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中的作用尚不清楚。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:(1)6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;正常药物组腹腔注射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;(2)18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;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Lieber-De Carli液体饲料构建酒精性肝细胞损伤模型,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、骨骼肌相对含量,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浓度,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浓度及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浓度显著降低,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浓度显著增加,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。
BACKGROUND:Studies have shown NADPH oxidase(NOX)protein expression will up-regulate in the myocardium,skeletal muscle,kidney,and brain under stress conditions.However,its role in alcoholic liver damage is still unclear.OBJECTIVE:To investigate the role of NOX-mediated oxidative stress in normal liver tissue and alcoholic liver damage.METHODS:(1)Six male C57 BL/6 J mice were randomly divided into normal control group and normal drug group,with three mice in each group.Mice in the normal drug group were intraperitoneally injected with 10 mg/kg apocynin for 4 consecutive days,while those in the normal control group were injected with the same dose of solvent.Lipid metabolism-related indexes in serum and liver tissue of mice were detected,and the protein expressions of NOX2 and NOX4 in liver tissue were determined.(2)Another 18 male C57 BL/6 J mice were randomly divided into common control group,alcoholic control group,alcoholic drug group,with 6 mice in each group.Mice in the common control group were given TP4030 C,while those in the other two groups were given Lieber-DeCarli liquid diet to construct animal models of alcohol-induced hepatocellular injury.During the modeling,mice in the alcoholic drug group were intrape ritoneally injected with 10 mg/kg apocynin,6 days a week,for 5 continuous weeks,while those in the other two groups were injected with the same dose of solvent.(3)Body mass,epididymal fat relative mass,liver index and relative mass of the skeletal muscle were reco rded or calculated.Glucose tole rance test was used to detect insulin sensitivity,and serum liver function and lipid metabolism indexes were measured.Levels of malondialdehyde,supe roxide dismutase,triglyce rides,highdensity lipoprotein choleste rol,and low-density lipoprotein choleste rol were determined.Pathological changes of the liver were observed using hematoxylineosin staining.Western blot assay was applied to detect NOX2 and NOX4 protein expression in liver tissue.RESULTS AND CONCLUSION:Compared with the normal control group,the content of triglycerides in liver tissue and the levels of serum alanine aminotransferase and aspartate aminotransferase in serum were significantly increased,while high-density lipoprotein choleste rol level and the expression of NOX2 and NOX4 in liver tissue decreased significantly in the normal drug group(P<0.05 or P<0.01).Compared with the common control group,the relative content of epididymal fat and the level of high-density lipoprotein cholesterol in both serum and liver tissue were significantly decreased in the alcoholic control group,while triglyceride and low-density lipoprotein choleste rol levels in liver tissue,serum alanine aminotransferase level and NOX4 protein expression were significantly increased in the alcoholic control group.Compared with the alcoholic control group,the alcoholic drug group had a significant increase in the high-density lipoprotein cholesterol levels in the serum and liver tissue but had a significant decrease in serum alanine aminotransferase level and low-density lipoprotein cholesterol level and NOX4 protein expression in liver tissue.In conclusion,inhibiting NOX alleviates alcohol-induced hepatocellular injury and lipid metabolism disorder;apocynin has different effects on liver physiological and pathological states of alcoholic liver damage.
作者
崔玮
崔迪
欧阳婷
李想
位会亭
薛崴月
周刚
邱烨
Cui Wei;Cui Di;Ouyang Ting;Li Xiang;Wei Huiting;Xue Weiyue;Zhou Gang;Qiu Ye(Department of Physical Education,Changsha 410000,Hunan Province,China;College of Biology,Hunan University,Changsha 410000,Hunan Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2022年第35期5589-5595,共7页
Chinese Journal of Tissue Engineering Research
基金
湖南省体育局课题(2020XH030),项目负责人:崔玮。
