Effects of in vitro lipid overload on LX-2 hepatic stellate cells

Abstract

Introduction and Objectives: Studying lipid overload repercussions on hepatic stellate cells (HSC) is of great importance due to their role in fibrosis during NAFLD. Steatogenic cell culture of HSC is intended to elucidate pathogenic mechanisms in NAFLD. Materials and Methods: LX-2 HSC were cultured in standard DMEM. Steatogenic medium was prepared: mild steatosis (MS:50µM sodium oleate/sodium palmitate (OA/PA) at 2:1 ratio), severe steatosis (SS:500µM 2OA:1PA). Control (C) was cultured in DMEM. Cells were pre-incubated in DMEM at standard conditions for 24h the incubated in MS or SS medium. Cells were incubated for up to 72h. Viability and mortality ratios were assessed; cellular proliferation and senescence were assessed. Data: Mean ± SD, two-way ANOVA followed by Tukey. P<0.05. Results: Cell viability in MS significantly diminished by 13.6% at 72h, whereas SS showed 49.6 % lower viability from 48h compared with C. Regarding mortality rate, it was increased by 16.0% in MS from 72h and by 50.0% in SS from 48h compared with C. Proliferation was increased in both MS and SS at 24h and significantly decreased by 72h compared with C. Cellular senescence in both steatogenic conditions was diminished among 1.8-22.4% compared with C at 24 and 48h. Conclusion: Steatogenic conditions induced an increased proliferation and lower senescence in LX-2 HSC at 24h in both MS and SS groups. These findings suggest that HSC might turn into an activated state. Our results agree with other reports showing that HSC activation and transdifferentiation increase their proliferation, avoiding other cellular processes, including senescence while contributing to the pathogenesis of NAFLD. Funding: This work was funded by CONACYT (CB-221137) Declaration of interest: The authors declare no potential conflicts of interest.