Validation and functional analysis of the critical proteins in combination with taurine, epigallocatechin gallate and genistein against liver fibrosis in rats.

In our previous works, we highlight nine candidates (Cathepsin D, Lamp1, Tpi1, Fgb, FVII, Mst4, CDK4, Hdgf and Glud1) that might be key proteins of combination therapy anti-fibrosis in rats. In this research, in order to verify the function of candidates, gene or protein expression of the nine candidates was verified by Western blot and RT-qPCR, and enzyme-linked immunosorbent assay in vitro and vivo. The expression of Fgb, Tpi1, CDK4, Mst4 and FVII significantly changed and matched with previous results after combination treating fibrosis rats or hepatic stellate cells (HSCs). These proteins may play crucial roles in anti-fibrosis. In particular, FVII take on pivotal role in combination therapy against liver fibrosis. The viability and cycle of HSCs was determined using CCK8 assay and Flow Cytometry, respectively. The results indicate that an overexpression of FVII could accelerate HSC proliferation and reduce the pharmacologic sensitivity. Combination therapy may inhibit HSC proliferation by blocking cell cycle in S phase. Although further studies are necessary to determine the precise protein functions, this research provides the possible molecular mechanisms and signaling pathways of combination therapy against liver fibrosis.