Effect of autophagy inhibitor chloroquine on acute alcoholinduced liver disease

doi:10.12047/j.cjap.5551.2018.025

Abstract

Abstract: Objective:To investigate the role of autophagy inhibitor chloroquine (CQ) in acute ethanol-induced liver injury and its mechenism. Methods:Twenty-one C57BL/6 male mice were randomly divided into three groups:control group, ethanol group, CQ + ethanol group (n=7). Mice in ethanol group were administered 33% (v/v) ethanol at a dose of 4.5 g/kg body weight. Ethanol-induced liver steatosis in each group was detected by hematoxylin and eosin staining. Hepatic lipid accumulation was detected by staining with Oil red O. Hepatic tissue triglyceride (TG) levels, serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) were determined by biochemical assays. Protein expression of microtubule-associated protein 1 light chain 3(LC3) and nuclear factorκB p65(NF-κB p65) were measured by Western blot and immunofluorescence. Pro-inflammatory factors tumor necrosis factor-α(TNF-α)、interleukin 6(IL-6) were detected by ELISA. Results:Compared with control group, ethanol induced liver injury proved by accumulation of hepatic lipids, TG levels, AST and ALT activities were significantly increased by ethanol, protein expression of LC3-Ⅱ was also markedly increased by ethanol. Compared with ethanol group, addition of CQ increased furtherthe level of LC3-Ⅱexpression, and TG amount, serum AST and ALT activities, and the expression of NF-κB p65, TNF-αand IL-6. Conclusion:Acute ethanol-intake could induce liver steatosis and inflammation, and autophagy inhibitor CQ exacerbatedethanol-induced liver injury, suggested that autophagy might be protective effect in acute ethanol-induced liver disease.

Key words: ethanol, autophagy, chloroquine, liver injury, mouse

SANG Wen-hua, ZENG Mei-Chun, CHEN Sha, CHEN Ran, FAN Xiao-fang, GONG Yong-sheng, ZHANG Hai-lin, ZHANG Hong-yu, KONG Xiao-xia. Effect of autophagy inhibitor chloroquine on acute alcoholinduced liver disease[J]. CJAP, 2018, 34(2): 102-105.

References

[1] Pan JH, Lim YJ, Kim JH, et al. Root bark of Ulmus davidiana var. japonica restrains acute alcohol-induced hepatic steatosis onset in mice by inhibiting ROS accumulation[J]. PLoS One, 2017, 12(11):e0188381.
[2] Tilg H, Moschen AR, Szabo G. Interleukin-1 and inflammasomes in alcoholic liver disease/acute alcoholic hepatitis and nonalcoholic fatty liver disease/nonalcoholic steatohepatitis[J]. Hepatology, 2016, 64(3):955-965.
[3] Ju C, Mandrekar P. Macrophages and alcohol-related liver inflammation[J]. Alcohol Res, 2015, 37(2):251-262.
[4] Dunn WA Jr. Autophagy and related mechanisms of lysosome-mediated protein degradation[J]. Trends Cell Biol, 1994, 4(4):139-143.
[5] Glick D, Barth S, Macleod KF. Autophagy:cellular and molecular mechanisms[J]. J Pathol, 2010, 221(1):3-12.
[6] 朱焕勉, 陈然, 薛峰, 等. 自噬抑制剂氯奎对低氧诱导肺动脉平滑肌细胞增殖的影响[J]. 中国应用生理学杂志, 2014, 30(1):8-12.
[7] Hetz C. Apoptosis, necrosis and autophagy:from mechanisms to biomedical applications[J]. Curr Mol Med, 2008, 8(2):76-77.
[8] Mariйo G, Lópezotín C. Autophagy:molecular mechanisms, physiological functions and relevance in human pathology[J]. Cell Mol Life Sci, 2004, 61(12):1439-1454.
[9] 何云凌, 吴丽颖, 黄欣, 等. 自噬对不同低氧条件下PC12细胞存活的影响[J]. 中国应用生理学杂志, 2013, 29(3):193-196.
[10] Ding WX, Manley S, Ni HM. The emerging role of autophagy in alcoholic liver disease[J]. Exp Biol Med, 2011, 236(5):546-556.
[11] Ni HM, Du K, You M, et al. Critical role of FoxO3a in alcohol-induced autophagy and hepatotoxicity[J]. Am J Pathol, 2013, 183(6):1815-1825.
[12] Ding WX, Li M, Chen X, et al. Autophagy reduces acute ethanol-induced hepatotoxicity and steatosis in mice[J]. Gastroenterology, 2010, 139(5):1740-1752.
[13] Han JJ, Plawecki MH, Doerschuk PC, et al. Ordinary diffe rential equation models for ethanol pharmacokinetic based on anatomy and physiology[C]//Engineer Med Biol Society, 2006. International Conference of the IEEE. IEEE, 2006:5033-5036.
[14] Lieber CS. Alcoholic fatty liver:its pathogenesis and mechanism of progression to inflammation and fibrosis[J]. Alcohol, 2004, 34(1):9-19.
[15] Lu Y, Cederbaum AI. CYP2E1 and oxidative liver injury by alcohol[J]. Free Radic Biol Med, 2008, 44(5):723-738.
[16] Tak PP, Firestein GS. NF-kappa B:a key role in inflammatory diseases[J]. J Clin Invest, 2001, 107(1):7-11.
[17] da silva BS, Rodrigues GB, Rocha SW, et al. Inhibition of NF-κB activation by diethylcarbamazine prevents alcohol-induced liver injury in C57BL/6 mice[J]. Tissue Cell, 2014, 46(5):363-371.
[18] Mizushima N, Yoshimori T, Levine B. Methods in mammalian autophagy research[J]. Cell, 2010, 140(3):313326.
[19] Kong X, Yang Y, Ren L, et al. Activation of autophagy attenuates EtOH-LPS-induced hepatic steatosis and injury through MD2 associated TLR4 signaling[J]. Scient Rep, 2017, 7(1):9292-9304.
[20] Cho HI, Choi JW, Lee SM. Impairment of autophagosomelysosome fusion contributes to chronic ethanol-induced liver injury[J]. Alcohol, 2014, 48(7):717-725.
[21] Chung KW, Kim KM, Choi YJ, et al. The critical role played by endotoxin-induced liver autophagy in the maintenance of lipid metabolism during sepsis[J]. Autophagy, 2017, 13(7):1113-1129.