Inhibition effect of phytoestrogen calycosin on TGF-β₁-induced hepatic stellate cell activation, proliferation, and migration via estrogen receptor β

Abstract

The present study was designed to investigate the effects of calycosin on hepatic stellate cell (HSC) function and to explore whether the drug exerts its effect through the estrogen receptor. HSC proliferation and migration were measured by MTT assay and transwell chamber assay, respectively. The mRNA and protein expression of α-SMA, COL-I, and ERβ were detected by real-time PCR and Western blotting. The co-localization and expression of α-SMA and ERβ protein were detected by immunofluorescence. All the studies were investigated in the absence or presence of ICI 182,780. The results showed that calycosin inhibited the proliferation of activated HSCs and remarkably inhibited HSC migration. Calycosin significantly reduced the expression of α-SMA and COL-I in activated HSCs. However, with co-treatment with ICI 182,780, the inhibitory effect of calycosin against the above effects was strongly negated. Importantly, calycosin significantly downregulated the expression of ERβ protein, while co-treatment with ICI 182,780 partially reversed the ERβ downregulation. In addition, α-SMA decreased with the decrease of ERβ expression and the subtype of ERβ on HSC is ERβ5. In conclusion, calycosin inhibits proliferation, activation, and migration of TGF-β1-induced HSCs. The effect may be related to binding and downregulation of ERβ5.

Saikosaponin d Alleviates Liver Fibrosis by Negatively Regulating the ROS/NLRP3 Inflammasome Through Activating the ERβ Pathway

“…Soybean isoflavones can alleviate DSS-induced colitis by inhibiting the ERα pathway and downregulating the subsequent activation of the NLRP3 inflammasome ( Gao et al, 2020 ). Phytoestrogen calycosin inhibits the activation, proliferation, and migration of HSCs induced by TGF-β1 via downregulating ERβ ( Deng et al, 2018 ). Comprehensive studies have shown that the mechanism of interaction between estrogen regulators and ligands is complex, and a specific regulator may even have opposite effects under different circumstances.…”

Overexpression of estrogen receptor β inhibits cellular functions of human hepatic stellate cells and promotes the anti-fibrosis effect of calycosin via inhibiting STAT3 phosphorylation

“…According to our previous method [16], the migration ability of LX-2 and transfected cells was inspected by 24-well corning transwell chambers (8.0 μm pore size polycarbonate membrane, Corning Costar, USA). Cells were washed with PBS after digestion and resuspended in serum-free DMEM.…”

“…As phytoestrogens, calycosin inhibits the growth of various tumors through ERβ-mediated signaling pathway. Our previous studies revealed the anti-liver fibrosis effect of calycosin in CCl 4 -induced liver fibrosis in mice and its inhibition effect on the proliferation, migration and activation of HSC-T6 cells [15,16]. JAK/STAT is a key signaling pathway in a variety of cellular activities, including proliferation, differentiation, migration, apoptosis and cell communication, as well as complex biological processes such as inflammation, immune response and cancer development [17].…”

Beneficial and Deleterious Effects of Female Sex Hormones, Oral Contraceptives, and Phytoestrogens by Immunomodulation on the Liver

“…Regarding the PE effects on liver fibrosis by immunomodulation, the O-methylated isoflavone calycosin inhibits the expression of ECM proteins and the proliferation, activation, and migration of HSC induced by TGF-β1. This effect was correlated with calycosin binding and down-regulation of the ERβ5 subtype [195]. Genistein has also been shown to possess hepatoprotective effects in D-GalN-induced fulminant hepatic failure in rats by modulating the NF-κB/MAPK pathways [161] and anti-fibrotic effects in chronic damage by the Smad7-induced inhibition of TGF-β/Smad2/3 [196].…”