Saikosaponin D Inhibits the Proliferation and Promotes the Apoptosis of Rat Hepatic Stellate Cells by Inducing Autophagosome Formation

Objective: This study aimed to investigate the effects of saikosaponin D (SSd) on the proliferation and Apoptosis of the HSC-T6 hepatic stellate cell line and determine the key pathway that mediates SSd's function.

Methods: Cell viability was detected using the CCK-8 kit. The EdU kit and flow cytometry were used to examine cell proliferation. The Annexin V-FITC/PI double staining kit and flow cytometry were used to examine cell Apoptosis. Western blot analysis was performed to analyze the expression levels of LC3, Ki67, cleaved Caspase 3, Bax, and Bcl2. Autophagosome formation was detected by LC3-GFP adenovirus transfection.

Results: SSd inhibits the proliferation and promotes the Apoptosis of acetaldehyde-activated HSC-T6 cells. SSd treatment increased the expression of cleaved Caspase 3 and Bax but reduced that of Ki67 and Bcl2. The same concentration of SSd barely influenced the growth of normal rat liver BRL-3A cells. SSd upregulated LC3-II expression and induced autophagosome formation. Autophagy agonist rapamycin had the same effect as SSd and Autophagy inhibitor 3-methyladenine could neutralize the effect of SSd in acetaldehyde-activated HSC-T6 cells.

Conclusions: SSd could inhibit the proliferation and promote the Apoptosis of HSC-T6 cells by inducing autophagosome formation.