黄连红曲药对调控Nrf2/HO-1通路改善非酒精性脂肪肝的作用研究

Effects of Huanglian(黄连)-Hongqu(红曲)Herb Pair on Improving Nonalcoholic Fatty Liver Disease via Regulating Nrf2/HO-1 Pathway

目的:研究黄连红曲对非酒精性脂肪肝(Nonalcoholic Fatty Liver Disease, NAFLD)的疗效及机制。方法:62只SD大鼠随机分成正常对照组(12只)和造模组(50只),采用高脂饲料加脂肪乳灌胃造模8 w,造模成功后将模型组随机分为模型对照组、黄连红曲1.1、2.1 g/kg组和奥贝胆酸0.5 mg/kg组,每组8只。采用全自动生化仪检测血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白(LDL-C),血糖(GLU)含量,谷丙转氨酶(ALT)、天冬氨酸转氨酶(AST)活力;比色法检测血清丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GSH-Px)含量;生化法测定肝脏TC、TG、FFA含量;ELISA法检测血清胰岛素含量;HE和油红O染色分别观察肝脏病理和脂质沉积情况;qPCR检测肝脏Nrf2和Ho1 mRNA表达;免疫组化检测肝脏Nrf2的表达;WB检测肝脏Nrf2、HO-1蛋白表达情况。结果:与正常对照组比较,模型对照组大鼠体质量增加,肝指数增加,血脂TC、TG、LDL-C含量和肝脏脂质TC、TG、FFA含量显著升高,血清ALT、AST活力升高,血清MDA含量增加,SOD活力、GSH-Px含量降低,胰岛素抵抗指数增加,肝脏出现脂肪样变及大量脂滴,Nrf2、Ho1 mRNA及蛋白表达明显下调(P<0.05或P<0.01);与模型对照组比较,黄连红曲组大鼠体质量减轻,血脂降低,肝脏脂质含量降低,肝功能得到改善,胰岛素抵抗指数降低,肝脏病理学显示大鼠肝脏脂质沉积减轻,脂滴减少;肝脏Nrf2、Ho1 mRNA和蛋白表达明显上调(P<0.05或P<0.01)。结论:黄连红曲药对可以改善大鼠NAFLD,其机制可能与上调Nrf2/HO-1通路减轻氧化应激损伤有关。

Objective:To investigate the therapeutic effects of Huanglian(黄连)-Hongqu(红曲)herb pair on non-alcoholic fatty liver disease(NAFLD)and its mechanism.Methods:Sixty-two SD rats were randomly divided into a blank control group(12 rats)and a model control group(50 rats).High-fat diet and fat milk were used to make models by gavage for 8 weeks.After successful modeling,the model group was randomly divided into a model control group,Huanglian-Hongqu 1.1 g/kg and 2.1 g/kg groups,and an obeticholic acid 0.5 mg/kg group,with 8 rats in each group.Automatic biochemical analyzer was used to detect the content of serum total cholesterol(TC),triglyceride(TG),lowdensity lipoprotein(LDL),blood sugar(GLU),alanine aminotransferase(ALT),and aspartate aminotransferase(AST).The content of serum malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione reductase(GSH-Px)was determined by colorimetry.Biochemical method was used to determine the liver content of TC,TG,and free fatty acid(FFA).The enzyme-linked immunosorbent assay(ELISA)was used to detect serum insulin level.The hematoxylin-eosin(HE)staining and oil red O staining were adopted to observe liver pathology and lipid deposition,respectively.Real-time quantitative polymerase chain reaction(qPCR)was used to detect the mRNA expression levels of liver nuclear respiratory factor-2(Nrf2)and heme oxygenase 1(Ho-1).Immunohistochemistry was used to detect the expression of liver Nrf2,while Western blotting was used to detect the protein expression levels of liver Nrf2 and HO-1.Results:Compared with those in the blank control group,rats in the model control group gained weight and increased the liver index.The serum content of TC,TG,and LDL-C and liver lipid content of TC,TG,and FFA were significantly up-regulated in the model control group,the ALT and AST activities were increased,the MDA content was increased,the SOD,and GSH-Px activities were decreased,insulin resistance index was increased,and fat-like changes and a large number of lipid droplets were observed in liver tissues(P<0.05 or P<0.01).As compared with the model control group,the body weight,blood lipids,and liver lipid levels were decrease in Huanglian-Hongqu groups,the liver function was improved,and insulin resistance index was decreased.Meanwhile,liver pathology showed that liver lipid deposition and lipid droplets were reduced in rats in Huanglian-Hongqu groups,and the mRNA and protein expression levels of hepatic Nrf2 and Ho-1 were significantly up-regulated(P<0.01 or P<0.05).Conclusion:Huanglian-Hongqu herb pair can improve NAFLD in rats,and its mechanism may be related to the up-regulation of the Nrf2/HO-1 pathway to reduce oxidative stress injury.