慢病毒介导miR-203过表达对酒精性肝病大鼠肝损伤的保护作用及其机制研究

Protection of liver injuries by lentivirus-transfected miR-203 overexpression in rats with alcohol-induced liver disease

目的探讨miR-203对酒精性肝病(ALD)大鼠肝损伤的保护作用及其作用机制。方法将60只SD大鼠随机分为ALD组(A组)、NC-miRNA/ALD组(B组)和miR-203/ALD组(C组)。将慢病毒质粒经尾静脉注射感染大鼠,并采用酒精饮料喂养法建立ALD模型,采用Realtime PCR法检测大鼠肝组织miR-203水平,常规行病理学检查,评估大鼠肝组织损伤情况。使用市售试剂盒检测大鼠肝组织匀浆超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和丙二醛(MDA)水平,采用Western Blot法检测肝组织IL-1β和NF-κB蛋白表达。结果在建模8 w末,C组大鼠肝组织miR-203水平为(2.8±0.1),显著高于B组[(1.3±0.5),P<0.05];C组动物血清AST、ALT和TBIL水平分别为(89.5±7.9)U/L、(38.5±10.1)U/L和(27.8±5.1)μmol/L,显著低于B组[(162.8±16.5)U/L、(69.6±7.5)U/L和(54.9±7.8)μmol/L,P<0.05];C组肝组织匀浆SOD和CAT水平分别为(57.6±11.4)U/mg和(54.6±9.9)U/mg,显著高于B组[(41.6±7.6)U/mg和(45.7±6.0)U/mg,P<0.05],而MDA水平为(2.8±0.1)nmol/g,显著低于B组[(5.0±0.2)nmol/g,P<0.05];Western Blot检测结果表明,C组肝组织IL-1β蛋白和NF-κB表达分别为(0.7±0.2)和(0.3±0.1),显著低于B组[(1.2±0.3)和(1.0±0.2),P<0.05]。结论miR-203过表达对ALD大鼠肝损伤起到了保护作用,其可能的机制是增强了抗氧化和抗炎作用。

Objective To explore the protective effect of liver injuries by lentivirus-transfected miR-203 overexpression in rats with alcohol-induced liver disease(ALD).Methods 60 male SD rats were randomly divided into ALD(group A),NC-miRNA plus ALD(group B)and miR-203 plus ALD(group C).Lenti-NC or Lenti-miR-203 were administered to rats by tail vein injection using a 30 gauge ultra-fine insulin syringe before ALD model establishment.Real-time PCR was used to detect miR-203 levelss in liver tissues,and HE staining was used to evaluate the injury of liver tissues.The changes of superoxide dismutase(SOD),catalase(CAT)and malonaldehyde(MDA)in liver tissue homogenates in each group were detected by ELISA and hepatic IL-1βand NF-κB protein expression were detected by Western Blot.Results At the end of eight week model establishment,the miR-203 level in group C rat liver tissue was(2.8±0.1),significantly higher than[(1.3±0.5),P<0.05]in group B;serum AST,ALT and bilirubin levels in group C were(89.5±7.9)U/L,(38.5±10.1)U/L and(27.8±5.1)μmol/L,significantly lower than[(162.8±16.5)U/L,(69.6±7.5)U/L and(54.9±7.8)μmol/L,P<0.05],respectively,in group B;hepatic homogenate SOD and CAT in group C were(57.6±11.4)U/mg and(54.6±9.9)U/mg,significantly higher than[(41.6±7.6)U/mg and(45.7±6.0)U/mg,P<0.05],respectively,in group B,while hepatic homogenate MDA level was(2.8±0.1)nmol/g,much lower than[(5.0±0.2)nmol/g,P<0.05]in group B;the results by Western bloting showed thant hepatic IL-1βand NF-κB expression in group C were(0.7±0.2)and(0.3±0.1),significantly lower than[(1.2±0.3)and(1.0±0.2),respectively,P<0.05]in group B.Conclusion Our findings suggest that the overexpression of miR-203 has a protective effect on liver injury in ALD rats,which might be attributed to the intensified antioxidant and anti-inflammatory effects.