【摘要】 目的评估干扰miR-135a是否对非酒精性脂肪肝(nonalcoholic fatty liver disease, NAFLD)具有治疗作用。方法采集NAFLD患者以及健康志愿者的血清样本,用qRT-PCR检测miR-135a表达水平;用25、100和400μmol/L棕榈酸(palmitic acid, PA)处理人源原代肝细胞48h,RT-qPCR检测细胞中miR-135a表达水平;将miR-135a inhibitor转入人源原代肝细胞后再给予400μmol/L PA处理48h,尼罗红染色法检测细胞中脂质蓄积情况。通过高脂饮食(high fatty diet, HFD)诱导法制作NAFLD小鼠模型,并在HFD喂养的第7周通过尾静脉注射LV-anti-miR-135a;在第10周末,处死小鼠并收集肝组织和血液样本;肝组织病理学染色用于分析脂质含量及脂肪变性程度,血液生化检测用于分析肝功能和脂质代谢。结果在NAFLD患者血清、PA处理的肝细胞、HFD喂养的小鼠血清和肝组织中miR-135a的表达明显升高;干扰miR-135a能抑制PA诱导的肝细胞中脂质累积;肝组织病理学检测显示,干扰miR-135a能减轻HFD诱导的肝组织脂肪变性;血液生化检测结果显示,干扰miR-135a能降低HFD喂养的小鼠血清中甘油三酯、胆固醇、谷草转氨酶(glutamic-oxaloacetic transaminase,AST)、谷丙转氨酶(alanine transaminase, ALT)、碱性磷酸酶(alkaline phosphatase, ALP)以及总胆红素(total bilirubin,TBIL)的水平。结论干扰miR-135a能够减轻NAFLD引起的肝脂肪变性,并恢复肝功能和改善脂质代谢,因此,抑制体内miR-135a表达水平可能是潜在的NAFLD治疗的新途径之一。更多还原

【Abstract】 Objective To evaluate whether interfering miR-135a has a therapeutic effect on nonalcoholic fatty liver disease(NAFLD).Methods Serum samples from NAFLD patients and healthy volunteers were collected,and the expression level of miR-135a was detected by RT-qPCR.Human primary liver cells were treated with 25,100,and 400μmol/L palmitic acid (PA) for 48h,and the expression level of miR-135a in the cells was detected by qRT-PCR.miR-135a inhibitor was transfected into human primary liver cells and then treated with 400μmol/L PA for 48h,the lipid accumulation in the cells was detected by Nile red staining.NAFLD mouse model was established by high fatty diet (HFD) induction method,and LV-antimiR-135a was injected through tail vein at the 7th week of HFD feeding.At the end of the 10th week,the mice were sacrificed for collecting liver tissues and blood samples.Liver histopathological staining was used to analyze the lipid content and the degree of steatosis,and blood biochemical tests were used to analyze the liver function and lipid metabolism.Results The expression of miR-135a was significantly increased in serum of NAFLD patients,liver cells treated with PA,serum and liver tissues of HFD-fed mice.Interfering miR-135a inhibited PA-induced lipid accumulation in liver cells.Hepatic histopathological results showed that interfering miR-135a alleviated HFD-induced hepatic steatosis.Blood biochemical tests showed that interfering miR-135a could reduce the levels of triglyceride,cholesterol,glutamic-oxaloacetic transaminase (AST),alanine transaminase (ALT),alkaline phosphatase (ALP),and total bilirubin (TBIL) in the serum of HFD-fed mice.Conclusion Interfering miR-135a can alleviate hepatic steatosis induced by NAFLD,restore liver function and improve lipid metabolism.Therefore inhibiting the expression of miR-135a might be one of the potential ways to treat NAFLD.更多还原