Chen Juan,Shen Wei.Effect of hepatitis B virus X protein on sodium oleate-induced lipid accumulation in HepG2 cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(03):196-200.
Chen Juan,Shen Wei.Effect of hepatitis B virus X protein on sodium oleate-induced lipid accumulation in HepG2 cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(03):196-200.DOI:
Effect of hepatitis B virus X protein on sodium oleate-induced lipid accumulation in HepG2 cells
Western blot检测HBx、LXRα及FAS蛋白表达水平。 结果 转染后16 h HepG2-HBx细胞和HepG2- pIRES2细胞中开始有GFP的表达,提示转染成功;仅在HepG2-HBx细胞内有HBx表达,表明HepG2-HBx细胞模型构建成功。在相同油酸钠处理的条件下,HBx/OA组细胞内脂质含量和TG含量与对照组相比均明显增加(P0.01)。在油酸钠处理细胞24 h,HBx/OA组细胞内LXRα、SREBP-1的mRNA表达量和LXRα、FAS蛋白表达量较对照组均明显增加(P0.01/0.05)。 结论 HBx通过上调HBx-LXRα-SREBP1/FAS通路脂质合成相关基因表达,可能增加HepG2-HBx细胞对外界脂代谢紊乱因素的易感性,从而增加油酸钠诱导HepG2细胞脂质沉积。
Abstract
Objective To investigate whether hepatitis B virus X protein (HBx) increases sodium oleate-induced lipid accumulation in HepG2 cells. Methods HBX gene eukaryotic expression vector pIRES2-EGFP-HBx was transiently transfected into HepG2 cells to establish HepG2-HBx cell model for HBx expression. HepG2 cells transfected with empty vector pIRES2-EGFP (HepG2-pIRES2) and untransfected HepG2 cells (HepG2) were used as controls. Expression of green fluorescent protein (GFP) was observed by fluorescence microscope after transfection. At 16 h after transfection
each group of cells was treated with sodium oleate for 24 and 48 h (HBx/OA group
empty/OA group and G2/OA group
respectively). Lipid accumulation in each group of cells was observed by triglyceride (TG) content detection and oil red O staining. At 24 h after sodium oleate treatment
RT-PCR and Western blot were applied to measure mRNA levels of sterol regulatory element binding protein-1 (SREBP-1) and liver X receptor alpha (LXRα) and protein levels of HBx
LXRα and fatty acid synthase (FAS)
respectively. Results At 16 h after transfection
GFP expression was detected in HepG2-HBx and HepG2-pIRES2 cells
while HBx expression was only detected in HepG2-HBx cells. These suggested that HepG2-HBx cell model for HBx expression was successfully obtained. Compared with those of the control groups
TG and lipid contents were significantly increased in HBx/OA group (P0.01) after the cells treated with sodium oleate. At 24 h after sodium oleate treatment
mRNA levels of LXRα and SREBP-1 as well as protein levels of LXRα and FAS in the HBx/OA group were significantly higher than those in the control groups (P0.01/0.05). Conclusion HBx increases the susceptibility of HepG2-HBx cells to factors causing lipid metabolism disorder through up-regulating expressions of lipid synthesis-related genes of HBx-LXRα-SREBP-1/FAS pathway
thereby increasing sodium oleate-induced lipid accumulation in HepG2 cells.