中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (在线): 1-8.

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抑制NOX缓解酒精性肝细胞损伤模型小鼠肝细胞损伤及脂代谢紊乱

崔玮1，崔  迪1，欧阳婷2，李  想1，位会亭1，薛崴月1，周  刚1，邱  烨2   

  1. 1湖南大学体育学院，湖南省长沙市  410000；2湖南大学生物学院，湖南省长沙市  410000
  • 收稿日期:2021-12-21 修回日期:2022-01-29 出版日期:2022-01-08 发布日期:2022-02-09
  • 通讯作者: 崔迪，博士，助理教授，湖南大学体育学院人体科学实验室，湖南省长沙市 410000
  • 作者简介:崔玮，女，1996年生，山东省淄博市人，汉族，湖南大学在读硕士，主要从事运动对健康促进的分子机制研究
  • 基金资助:
    湖南省体育局课题(2020XH030)，项目负责人：崔玮

Inhibiting NOX alleviates alcoholic liver damage and lipid metabolism disorder

Cui Wei1, Cui Di1, Ouyang Ting2, Li Xiang1, Wei Huiting1, Xue Weiyue1, Zhou Gang1, Qiu Ye2   

  1. 1Department of Physical Education, Hunan University, Changsha 410000, Hunan Province, China; 2School of Biology, Hunan University, Changsha 410000, Hunan Province, China)
  • Received:2021-12-21 Revised:2022-01-29 Online:2022-01-08 Published:2022-02-09
  • Contact: Cui Di, PhD, Department of Physical Education, Hunan University, Changsha 410000, Hunan Province, China
  • About author:Cui Wei, Department of Physical Education, Hunan University, Changsha 410000, Hunan Province, China
  • Supported by:
    the Project of Hunan Provincial Sports Bureau, No.2020XH030 (to CW)

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摘要： 文题释义：
还原型尼克酰胺腺嘌呤二核苷酸磷酸氧化酶(NADPH oxidase，NOX)：作为一种多酶复合物在多种细胞类型中均有表达，是胞内以NADPH为底物生成活性氧的主要酶类之一。NOX激活参与调控细胞氧化应激，对机体宿主防御及炎症反应起关键作用。
酒精性肝损伤(alcoholic liver damage，ALD)：由于长期或大量饮酒导致的肝脏细胞结构异常、脂肪堆积、脂代谢紊乱和肝功能障碍性损伤。
氧化应激：指体内氧化与抗氧化作用失衡的一种状态，参与肝损伤的发生发展。许多危险因素，包括酒精、药物、环境污染和辐射，都可引起肝组织氧化应激。

摘要
背景：研究表明，应激状态下小鼠心肌、骨骼肌、肾脏及脑组织NOX蛋白过度上调，而NOX介导氧化应激在过量饮酒导致的酒精性肝损伤作用尚不清楚。
目的：探讨NOX介导氧化应激在肝脏正常生理及酒精性肝损伤病理状态下的作用。
方法：①6只雄性C57BL/6J小鼠随机分为正常对照组、正常药物组，每组3只；药物组腹腔注射NOX抑制剂夹竹桃麻素(10 mg/kg)，正常对照组注射同等剂量溶剂，连续注射4 d；检测小鼠血清及肝组织脂代谢相关指标，并检测肝组织NOX2、NOX4蛋白表达；②18只C57BL/6J雄性小鼠随机分为普通对照组、饮酒对照组、饮酒药物组，每组6只；使用Lieber-DeCarli液体饲料构建酒精性肝细胞损伤模型，普通对照组喂养TP4030C对照液体饲料，饮酒对照组和饮酒药物组喂养TP4030B酒精液体饲料，饮酒药物组在造模期间腹腔注射夹竹桃麻素(10 mg/kg，6 d/周，连续5周)，其他两组小鼠给予同等剂量溶剂；③记录小鼠体质量、附睾脂肪相对质量、肝指数、骨骼肌相对质量，进行糖耐量实验，测定血清肝功能损伤及脂代谢指标，检测肝组织丙二醛、超氧化物歧化酶水平及三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇水平，观察肝组织苏木精伊红染色情况，测试肝组织NOX2、NOX4蛋白表达。
结果与结论：①与正常对照组比，正常药物组小鼠肝组织三酰甘油水平、血清谷丙转氨酶及谷草转氨酶水平显著增加，肝组织高密度脂蛋白胆固醇水平及NOX2、NOX4蛋白表达显著降低(均P < 0.05或P < 0.01)；②与普通对照组比，饮酒对照组小鼠附睾脂肪相对质量、血清及肝组织高密度脂蛋白胆固醇含量显著降低，肝组织三酰甘油、低密度脂蛋白胆固醇、血清谷丙转氨酶水平及NOX4蛋白表达显著增加；③与饮酒对照组比，饮酒药物组小鼠血清及肝组织高密度脂蛋白胆固醇水平显著增加，血清谷丙转氨酶、肝组织低密度脂蛋白胆固醇水平和NOX4蛋白表达显著降低；④结果证实，抑制NOX缓解酒精性肝细胞损伤及脂代谢紊乱，夹竹桃麻素在肝脏生理及酒精性肝损伤病理状态下作用不同。
https://orcid.org/0000-0002-5937-0343 (崔玮)

关键词: 酒精性肝损伤, 氧化应激, NADPH氧化酶, NOX4, 脂代谢, 夹竹桃麻素

Abstract: BACKGROUND: Studies have shown NOX protein expression up-regulated in myocardium, skeletal muscle, kidney, and brain under stess conditions, while its role in alcoholic liver damage pathology was unclear.
OBJECTIVE: To investigate the role of NOX mediated oxidative stress under liver normal physiology and alcoholic liver damage pathology.
METHODS: 6 Male C57BL/6J mice were randomly divided into normal control group and normal drug group, with 3 mice in each group. Mice in normal drug group were intraperitoneally injected by 10 mg/Kg Apocynin for 4 consecutive days, while normal control group was injected with the same dose of solvent. Lipid metabolism related indexes in serum and liver tissues of mice were detected, and the protein expressions of NOX2 and NOX4 in liver tissues were determined. 18 Male C57BL/6J mice were randomly divided into common group, alcoholic liver damage (ALD) group and ALD combined with drug group, with 6 mice in each group. ALD model was constructed by Lieber-DeCarli liquid diet with TP4030C in common group as well as TP4030B in ALD group and ALD combined with drug group. Mice in ALD combined with drug group was intraperitoneally injected with 10 mg/Kg Apocynin 6 days a week for 6 weeks, while mice in other two groups were injected with the same dose of solvent. The mice body weight, epididymal fat relative content, liver index and skeletal muscle relative content, was either recorded or calculated. Glucose tolerance test was used to detect mice insulin sensitivity, and the serum liver function damage (alanine aminotransferase, ALT and Aspartate aminotransferase, AST) and lipid metabolism indexes were measured by commercial chemical kits. The liver tissue malondialdehyde (MDA), superoxide dismutase (SOD), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) was determined by colorimetry. The change of hepatocyte damage was observed by using HE staining. Western blot was applied to detect NOX2, and NOX4 protein expression in liver tissue. Ethic approval was obtained from the Experimental Animal Ethics Committee of Hunan University in China (approval No. HNUBI0202101007).
RESULTS AND CONCLUSION: Compared with normal control group, the content of TG in liver tissue and the levels of serum ALT and AST increased significantly in normal drug group; the level of HDL-C and the expressions of NOX2 and NOX4 decreased evidently in liver tissue. Compared to common group, the relative content of epididymal fat, as well as the content of HDL-C in both serum and liver tissue were strikingly decreased in ALD group and the level of serum ALT increased; the levels of TG, LDL-C and the protein expression of NOX4 were particularly increased in liver tissue. Compared to ALD group, the levels of HDL-C in both serum and liver tissue increased notably in ALD combined with drug group ; the serum ALT level, the content of LDL-C in liver and the protein expression of NOX4 decreased remarkably. In conclusion, inhibiting NOX alleviates hepatocyte damage and lipid metabolism disorder; Apocynin has different effects on liver physiology and pathological state of ALD.

Key words: alcoholic liver damage, oxidative stress, NADPH oxidase, NOX4, lipid metabolism, apocynin

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