自噬抑制剂氯喹对急性酒精诱导小鼠肝损伤的影响

doi:10.12047/j.cjap.5551.2018.025

中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (2): 102-105.doi: 10.12047/j.cjap.5551.2018.025

自噬抑制剂氯喹对急性酒精诱导小鼠肝损伤的影响

桑文华¹, 曾美纯¹, 陈莎¹, 陈然¹, 范小芳¹, 龚永生¹, 张海邻², 张宏宇³, 孔晓霞¹

1. 温州医科大学基础医学院低氧医学研究所, 浙江温州 325035;
2. 温州医科大学第二附属医院, 浙江温州 325035;
3. 温州医科大学药学院, 浙江温州 325035

收稿日期:2017-01-07 修回日期:2017-12-06 出版日期:2018-03-28 发布日期:2018-05-22
通讯作者: 孔晓霞 E-mail:kongxx@wmu.edu.cn
基金资助:
浙江省自然科学基金资助项目（LY17H010005，LY17H010009，Y17H160193）

Effect of autophagy inhibitor chloroquine on acute alcoholinduced liver disease

SANG Wen-hua¹, ZENG Mei-Chun¹, CHEN Sha¹, CHEN Ran¹, FAN Xiao-fang¹, GONG Yong-sheng¹, ZHANG Hai-lin², ZHANG Hong-yu³, KONG Xiao-xia¹

1. School of Basic Medicine, Institute of Hypoxia Research, Wenzhou 325035, China;
2. The Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou 325035, China;
3. School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou 325035, China

Received:2017-01-07 Revised:2017-12-06 Online:2018-03-28 Published:2018-05-22
Supported by:
浙江省自然科学基金资助项目（LY17H010005，LY17H010009，Y17H160193）

摘要/Abstract

摘要： 目的：探讨自噬抑制剂氯喹（CQ）对急性酒精诱导肝损伤的影响及其作用机制。方法：将雄性C57BL/6小鼠随机分为3组：正常对照组、酒精组、氯喹干预组（n=7），其中酒精组按4.5 g/kg剂量给予33%（V/V）酒精灌胃。HE和油红O染色检测各组小鼠肝组织脂滴变化；检测肝组织甘油三酯（TG）含量变化；检测血清谷草转氨酶（AST）和谷丙转氨酶（ALT）活性；免疫荧光法检测微管相关蛋白轻链3（LC3）蛋白变化；Western blot法检测LC3蛋白和核蛋白P65表达的变化；ELISA法检测促炎因子TNF-α、IL-6的变化。结果：与对照组比较，酒精组脂滴形成、TG含量、血清AST和ALT活性明显增高。与对照组比较，酒精组LC3-Ⅱ蛋白表达明显增加；与酒精组比较，氯喹干预组使酒精诱导的LC3-Ⅱ蛋白表达增强进一步加剧，使酒精诱导的TG含量、血清AST和ALT活性进一步增高，同时增加了酒精诱导的p65入核及TNFα、IL-6释放。结论：急性酒精能引起小鼠肝脏脂肪变化及炎症，而自噬抑制剂氯喹抑制自噬进程，加剧酒精诱导的肝损伤，说明自噬在酒精诱导肝损伤中可能具有保护效应。

关键词: 酒精, 自噬, 氯喹, 肝损伤, 小鼠

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

桑文华, 曾美纯, 陈莎, 陈然, 范小芳, 龚永生, 张海邻, 张宏宇, 孔晓霞. 自噬抑制剂氯喹对急性酒精诱导小鼠肝损伤的影响[J]. 中国应用生理学杂志, 2018, 34(2): 102-105.

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.

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自噬抑制剂氯喹对急性酒精诱导小鼠肝损伤的影响

Effect of autophagy inhibitor chloroquine on acute alcoholinduced liver disease

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