Effects of amylin on triglyceride metabolism in steatosis human L-02 hepatocyte

Abstract

Objective To investigate the effects of amylin on triglyceride metabolism in steatosis human L-02 hepatocyte. Methods The normal hepatoctyes were incubated with 50% fetal bovine serum high-glucose Duchenne modified Eagle's medium (DMEM) for 72 h to establish the hepatic steatosis model. The experiments were divided into normal hepatocyte cells (Group A), steatosis hepatocyte cells (Group B), steatosis hepatocyte cells incubated with different concentrations of amylin for 24 h (0.01 μmol/L amylin (Group C), 0.1 μmol/L amylin (Group D), 1 μmol/L amylin (Group E) and 5 μmol/L amylin (Group F)). The oil red O staining was used to observe the hepatocyte morphology and intracellular lipid droplets; the hepatocyte triglyceride quantitative detection kit was used to detect the triglyceride levels in each group (mg/g protein); reverse transcription-polymerase chain reaction was used to examine the synthase mRNA and acetyl coenzyme A carboxylase mRNA expression levels in each group. Comparison between the results of the different groups was performed with either one-way analysis of variance (ANOVA ) or Student's t test. Results There were lots of red lipid droplets in hepatocyte cells after incubated with 50% fetal bovine serum in high-glucose DMEM medium for 72 h ; the triglyceride levels in group A was (415±52) mg/g protein, and the content was significantly increased in group B with (1129±96) mg/g protein (t=874, P<0.05). After incubated with different concentrations of amylin for 24 h, the triglyceride levels in group C, D, E and F were separately(898±77), (740±83), (515±30) and (628± 48) mg/g protein (F=48.0, P<0.05). The red lipid droplets were reduced by oil Red O staining. Reverse transcription-polymerase chain reaction showed that the intracellular fatty acid synthase mRNA and acetyl coenzyme A carboxylase mRNA expression levels were increased in steatosis hepatocytes and reduced after steatosis hepatocytes incubated with different concentrations of amylin. Conclusion When incubated with amylin, triglyceride content in steatosis hepatocytes decreases and the steatosis alleviates and it may be associated with change of the expression levels of fatty acid synthase mRNA and acetyl coenzyme A carboxylase mRNA. Key words: Amylin; Hepatocyte; Triglyceride