摘要
为探讨ROS/JNK通路在槲皮素(Que)抑制异烟肼(INH)诱导L-02细胞凋亡中的作用,将L-02细胞随机分为对照组(control)、异烟肼组(INH)、槲皮素低剂量组(Que-L)和槲皮素高剂量组(Que-H).细胞培养24 h后,采用MTT法检测L-02细胞存活率;比色法测定细胞上清液中乳酸脱氢酶(LDH)活性;Hoechst 33258荧光染色法评价细胞凋亡情况;比色法检测细胞裂解液中Caspase 3活性;荧光探针DCFH-DA检测细胞线粒体ROS水平;Western blot法检测细胞中p-JNK蛋白表达水平.结果发现:与对照组比较,INH组的细胞存活率明显降低(P<0.01),而LDH活性、细胞凋亡率、Caspase 3活性明显升高(P<0.01),细胞线粒体ROS水平及细胞中p-JNK蛋白表达明显增高(P<0.01);与INH组比较,经槲皮素处理后细胞存活率明显增高,而LDH活性、细胞凋亡率及Caspase 3活性明显降低(P<0.05或P<0.01),细胞线粒体ROS水平及细胞中p-JNK蛋白表达明显降低(P<0.05或P<0.01),槲皮素高剂量组的效应更明显.可见,槲皮素对INH诱导的肝细胞凋亡具有保护作用,其机制可能与其抑制ROS/JNK通路有关.
To investigate the role of ROS/JNK pathway in quercetin(Que)inhibiting isoniazid(INH)-induced apoptosis in L-02 cells.L-02 cells were randomly divided into control group(control),isoniazid group(INH),quercetin low-dose group(Que-L)and quercetin high-dose group(Que-H).After the cells were cultured for 24 hours,the survival rate of L-02 cells was measured by MTT assay,and the activity of lactate dehydrogenase(LDH)in cell supernatant was determined by colorimetry.The apoptosis of cells was evaluated by Hoechst 33258 fluorescence staining,and the activity of Caspase 3 in cell hydrolysate was detected by colorimetry.The mitochondrial ROS level was detected by fluorescent probe DCFH-DA,and the expression of p-JNK protein in cells was measured by Western blotting.The results showed that compared with the control group,the survival rate of cells in the INH group was significantly reduced(P<0.01),while LDH activity,apoptosis rate and Caspase 3 activity were remarkable elevated(P<0.01);the mitochondrial ROS level and the expression of p-JNK protein in cells were markedly increased(P<0.01).Compared with the INH group,after quercetin treatment,the cell survival rate was significantly increased,while LDH activity,apoptosis rate and Caspase 3 activity decreased significantly(P<0.05 or P<0.01);ROS level and p-JNK protein expression reduced markedly(P<0.05 or P<0.01),and the effects of quercetin high-dose group were more obvious.In conclusion,quercetin has protective effects on INH-induced apoptosis of hepatocytes,which may be related to its inhibition of ROS/JNK pathway.
作者
陈廷玉
陈德顺
何帅兵
易燕锋
沈洪
林钜斌
卢春凤
CHEN Tingyu;CHEN Deshun;HE Shuaibing;YI Yanfeng;SHEN Hong;LIN Jubin;LU Chunfeng(School of Medicine,Huzhou University,Huzhou 313000,China;School of Internet of Things Engineering,Jiangnan University,Wuxi 214122,China)
出处
《湖州师范学院学报》
2021年第8期59-64,共6页
Journal of Huzhou University
基金
国家自然科学基金项目(81373497)
浙江省教育厅科研项目(Y201941673)。
