N-乙酰半胱氨酸对非酒精性脂肪性肝炎模型小鼠肝细胞线粒体自噬的影响

肝脏 ›› 2020, Vol. 25 ›› Issue (4): 382-386.

N-乙酰半胱氨酸对非酒精性脂肪性肝炎模型小鼠肝细胞线粒体自噬的影响

张宁萍, 方颖, 刘雪静, 谢黎, 吴健, 沈锡中

200032 上海复旦大学附属中山医院消化科上海市肝病研究所(张宁萍,方颖,吴健,沈锡中);复旦大学基础医学院病原生物系(刘雪静,谢黎,吴健)

收稿日期:2020-04-05 发布日期:2020-05-09
通讯作者: 沈锡中,Email:shen.xizhong@zs-hospital.sh.cn;吴健,Email:jian.wu@fudan.edu.cn
作者简介:共同第一作者:方颖
基金资助:
北京医卫健康公益基金会(YWJKJJHKYJJ-G17014)

Effect of N-acetylcysteine on mitophagy of hepatocytes in mice with nonalcoholic steatohepatitis

ZHANG Ning-ping^1,2,*, FANG Ying^1,2, XIE Li³, WU Jian^1,2,3, SHEN Xi-zhong^1,2

1. Department of Gastroenterology and Hepatology, Zhongshan Hospital of Fudan University, Shanghai 20032, China;
2. Shanghai Institute of Liver Diseases, Fudan University Shanghai Medical College, Shanghai 20032, China;
3. Department of Medical Microbiology, Fudan University School of Basic Medical Sciences, Shanghai 200032, China

Received:2020-04-05 Published:2020-05-09
Contact: SHEN Xi-Zhong, Email: shen.xizhong@zs-hospital.sh.cn;WU Jian, Email: jian.wu@fudan.edu.cn

摘要/Abstract

摘要： 目的在非酒精性脂肪性肝炎模型小鼠中探索N-乙酰半胱氨酸(NAC)对肝细胞线粒体自噬的影响。方法 C57BL/J6小鼠分别给予16周的正常饮食、高脂高糖(HFCD)饮食和HFCD+NAC饮食。比较HE和Masson染色、ALT、AST、IL-1β、肝组织甘油三酯水平评价小鼠肝损伤。通过免疫荧光染色观察线粒体自噬。比较3组小鼠肝组织内线粒体自噬标志物在mRNA和蛋白水平的变化。结果 HFCD组小鼠较对照组ALT[(24.9±2.12)比(176.7±44.32)U/L,P<0.05]、AST[(76.7±9.06)比(291.3±39.66)U/L,P<0.05]、IL-1β[(2.94±0.08)比(9.12±1.21),P<0.05]显著升高,HFCD+NAC组较HFCD组肝功能好转,IL-1β降低[(9.12±1.21)比(6.77±0.58)ng/L,P<0.05]。HFCD组小鼠肝组织内Parkin、PINK1表达降低,同时LC3B II/I比值降低,P62表达量增高。提示HFCD组小鼠肝细胞线粒体自噬水平降低。HFCD+NAC组较HFCD组线粒体自噬水平升高,差异有统计学意义(P<0.05)。结论 NAC可能通过改善肝细胞线粒体自噬,进而减轻肝细胞炎症进展。

关键词: N-乙酰半胱氨酸, 非酒精性脂肪性肝炎, 线粒体自噬

Abstract: Objective To investigate the effect of N-acetylcysteine (NAC) on mitophagy in mice with nonalcoholic steatohepatitis.Methods C57BL/J6 mice were fed with control diet, high-fat and high-calories (HFCD) diet and a HFCD+ NAC diet for 16 weeks. HE and Masson staining were used to evaluate liver injury. Liver function, IL-1β, blood glucose and triglyceride levels in liver tissues were detected. Immunofluorescence staining of frozen section of liver tissue was used to observe mitophagy. Changes of mRNA and protein levels of mitophagy markers in liver tissues of 3 groups of mice were compared. Results Compared to control group, the ALT [(24.9±2.12) vs (176.7±44.32)U/L, P<0.05], AST [(76.7±9.06) vs (291.3±39.66)U/L, P<0.05] and IL-1β [(2.94±0.08) vs (9.12±1.21), P<0.05] levels of HFCD group were increased. The liver function of HFCD+NAC group was improved, and the IL-1β levels were decreased [(9.12±1.21) vs (6.77±0.58)ng/L, P<0.05]. The expression of Parkin, PINK1 and LC3B II/I ratio in the liver tissues of HFCD group decreased, while the P62 level was increased. These results suggested that the level of mitophagy in HFCD group was decreased. Compared with the HFCD group, mitophagy of hepatocytes in the HFCD+NAC group improved (P<0.05).Conclusion NAC may reduce hepatocyte inflammatory progression by improving mitophagy of hepatocytes in mice with nonalcoholic steatohepatitis.

Key words: N-acetylcysteine, Nonalcoholic steatohepatitis, Mitophagy

张宁萍, 方颖, 刘雪静, 谢黎, 吴健, 沈锡中. N-乙酰半胱氨酸对非酒精性脂肪性肝炎模型小鼠肝细胞线粒体自噬的影响[J]. 肝脏, 2020, 25(4): 382-386.

ZHANG Ning-ping, FANG Ying, XIE Li, WU Jian, SHEN Xi-zhong. Effect of N-acetylcysteine on mitophagy of hepatocytes in mice with nonalcoholic steatohepatitis[J]. Chinese Hepatolgy, 2020, 25(4): 382-386.

参考文献

[1] Benedict M, Zhang X. Non-alcoholic fatty liver disease: An expanded review. World J Hepatol, 2017, 9(16): 715-732.
[2] Carrico C, Meyer JG, He W, et al. The mitochondrial acylome emerges: proteomics, regulation by sirtuins, and metabolic and disease implications. Cell Metab, 2018, 27(3): 497-512.
[3] Sedlackova L, Korolchuk V I. Mitochondrial quality control as a key determinant of cell survival. Biochim Biophys Acta Mol Cell Res, 2019, 1866(4): 575-587.
[4] Tian Z, Chen Y, Yao N, et al. Role of mitophagy regulation by ROS in hepatic stellate cells during acute liver failure. Am J Physiol Gastrointest Liver Physiol, 2018, 315(3): G374-G384.
[5] Sales I, Dzierba AL, Smithburger PL, et al. Use of acetylcysteine for non-acetaminophen-induced acute liver failure. Ann Hepatol, 2013, 12(1): 6-10.
[6] Zhang NP, Liu XJ, Xie L, et al. Impaired mitophagy triggers NLRP3 inflammasome activation during the progression from nonalcoholic fatty liver to nonalcoholic steatohepatitis. Lab Invest, 2019, 99(6): 749-763.
[7] Liu XJ, Duan NN, Liu C, et al. Characterization of a murine nonalcoholic steatohepatitis model induced by high fat high calorie diet plus fructose and glucose in drinking water. Lab Invest, 2018, 98(9): 1184-1199.
[8] Wang DW, Yin YM, Yao YM. Advances in the management of acute liver failure. World J Gastroenterol, 2013, 19(41): 7069-7077.
[9] Darweesh SK, Ibrahim MF, El-Tahawy MA. Effect of N-Acetylcysteine on mortality and liver transplantation rate in non-acetaminophen-induced acute liver failure: a multicenter study. Clin Drug Investig, 2017, 37(5): 473-482.
[10] Hardy T, Oakley F, Anstee QM, et al. Nonalcoholic fatty liver disease: pathogenesis and disease spectrum. Annu Rev Pathol, 2016, 11: 451-496.
[11] Spahis S, Delvin E, Borys JM, et al. Oxidative stress as a critical factor in nonalcoholic fatty liver disease pathogenesis. Antioxid Redox Signal, 2017, 26(10): 519-541.
[12] Sanyal AJ, Campbell-Sargent C, Mirshahi F, et al. Nonalcoholic steatohepatitis: association of insulin resistance and mitochondrial abnormalities. Gastroenterology, 2001, 120(5): 1183-1192.
[13] Kojima H, Sakurai S, Uemura M, et al. Mitochondrial abnormality and oxidative stress in nonalcoholic steatohepatitis. Alcohol Clin Exp Res, 2007, 31(1 Suppl): S61-S66.
[14] Morris EM, Rector RS, Thyfault JP, et al. Mitochondria and redox signaling in steatohepatitis. Antioxid Redox Signal, 2011, 15(2): 485-504.
[15] Klionsky DJ, Andrew T. Clinical research and autophagy. Autophagy, 2014, 10(8): 1357-1358.