Studying the impact of autophagy inhibition by chloroquine to primary hepatic stellate cell activation in vitro

Abstract

This paper presented the isolation of primary hepatic stellate cell (HSC) to investigate the effects of chloroquine (CQ) on autophagy during the primary hepatic stellate cell activation in vitro. After isolation, HSC cells were evaluated for the isolation efficiency through the cell number and specific characteristics of HSC cells, such as cell size, lipid droplet storage capacity by Oil Reo O (ORO) staining, Desmin expression by Immunocytochemistry (ICC) staining, and morphological changes during HSC culture in vitro. The HSC after being isolated for 1 day were treated with CQ 10 μM for 24 h. The ability of CQ to inhibit the autophagy and prevent HSC activation in vitro was assessed through their morphology and the expression levels of LC3, P62, α-SMA, Collagen I (COL I) by ICC staining. The results showed that CQ 10 μM inhibited the autophagy via increasing the expression of LC3 (43.31 ± 1.61%) and P62 (61.28 ± 2.64%) compared with the control at day 2. CQ 10 μM also prevented HSC the activation in vitro through the incomplete transformation of myofibroblast-like cells and decreased expression of α-SMA (24.17 ± 7.40%) and COL I (18.05 ± 0.49%) at day 7 compared with the control. This result could be one step in determining the relationship between the autophagy and HSC activation, aiming to develop therapy targeting autophagy in the treatment of liver fibrosis.