Investigating the expression of autophagy genes during the in vitro activation of mouse hepatic stellate cells.

Abstract

This study aims to evaluate the expression level of autophagy genes during the activation of mouse hepatic stellate cells (HSCs) in vitro. The HSCs isolated from the mouse liver were cultured in vitro for 7 days. The activation of HSCs was evaluated by their morphology, the storage of lipid droplets by oil red O (ORO) staining, the expression of activation-related genes α-sma, collagen I, and quiescence-related gene lrat by qRT PCR and ICC staining (α-SMA). The expression of autophagy genes lc3b, beclin 1, atg12 were assessed by qRT PCR and ICC staining (LC3). The results showed that the isolated HSCs were activated after 3 days and 7 days of the culture in vitro. The activation was indicated by the morphological change of HSCs to myofibroblast-like cells, loss of lipid droplets, and increased expression of fibrotic genes α-sma and collagen I, decreased expression of lrat. Additionally, the expression of autophagy genes lc3b, beclin 1, and atg12 were significantly increased in the activated HSCs after the culture in vitro for 3 and 7 days. This study contributes the preliminary results to further studies on the role of autophagy during the activation of HSCs, which may be exploited for the development of the antifibrotic therapy targeting autophagy.