡数目增多(P<0.05)。与H/R组比较,淫羊藿苷组LDH、·O_(2)^(-)、细胞内ROS水平明显减少,SOD活性增强,线粒体膜电位上升,细胞凋亡数目减少(P<0.05)。结论淫羊藿苷可减轻H/R引起的星形胶质细胞损伤和细胞氧化应激反应,减少细胞凋亡。Objective To investigate the protective effect of Icariin on hypoxia/reoxygenation(H/R)injury of astrocytes.Methods The effects of 7 different concentrations of Icariin culture solution on the 24 h cell activity of normally cultured astrocytes were determined by the CCK-8 method,and the safe concentration range was screened out.The astrocytes were divided into the control group,H/R group and Icariin group.The H/R group and Icariin group adopted the method of serum reoxygenation and reglucose culture method after serum-free hypoxia and glucose deficiency to construct the H/R astrocyte model,on this basis the Icariin group used the safe use concentration of Icariin to conduct the intervention.The cell activity in each group was detected by the CCK-8 method.The optimal Icariin effective concentration was screened out.The levels of lactate dehydrogenase(LDH),superoxide(·O_(2)^(-))and superoxide dismutase(SOD)in cells of each group was detected.The level of reactive oxygen species(ROS)in each group was detected by H2DCFDA fluorescence probe,the level of mitochondrial membrane potential was measured by JC-1 fluorescence probe,and the apoptosis of cells in each group was detected by flow cytometry.Results Among the 7 concentrations of Icariin groups,there was no statistically significant difference in cell activity between 3.8,7.5,15.0μmol/L Icariin groups and control group(P>0.05).Compared with the control group,the cell viability in the H/R group was decreased(P<0.05);compared with the H/R group,the cell viability in the 3.8,7.5 and 15.0μmol/L Icariin groups was increased(P<0.05),moreover showing the dose-dependent trend.Therefore,15.0μmol/L was determined as the best effective concentration.Compared with the control group,LDH,·O_(2)^(-)and intracellular ROS levels in the H/R group were increased significantly,the SOD activity was decreased,the mitochondrial membrane potential was decreased and the number of cell apoptosis was increased(P<0.05).Compared with the H/R group,the levels of LDH,·O_(2)^(-)and intracel

关 键 词：淫羊藿苷 星形胶质细胞 缺氧/复氧 氧化应激 凋亡 

分 类 号：R285[医药卫生—中药学]