人参皂苷通过Aldolase/AMPK/PINK1信号通路减轻油酸诱导的HL-7702细胞损伤

doi:10.12092/j.issn.1009-2501.2020.06.005

中国临床药理学与治疗学 ›› 2020, Vol. 25 ›› Issue (6): 633-639.doi: 10.12092/j.issn.1009-2501.2020.06.005

人参皂苷通过Aldolase/AMPK/PINK1信号通路减轻油酸诱导的HL-7702细胞损伤

胡文艳, 李梅, 刘华宝, 饶春燕, 李泓町

重庆市中医院肝病科,重庆 400021

收稿日期:2020-04-01 出版日期:2020-06-26 发布日期:2020-07-09
通讯作者: 李梅,女,硕士,主治中医师,研究方向:非酒精性脂肪性肝病治疗研究。Tel: 13594013750 E-mail: limei870402@126.com
作者简介:胡文艳,女,硕士,主治医师,研究方向:肝纤维化、肝硬化及肝衰竭的研究。
基金资助:
绩效激励引导专项(研发投入)(cstc2018jxj1130046)

Ginsenoside Rg1 protects HL-7702 cells against oleic acid-induced injury via aldolase/AMPK/PINK1 signalling

HU Wenyan, LI Mei, LIU Huabao, RAO Chunyan, LI Hongting

Department of Hepatology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing 400021, China

Received:2020-04-01 Online:2020-06-26 Published:2020-07-09

摘要/Abstract

摘要： 目的: 研究人参皂苷对油酸(OA)诱导人肝HL-7702细胞损伤的保护作用,及其对Aldolase/AMPK/PINK1信号通路的影响。方法: 体外培养HL-7702细胞,并将其分为对照组(C)、油酸刺激组(OA)、OA+人参皂苷组(OA+Rg1)、OA+Compound C组(OA+CC)、OA+Compound C+人参皂苷组(OA+CC+Rg1)。CCK8法和Hoechst染色法检测HL-7702细胞增殖活性,流式细胞术检测HL-7702细胞凋亡率和线粒体膜电位,ATP测定分析HL-7702细胞中ATP含量,Western blot分析HL-7702细胞中Cleaved caspase-3、PINK1、MFN2、Aldolase和p-AMPK蛋白表达水平。结果: 与C组比较,OA组细胞增殖活力明显降低(P<0.05),细胞凋亡率和促凋亡蛋白Cleaved caspase-3表达水平明显升高(P<0.05),ATP含量、线粒体膜电位(TMRE)和线粒体自噬蛋白PINK1表达水平明显降低(P<0.05);与OA组比较,OA+Rg1组细胞增殖活力明显升高(P<0.05),细胞凋亡率和Cleaved caspase-3蛋白表达水平明显降低(P<0.05),ATP含量、TMRE值、Aldolase、PINK1和p-AMPK蛋白表达水平明显升高(P<0.05);与OA+Rg1组比较,OA+CC+Rg1组HL-7702细胞增殖活力及AMPK磷酸化水平明显降低(P<0.05);Aldolase基因沉默后,Rg1不能增高细胞中PINK1和p-AMPK蛋白表达及细胞的增殖活性。结论: 人参皂苷可通过调节Aldolase/AMPK/PINK1信号通路活性改善油酸诱导的人肝HL-7702细胞损伤。

关键词: 人参皂苷, 油酸, 线粒体自噬, 肝细胞

Abstract: AIM: To study the protective effects of ginsenoside Rg1 on HL-7702 cells injury induce by oleic acid (OA), and investigate its role in aldolase/AMPK/PINK1 signalling. METHODS: HL-7702 cells were cultured in vitro and divided into five groups: control group (C), oleic acid group (OA), OA+ginsenoside (Rg1) group, OA+compound C (CC) group, and OA+CC+Rg1 group. The viability of HL-7702 cells was determined by CCK8 assay and Hoechst staining. The apoptosis and mitochondrial membrane potentials of HL-7702 cells were measured using flow cytometry. ATP content in HL-7702 cells was observed. Western blot was used to detect the expression levels of Cleaved caspase-3, PINK1, MFN2, Aldolase and p-AMPK in HL-7702 cells. RESULTS: Compared with C group, the viability of cells in OA group was significantly decreased (P<0.05), the apoptotic rate and Cleaved caspase-3 expression were greatly increased (P<0.05), ATP level, mitochondrial membrane potentials (TMRE) and PINK1 expression were significantly decreased (P<0.05). Compared with OA group, the viability of cells in OA+Rg1 group was significantly increased (P<0.05), the apoptotic rate and Cleaved caspase-3 expression were greatly decreased (P<0.05), ATP level, mitochondrial membrane potentials (TMRE) and PINK1 expression were significantly increased (P<0.05). Compared with OA+Rg1 group, the viability of cells and p-AMPK expression level in OA+CC+Rg1 group was significantly decreased (P<0.05). Reducing the expression of aldolase in cells inhibited Rg1?s actions on PINK1 and p-AMPK and cell viability. CONCLUSION: Ginsenoside Rg1 can improve the injury of HL-7702 cells via regulating aldolase/AMPK/PINK1 signaling pathway.

Key words: ginsenoside Rg1, oleic acid, mitophagy, hepatocyte

中图分类号:

R965.2

胡文艳, 李梅, 刘华宝, 饶春燕, 李泓町. 人参皂苷通过Aldolase/AMPK/PINK1信号通路减轻油酸诱导的HL-7702细胞损伤[J]. 中国临床药理学与治疗学, 2020, 25(6): 633-639.

HU Wenyan, LI Mei, LIU Huabao, RAO Chunyan, LI Hongting. Ginsenoside Rg1 protects HL-7702 cells against oleic acid-induced injury via aldolase/AMPK/PINK1 signalling[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(6): 633-639.

参考文献 19

[1]	董丽红, 张瑞芬, 黄菲, 等. 油酸诱导单纯性肝脂肪变性细胞模型的建立及应用 [J]. 中国药理学通报, 2017, 33(11): 1622-1626.
[2]	Simöes ICM, Fontes A, Pinton P, et al. Mitochondria in non-alcoholic fatty liver disease [J]. Int J Biochem Cell Biol, 2018, 95(7): 93-99.
[3]	Zhou H, Du W, Li Y, et al. Effects of melatonin on fatty liver disease: The role of NR4A1/DNA-PKcs/p53 pathway, mitochondrial fission, and mitophagy [J]. J Pineal Res, 2018, 64(1): 106-117.
[4]	毛云, 张兆瑞. 人参皂苷诱导骨髓间充质干细胞对急性肝功能衰竭大鼠肝组织再生及Wnt/β-catenin信号表达的影响 [J]. 中国临床药理学与治疗学, 2019, 22(5): 503-510.
[5]	Mao N, Tan RZ, Wang SQ, et al. Ginsenoside Rg1 inhibits angiotensin II-induced podocyte autophagy via AMPK/mTOR/PI3K pathway [J]. Cell Biol Int, 2016, 40(8): 917-925.
[6]	Gao Y, Chu S, Zhang Z, et al. Hepataprotective effects of ginsenoside Rg1- A review [J]. J Ethnopharmacol, 2017, 206(7): 178-183.
[7]	Samartsev VN, Polishchuk LS, Paydyganov AP, et al. Features of the uncoupling effect of fatty acids in liver mitochondria of mammals with different body weight [J]. Biochemistry (Mosc), 2004, 69(6): 678-686.
[8]	Roberts RF, Tang MY, Fon EA, et al. Defending the mitochondria: The pathways of mitophagy and mitochondrial-derived vesicles [J]. Int J Biochem Cell Biol, 2016, 79(4): 427-436.
[9]	Williams JA, Ding WX. Targeting pink1-parkin-mediated mitophagy for treating liver injury [J]. Pharmacol Res, 2015, 102(6): 264-269.
[10]	Hassan K, Bhalla VE, Regal ME, et al. Nonalcoholic fatty liver disease: a comprehensive review of a growing epidemic [J]. World J Gastroenterol, 2014, 20(34): 12082-12101.
[11]	Lanaka S, Hikita H, Tatsumi T, et al. Rubicon inhibits autophagy and accelerates hepatocyte apoptosis and lipid accumulation in nonalcoholic fatty liver disease in mice [J]. Hepatology, 2016, 64(6): 1994-2014.
[12]	Agalakova NI, Brailovskaya IV, Konovalova SA, et al. ATP-consuming processes in hepatocytes of river lamprey Lampetra fluviatilis on the course of prespawning starvation [J]. Comp Biochem Physiol A Mol Integr Physiol, 2016, 201(6): 95-100.
[13]	Zhang E, Yin S, Song X, et al. Glycycoumarin inhibits hepatocyte lipoapoptosis through activation of autophagy and inhibition of ER stress/GSK-3-mediated mitochondrial pathway [J]. Sci Rep, 2016, 6(4): 381-389.
[14]	Weng Z, Luo Y, Yang X, et al. Regorafenib impairs mitochondrial functions,activates AMP-activated protein kinase,induces autophagy,and causes rat hepatocyte necrosis [J]. Toxicology, 2015, 327(8): 10-21.
[15]	Zhang CS, Hawley SA, Zong Y, et al. Fructose-1,6-bisphosphate and aldolase mediate glucose sensing by AMPK [J]. Nature, 2017, 548(5): 112-116.
[16]	Lin SC, Hardie DG. AMPK: Sensing glucose as well as Cellular Energy Status [J]. Cell Metab, 2018, 27(2): 299-313.
[17]	Li Y, Zhang D, Kong L, et al. Aldolase promotes the development of cardiac hypertrophy by targeting AMPK signaling [J]. Exp Cell Res, 2018, 370(1): 78-86.
[18]	Li M, Zhang CS, Zong Y, et al. Transient Receptor Potential V Channels Are Essential for Glucose Sensing by Aldolase and AMPK[J]. Cell Metab, 2019, 30(3): 508-524.
[19]	Tian QQ, Yang Q, Wang R. Regulation mechanism of HIFs, PPARs and AMPK in hypoxic training-induced reduction of body weight [J]. Sheng Li Xue Bao, 2018, 70(5): 511-520.

人参皂苷通过Aldolase/AMPK/PINK1信号通路减轻油酸诱导的HL-7702细胞损伤

Ginsenoside Rg1 protects HL-7702 cells against oleic acid-induced injury via aldolase/AMPK/PINK1 signalling

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献 19

相关文章 15

编辑推荐

Metrics

本文评价

[1]	王坤, 许佩佩, 周兰兰, 鲁晟. 人参皂苷Rg1调控Epac1/Rap1信号通路对缺血性脑卒中大鼠神经保护作用机制研究[J]. 中国临床药理学与治疗学, 2023, 28(7): 721-727.
[2]	吴先闯, 刘瑜新, 牛玉季, 赫锦锦, 乔辉, 张蕾蕾. 肝细胞癌中DCLK1转录调控的机制研究[J]. 中国临床药理学与治疗学, 2023, 28(11): 1241-1246.
[3]	魏凤, 何阳, 樊志强, 刘世坤, 欧阳林旗. 氟非尼酮对二乙基亚硝胺诱导的肝损伤大鼠肝细胞中TGF-β1/Smads通路的抑制作用[J]. 中国临床药理学与治疗学, 2022, 27(7): 739-746.
[4]	全海燕, 江兴, 何璐. 线粒体自噬在肝胰岛素抵抗中作用机制的研究进展[J]. 中国临床药理学与治疗学, 2022, 27(2): 198-204.
[5]	叶凯丽, 郑雯, 叶啟发, 杨岚. CDK1参与肝细胞癌的发展机制及其抑制剂应用价值[J]. 中国临床药理学与治疗学, 2021, 26(9): 1086-1094.
[6]	余赛红, 郑晓亮, 蒲依依, 颜冬梅, 王孝举, 于洁. 通过核糖核苷酸还原酶M2逆转肝癌细胞对5-氟尿嘧啶耐药[J]. 中国临床药理学与治疗学, 2021, 26(7): 729-737.
[7]	宋如铮, 彭英, 王广基, 孙建国. 定量蛋白质组学常用研究技术及其在肝源性疾病发病机制中的应用进展[J]. 中国临床药理学与治疗学, 2021, 26(5): 570-578.
[8]	傅爱, 徐甜甜, 杨贞, 韩强, 窦晓兵, 李松涛. 阿魏酸抑制p38 MAPK保护棕榈酸诱导的肝细胞脂毒性[J]. 中国临床药理学与治疗学, 2021, 26(2): 137-143.
[9]	黄康，李玲，叶啟发，彭贵主. 肝细胞肝癌中长链非编码RNA LINC00844的表达及对细胞增殖和迁移能力的抑制作用[J]. 中国临床药理学与治疗学, 2020, 25(4): 366-372.
[10]	张高娇，王芯芯，周佳，王雷雷，戴勤学. 小窝蛋白Cav-1介导人参皂苷Rb1对小鼠脑缺血再灌注损伤的脑保护作用[J]. 中国临床药理学与治疗学, 2020, 25(3): 265-270.
[11]	宋姗姗, 杨洋, 何静, 唐俊峰. 人参皂苷Rg1对子痫前期大鼠氧化应激和炎症的调控作用及机制研究[J]. 中国临床药理学与治疗学, 2020, 25(10): 1088-1096.
[12]	常青青，彭英，王广基，孙建国. 抗肝癌小分子酪氨酸激酶抑制剂的临床研究进展[J]. 中国临床药理学与治疗学, 2019, 24(8): 948-956.
[13]	杨书才，张莉，刘利平，邓唯杰，周杰，刘慧，张鲍虎，金涛. 缺氧诱导因子2α对肝细胞癌化疗耐药性的影响[J]. 中国临床药理学与治疗学, 2019, 24(6): 630-636.
[14]	毛云，张兆瑞. 人参皂苷诱导骨髓间充质干细胞对急性肝功能衰竭大鼠肝组织再生及Wnt/β-catenin信号表达的影响[J]. 中国临床药理学与治疗学, 2019, 24(5): 503-510.
[15]	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.