Chinese Hepatolgy ›› 2019, Vol. 24 ›› Issue (6): 638-642.

• Original Articles • Previous Articles     Next Articles

Establishment and evaluation of a fructose-induced hepatic steatosis cell model

He Wenxi, Yang Jinyu, Xu Yanjiao, Lan Lulu, Zhang Chengliang, Liu Dong   

  1. Department of Pharmacy, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
  • Received:2019-01-03 Published:2020-03-30
  • Contact: Zhang Chengliang, Email: ph3719@aliyun.com; Liu Dong, Email:ld2069@outlook.com

Abstract: Objective To establish a L02 cell model of hepatic steatosis induced by fructose, and to study the lipid synthesis of the model.Methods L02 cells were cultured in vitro and treated with different concentrations of fructose for 24 hours to induce steatosis. Cell viability was detected by cell-counting kit-8 to assess the effects of fructose. Levels of alanine aminotransferase and aspartate aminotransferase in the medium were detected to assess the damage of fructose on hepatocytes. Oil red O staining was used to observe the intracellular lipid droplet deposition. The intracellular triglyceride (TG) content was measured to determine the optimal modeling concentration. Meanwhile, the protein levels of carbohydrate responsive element binding protein (ChREBP), sterol regulatory element binding protein-1c (SREBP-1c), acetyl-CoA carboxylase 1 (ACC1) and stearoyl-CoA desaturase 1 (SCD1) in L02 cells treated with fructose were detected and compared with those in cells treated with free fatty acid (FFA). Results There was no significant difference between the viability of L02 cells under the intervention of 0?32 mmol/L fructose, and no significant cell injury was observed. When the fructose concentration was 4 mmol/L, a large amount of lipid droplets were formed in the L02 hepatocytes, the intracellular TG content was significantly higher, which was 1.5 times than that of the normal control, and the intracellular protein levels of ChREBP, SREBP-1 and ACC1 were significantly higher than those of FFA-treated cells.Conclusion The L02 cell steatosis can be successfully induced by 4 mmol/L fructose. This model is suitable for investigating the de novo lipogenesis in non-alcoholic fatty liver disease induced by high fructose diet.

Key words: Fructose, Non-alcoholic fatty liver disease, Cell model, Steatosis, De novo lipogenesis