Mechanism of acacetin regulating hepatic stellate cell apoptosis based on network pharmacology and experimental verification

Background: Hepatic fibrosis is caused by various liver diseases and eventually develops into liver Cancer. There is no specific drug approved for the treatment of hepatic fibrosis in the world. Acacetin (AC), a natural flavonoid, is widely present in nature in various Plants, such as black locust, Damiana, Silver birch. It has been reported that acacetin can inhibit the proliferation of Cancer cells and induce Apoptosis.

Purpose: In this study, we investigated the effect of acacetin on hepatic stellate cell Apoptosis, thereby improving hepatic fibrosis, and combined experimental validation and molecular docking to reveal the underlying mechanism.

Result: First, we discovered that acacetin inhibited hepatic stellate cell proliferation as well as the expression of fibrosis-related proteins α-smooth muscle actin (α-SMA) and Collagen type I 1 gene (COL1A1) in LX2 cells. Acacetin was then found to promote Apoptosis of hepatic stellate cells through the Caspase cascade pathway. Network pharmacology screening showed that TP53, CASP3, CASP8, BCL2, PARP1, and Bax were the most important targets related to Apoptosis in the PPI network. GO and KEGG analyses of these six important targets were performed, and the top 10 enriched biological processes and related signaling pathways were revealed. Further network pharmacology analysis proved that Apoptosis was involved in the biological process of acacetin's action against hepatic stellate cells. Finally, molecular docking revealed that acacetin binds to the active sites of six apoptotic targets. In vitro experiments further confirmed that acacetin could promote the Apoptosis of LX2 cells by inducing the activation of P53, thereby improving hepatic fibrosis.

Conclusion: acacetin induces P53 activation and promotes Apoptosis of hepatic stellate cells thereby ameliorating hepatic fibrosis.

Acacetin1; Apoptosis2; Hepatic fibrosis3; Molecular Docking4; Network Pharmacology5.