淫羊藿苷对TRAIL诱导胶质母细胞瘤细胞凋亡的影响及其分子机制  

Effect and molecular mechanism of icariin on apoptosis of glioblastomacells induced by TRAIL

作　　者：施萌婧 臧运华 刘永吉 姜传武 刘柱 SHI Mengjing;ZANG Yunhua;LIU Yongji;JIANG Chuanwu;LIU Zhu(Shandong University of Traditional Chinese Medicine,Jinan 250014,China;不详)

机构地区：[1]山东中医药大学,济南250014 [2]青岛市海慈医疗集团

出　　处：《山东医药》2020年第29期28-31,共4页Shandong Medical Journal

基　　金：山东省青岛市产业培育计划科技惠民专项(19-6-1-13-nsh)。

摘　　要：目的观察淫羊藿苷(ICA)对肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导胶质母细胞瘤(GBM)细胞凋亡的影响,并探讨其分子机制。方法取GBM细胞U87(中度耐药)和U373(重度耐药),以不同浓度ICA、TRAIL处理,MTT法检测细胞活力确定ICA、TRAIL作用浓度分别为10μmol/L、50 ng/mL。将两种细胞各自分成对照组、TRAIL组、ICA组、TRAIL+ICA组,分别加DMSO、TRAIL、ICA、TRAIL+ICA处理48 h,采用流式细胞术测算细胞凋亡率,Western blotting法检测细胞凋亡关键蛋白Caspase-3及其切割底物PARP,ELISA法测定细胞裂解物中Caspase-3活性。将两种细胞随机分为4组,阻断组和阻断处理组先予以Caspase抑制剂z-VAD-fmk预处理4 h后再分别加入DMSO、TRAIL+ICA,对照组和处理组不预处理分别加DMSO、TRAIL+ICA,处理48 h后采用流式细胞术测算细胞凋亡率。结果与对照组、ICA组、TRAIL组比较,TRAIL+ICA组细胞凋亡率增高,细胞中Caspase-3、PARP蛋白表达量增加,细胞裂解物中Caspase-3活性增强(P均<0.05);对照组、ICA组、TRAIL组两两比较,差异均无统计学意义。处理组细胞凋亡率高于对照组、阻断组、阻断处理组(P均<0.05),对照组、阻断组、阻断处理组两两比较差异均无统计学意义。结论ICA通过Caspase依赖性方式促进TRAIL诱导的GBM细胞凋亡。Objective To observe the effect of icariin(ICA)on the apoptosis of glioblastoma(GBM)cells induced by tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)and to explore its molecular mechanism.Methods GBM cell lines U87(moderate drug resistance)and U373(severe drug resistance)were treated with different concentrations of ICA and TRAIL.The cell viability was detected by MTT.The concentrations of ICA and TRAIL were 10μmol/L and 50 ng/mL,respectively.The two kinds of cells were divided into the control group,TRAIL group,ICA group,and TRAIL+ICA group,respectively.The apoptosis rate was measured by flow cytometry.Caspase-3 and its cleavage substrate PARP were detected by Western blotting.The activity of Caspase-3 was further determined by ELISA.Two kinds of cells were randomly divided into four groups.The blocking group and the blocking treatment group were pretreated with Caspase inhibitor z-VAD-fmk(25μmol/L)for 4 h,then DMSO and TRAIL+ICA were added,respectively.After 48 hours of treatment,the apoptosis rate was measured by flow cytometry.Results Compared with the control group,ICA group or TRAIL group,the apoptosis rate,Caspase-3 and PARP protein expression and Caspase-3 activity in the cell lysate increased in the TRAIL+ICA group(all P<0.05).There was no significant difference between the control group,ICA group or TRAIL group.The apoptosis rate of the treatment group was higher than those of the control group,the blocking group and the blocking treatment group(all P<0.05);no significant difference was found between every two groups of the control group,the blocking group and the blocking treatment group.Conclusion ICA promotes TRAIL-induced apoptosis of GBM cells in a Caspase-dependent manner.

关 键 词：胶质母细胞瘤 淫羊藿苷 肿瘤坏死因子相关凋亡诱导配体 CASPASE-3活性 细胞凋亡 

分 类 号：R743[医药卫生—神经病学与精神病学]