Objective      To construct miRNA eukaryotic expression vectors targeting SREBP-1c and determine the effect of SREBP-1c gene knockdown on lipid accumulation in L02 and HepG2 hepatocytes under endoplasmic reticulum stress.       Methods      Three miRNA targeting SREBP-1c gene

named SREBP-1c-1-miRNA

SREBP-1c-2-miRNA and SREBP-1c-3-miRNA

and a non-

homologous negative control expression vector (HK-miRNA) were designed and constructed which were identified by DNA sequencing analysis. After 3 miRNA expression vectors were transiently transfected into hepatocytes

fluorescence microscopy was applied to observe EGFP to confirm the transfection. Western blotting was used to determine expression of SREBP-1c. Lipid deposition in each group was observed by triglyceride content detection and oil red O staining. Western blotting was also applied to measure the protein levels of fatty acid synthase (FAS) and acetyl CoA carboxylase (ACC1).       Results      Four miRNA expression vectors were constructed successfully. After 24 hours’ transfection

EGFP expression was visible under fluorescence microscope. The protein level of SREBP-1c was decreased significantly after transfected with SREBP-1c-1miRNA and SREBP-1c-3miRNA

when compared with control group (

P

<

0.05). SREBP-1c-1miRNA was identified with most powerful interference (

P

<

0.05). Compared with the control group

TG content (

P

<

0.05) and lipid droplet were significantly decreased

and FAS and ACC1 proteins were also inhibited in SREBP-1c inference group (

P

<

0.05).       Conclusion      SREBP-1c gene interference suppresses lipid synthesis through down-regulating FAS and ACC1 in L02 and HepG2 hepatocytes under endoplasmic reticulum stress.