摘要
目的:探讨PI3K/AKT/mTOR信号通路对甘氨鹅脱氧胆酸钠(GCDC)诱导的人肝正常细胞HL-7702增殖、凋亡的影响。方法:将HL-7702细胞分为对照组、GCDC组(1 mmol/L的GCDC预处理细胞4 h)、GCDC+LY294002组(GCDC处理细胞4 h后,使用15μmol/L的LY294002处理细胞24 h)和GCDC+IGF-1组(GCDC处理细胞4 h后,使用20 nmol/L的IGF-1处理细胞24 h)。处理结束后,通过Western blot检测PI3K、AKT、p-AKT、mTOR、p-mTOR、Bcl-2、Bax和Nrf2蛋白表达;MTT法及Annexin V-FITC/PI双染法检测细胞增殖和凋亡率;DCFH-DA探针法检测细胞活性氧(ROS)含量。结果:GCDC可明显下调HL-7702细胞PI3K、p-AKT、p-mTOR、Bcl-2和Nrf2表达,上调Bax表达,抑制细胞增殖,诱导凋亡,诱导细胞ROS产生(P<0.05),LY294002可增强GCDC对HL-7702细胞增殖、凋亡、ROS水平及PI3K、p-AKT、p-mTOR、Bcl-2、Bax和Nrf2表达影响,IGF-1反之。结论:激活PI3K/AKT/mTOR信号通路可改善胆汁性肝纤维化,机制可能与抗凋亡和抗氧化有关。
AIM:To investigate the effects of PI3K/AKT/mTOR signaling pathway on proliferation and apoptosis of human hepatocyte HL-7702 induced by GCDC.METHODS:HL-7702 cells were divided into control group,GCDC group(1mmol/L GCDC pretreated cells for 4 hours),GCDC+LY294002 group(GCDC treated cells for 4 hours,LY294002 treated cells for 24 hours)and GCDC+IGF-1 group(GCDC treated cells for 4 hours,20nmol/L of IGF-1 treated cells for 24 hours).After treatment,the expressions of PI3K,AKT,p-AKT,mTOR,p-mTOR,Bcl-2,Bax and Nrf2 were detected by Western blot;the proliferation and apoptotic rates were detected by MTT and Annexin V-FITC/PI double staining,respectively.The content of ROS was detected by fluorescent DCFH-DA probe.RESULTS:GCDC could significantly down-regulate the expression of PI3K,p-AKT,p-mTOR,Bcl-2 and Nrf2 in HL-7702 cells,up-regulate the expression of Bax,inhibit cell proliferation,induce apoptosis and induce ROS production(P﹤0.05).LY294002 could enhance the effect of GCDC on proliferation,apoptosis,ROS level and PI3K,p-AKT,p-mTOR,Bcl-2,Bax and Nrf2 expression of HL-7702 cells;IGF-1 vice versa.CONCLUSION:Activation of PI3K/AKT/mTOR signaling pathway can improve bile liver fibrosis,and the mechanism may be related to anti-apoptosis and anti-oxidation.
作者
翁丹丹
王俊波
郦光晓
WENG Dandan;WANG Junbo;LI Guangxiao(Department of Infection,Yuyao People's Hospital,Yuyao 315400,Zhejiang,China;Zhejiang University City College School of Medicine,Hangzhou 310015,Zhejiang,China;Sir Run Run Shaw Hospital,Zhejiang University School of Medicine,Hangzhou 310016,Zhejiang,China)
出处
《中国临床药理学与治疗学》
CAS
CSCD
2019年第2期121-127,共7页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
浙江省药学会医院药学专项科研资助课题(WWQV77068)
