Nodosin通过Apaf-1和caspase-3诱导HepG2细胞凋亡  

Nodosin induces apoptosis of HepG2 cells by increasing Apaf-1 expression and activating caspase-3

作　　者：海广范[1] 张慧[1] 马敬[2] 郭兰青[3] 王海燕[4] HAI Guang-fan;ZHANG Hui;MA Jing;GUO Lan-qing;WANG Hai-yan(Pharmacy School, Xinxiang Medical University, Xinxiang 453003, China;The Second Affiliated Hospital, Xinxiang Medical University, Xinxiang 453003, China;The Nursing School, Xinxiang Medical University, Xinxiang 453003, China;The First Affiliated Hospital, Xinxiang Medical University, Xinxiang 453003, China)

机构地区：[1]新乡医学院药学院,河南新乡453003 [2]新乡医学院第二附属医院,河南新乡453003 [3]新乡医学院护理学院,河南新乡453003 [4]新乡医学院第一附属医院,河南新乡453003

出　　处：《中国病理生理杂志》2018年第12期2193-2197,共5页Chinese Journal of Pathophysiology

基　　金：河南省科技攻关计划项目(No.122102310286)

摘　　要：目的:研究传统中药提取物nodosin对人肝细胞癌Hep G2细胞凋亡的影响,并探讨其作用机制。方法:将nodosin设置为1. 25μmol/L、2. 5μmol/L、5μmol/L、10μmol/L和20μmol/L不同浓度组,作用于Hep G2细胞24 h后,用Hoechst 33258染色和电镜观察不同浓度的药物对细胞形态学的影响,用流式细胞术检测细胞凋亡率,用RT-qPCR检测凋亡蛋白酶激活因子1 (Apaf-1) mRNA的表达,用Western blot检测caspase-3及其前体和活化体的蛋白水平。结果:形态学结果显示,随着用药剂量的增加,细胞皱缩和细胞核偏移越明显,凋亡小体在5μmol/L、10μmol/L和20μmol/L剂量组明显增多。Apaf-1 mRNA的表达增加,caspase-3及其前体的表达和cleaved caspase-3的蛋白水平随着用药剂量的增加逐渐增加(P <0. 01)。结论:Nodosin能够诱导Hep G2细胞凋亡。该作用可能是通过增加Apaf-1 mRNA的表达继之激活caspase-3实现的。AIM:To investigate the effects of nodosin on apoptosis of human hepatocellular carcinoma HepG2cells.METHODS:HepG2cells were treated with nodosin at different concentrations(1.25μmol/L,2.5μmol/L,5μmol/L,10μmol/L and20μmol/L)for24h.The morphological changes of the HepG2cells were observed by Hoechst33258staining and electron microscopy.The apoptotic rates were analyzed by flow cytometry.The mRNA expression of apoptotic protease-activating factor-1(Apaf-1)was detected by RT-qPCR.The protein levels of pro-caspase-3,caspase-3and cleaved caspase-3were determined by Western blot.RESULTS:HepG2cells showed obvious cell shrinkage and nucleus drift when treated with nodosin as the concentration was increased.Many apoptotic bodies were observed in5μmol/L,10μmol/L and20μmol/L nodosin groups.The mRNA expression of Apaf-1was increased in5μmol/L,10μmol/L and20μmol/L nodosin groups as compared with control group(P<0.05).The protein levels of pro-caspase-3,caspase-3and cleaved caspase-3were increased with the increasing dose of nodosin(P<0.05).CONCLUSION:Nodosin induces the apoptosis of HepG2cells.This effect was related to increasing Apaf-1mRNA expression and subsequently promoting the activation of caspase-3.

关 键 词：肝细胞癌 NODOSIN 细胞凋亡 CASPASE-3 凋亡蛋白酶激活因子1 

分 类 号：R735.7[医药卫生—肿瘤] R363.2[医药卫生—临床医学]