PI3K/AKT/mTOR信号通路对甘氨鹅脱氧胆酸钠诱导的肝细胞凋亡的机制研究

doi:10.12092/j.issn.1009-2501.2019.02.001

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (2): 121-127.doi: 10.12092/j.issn.1009-2501.2019.02.001

• 基础研究 • 下一篇

PI3K/AKT/mTOR信号通路对甘氨鹅脱氧胆酸钠诱导的肝细胞凋亡的机制研究

翁丹丹¹，王俊波²，郦光晓³

¹余姚市人民医院感染科，余姚 315400，浙江；²浙江大学城市学院医学院，杭州 310015，浙江； ³浙江大学邵逸夫医院，杭州 310016，浙江

收稿日期:2018-11-06 修回日期:2018-12-17 出版日期:2019-02-26 发布日期:2019-03-04
作者简介:翁丹丹，女，本科，住院医师，研究方向：肝病、结核病、艾滋病、发热。 E-mail：zunyu5263@163.com
基金资助:
浙江省药学会医院药学专项科研资助课题（WWQV77068）

Mechanism of PI3K/AKT/mTOR signaling pathway on apoptosis of hepatocyte induced by GCDC

WENG Dandan¹, WANG Junbo², LI Guangxiao³

¹ Department of Infection, Yuyao People's Hospital,Yuyao 315400, Zhejiang, China; ²Zhejiang University City College School of Medicine, Hangzhou 310015, Zhejiang,China; ³Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310016, Zhejiang, China

Received:2018-11-06 Revised:2018-12-17 Online:2019-02-26 Published:2019-03-04

摘要/Abstract

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15 μmol/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。

关键词: PI3K/AKT/mTOR信号通路, 甘氨鹅脱氧胆酸钠, 肝细胞损伤, 氧化应激

Abstract:

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 (1 mmol/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, 20 nmol/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.

Key words: PI3K/AKT/mTOR signaling pathway, glycochenodeoxycholate, hepatocyte injury, oxidative stress

中图分类号:

R965.2

翁丹丹，王俊波，郦光晓. PI3K/AKT/mTOR信号通路对甘氨鹅脱氧胆酸钠诱导的肝细胞凋亡的机制研究[J]. 中国临床药理学与治疗学, 2019, 24(2): 121-127.

WENG Dandan, WANG Junbo, LI Guangxiao. Mechanism of PI3K/AKT/mTOR signaling pathway on apoptosis of hepatocyte induced by GCDC[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(2): 121-127.

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PI3K/AKT/mTOR信号通路对甘氨鹅脱氧胆酸钠诱导的肝细胞凋亡的机制研究

Mechanism of PI3K/AKT/mTOR signaling pathway on apoptosis of hepatocyte induced by GCDC

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