摘要
Abstract
Objective To observe the effect of acyl-CoA carbrolase (ACC) antisense oligonucleotide (ASODN) on steatosis of HL-7702 hepatocytes.Methods The specific phosphorothioate modified ACC targeting ASODN were designed and synthesized by a computer software.Normal HL-7702 hepatocytes were cultured in vitro using intravenous lipid emulsions,a large number of triglyceride was induced in hepatocytes within 48 h,and a model of fatty liver cells in vitro was successfully formed.ACC-ASODN was transfected into the hepatocyte model by liposome method.The blank group (HL-7702 cells),model group (fatty liver cells of model) and low and high concentrations of the two experimental groups (5,10 μmol · L-1 ACC-ASODN) were set up.ACC-ASODN was transfected into fatty liver cell model 24 h later.ACC activity was determined by radioisotope technology.The malonyl coenzyme A content was determined by HPLC.The carnitine palmitoyltransferase Ⅰ (CPT Ⅰ) activity was determined by radioisotope technique.The level of triacylglycerol(TG) was determined by automatic analyzer.Results The value of the liver cell ACC activity (U) in blank group,model group and experimental-L,-H groups were respectively 0.74,0.33,0.20,0.18.The value of the intracellular content (nmol · L-1) of malonyl CoA in above four groups were respectively 14.3,12.4,7.5,6.8.The value of CPT Ⅰ (U) in above four groups were respectively 1.33,1.47,3.85,3.95;the value of TG conten(mmol · L-1) in above four groups were respectively 0.20,0.52,0.37,0.28.Compared with blank group,the factors in model group had significant differences(all P ＜ 0.05).Compared with model group,the factors in experimental-L,-H groups had significant differences (all P ＜ 0.01).Conclusion ACC-ASODN acts on hepatocytes,and by inhibiting the activity of ACC,the content of TG is decreased,and the steatosis of hepatocytes is alleviated.关键词
乙酰辅酶A羧化酶/反义寡核苷酸/非酒精性脂肪肝/丙二酰辅酶A/肉碱棕榈酰转移酶ⅠKey words
acetyl coenzyme A carboxylase/antisense oligonucleotide/non-alcoholic fatty liver/malonyl CoA/carnitine palmitoyltransferase Ⅰ分类
医药卫生
