Background: Liver fibrosis is a dysregulated wound-healing process in
response to diverse liver injuries, and an effective drug therapy is not yet
available. Genistein, which is one of the most active natural flavonoids mainly
derived from soybean products (e.g., Cordyceps sinensis mycelium),
exhibits various biological effects, including hepatoprotective and
anti-inflammatory properties. However, the anti-hepatic fibrosis mechanisms of
genistein are poorly understood. The aim of our research is to explore the effect
and the possible mechanism of genistein against liver fibrosis. Materials
and methods: Cell counting kit-8, EdU, and flow cytometry assays were applied to
evaluate the effects of genistein on cell viability, proliferation, and cell
cycle arrest in human hepatic stellate cell (HSC) line LX2 cells. HSC activation
was induced by transforming growth factor-1 in LX2 cells and liver
fibrosis model was established by the intraperitoneal injection of
dimethylnitrosamine (DMN) in rats to assess the anti-fibrosis effects of
genistein in vivo and in vitro models. HSC activation was
assessed by qRT-PCR, Western blot, immunohistochemistry, and immunofluorescent
assay. Liver injury and collagen deposition were evaluated by histopathological
assay, serum biochemistry, and hepatic hydroxyproline content assays. The mRNA
expressions of matrix metalloproteinases (MMPs), tissue inhibitors of
metalloproteinases (TIMPs), and inflammation related-factors were assessed by
qRT-PCR assay. Furthermore, the functional properties of macrophage in the liver
were assessed by immunohistochemistry assay. The expression levels of the
JAK2/STAT3/SOCS3 signaling pathway related-protein were assessed by Western blot
analysis. Results: Genistein significantly inhibited cell viability and
proliferation and induced cell cycle arrest at G0/G1 phase in LX2 cells,
respectively. Furthermore, oral administration of genistein significantly
ameliorated liver injury and the collagen deposition in rats with
DMN-induced fibrosis model. Genistein suppressed the expression
levels of HSC activation marker -smooth muscle actin and collagen type
I alpha 1 in vivo and in vitro. Genistein significantly
decreased the mRNA expression levels of extracellular matrix degradation genes
MMP2/9 and TIMP1 in rats. Genistein alleviated the mRNA expression levels of
IL-1, IL-6, TNF-, and MCP-1 and regulated the protein
expressions of CD68, CD163, and CD206 in the liver. Moreover, genistein
attenuated the expressions of p-JAK2/JAK2, p-STAT3/STAT3, and SOCS3 protein both
in vivo and in vitro. Conclusion: Taken together, our
results showed that genistein could be improved liver fibrosis both in
vivo and in vitro, probably through regulating the functional
properties of macrophage and inhibiting the JAK2/STAT3/SOCS3 signaling pathway.