18α-glycyrrhetinic acid extracted from Glycyrrhiza radix inhibits proliferation and promotes apoptosis of the hepatic stellate cell line

Objective: To evaluate the effect of 18α-glycyrrhetinic acid (18α-GA) on the proliferation and Apoptosis of hepatic stellate cells (HSCs) and its underlying mechanisms.

Methods: HSCs (both human and rat HSCs) were pretreated with or without selective peroxisome proliferator-activated receptor-γ (PPAR-γ) antagonist, GW9662, before 18a-GA treatment. Cell cycle and Apoptosis of HSCs were analyzed by flow cytometry, and changes in cell cycle and apoptosis-related proteins were analyzed by Western blot. The effect of 18α-GA on nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) DNA-binding activity was measured by ArrayStar transcription factor activity assay.

Results: 18α-GA markedly reduced LX-2 cell numbers by 14.8% and 31.2% after 48 h and 72 h of treatment, respectively (P < 0.05). 18α-GA also significantly increased the percentage of LX-2 cells in phase G0/G1 and decreased it in phase S after treated for 48 h and 72 h compared with the control group. 18α-GA increased Apoptosis to 6.8% at 48 h, compared with control (2.5%), and at 72 h the percentages of apoptotic cells in control and the treatment groups were 3.1% and 15.6%, respectively, in LX-2 cells (P < 0.01). Similar changes occurred in CCl₄-cirrhotic fat-storing cells. Furthermore, 18α-GA induced expression of PPAR-γ and altered some cell cycle and apoptosis-related proteins. 18α-GA also inhibited NF-κB DNA-binding activity. All these effects were abolished by GW9662.

Conclusions: 18α-GA inhibits the proliferation of activated HSCs and induces Apoptosis in culture. It also increases PPAR-γ expression and decreases NF-κB DNA-binding activity, which may be involved in these effects.