Attenuation of early liver fibrosis by herbal compound “Diwu Yanggan” through modulating the balance between epithelial-to-mesenchymal transition and mesenchymal-to-epithelial transition

Abstract

BackgroundDiwu Yanggan (DWYG) is a Chinese compound herbal preparation which consists of five Chinese herbs. This study investigates the preventative effects of DWYG on liver fibrosis induced by carbon tetrachloride (CCl4) and explores its possible mechanisms of action.MethodsLiver fibrosis was induced in male Wistar rats by injecting a 50% CCl4/soybean oil solution subcutaneously twice a week for six weeks. After six weeks of treatment, serum aspartate transaminase (AST) and alanine transaminase (ALT) assay, liver tissue histological assessment and hepatic hydroxyproline assay were respectively carried out to examine the effects of DWYG on liver function and fibrosis degree. The impacts of DWYG on the expression levels of epithelial marker E-cadherin, mesenchymal marker Vimentin, transforming growth factor β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7) were further examined by quantitative real-time RT-PCR and Western blot analysis. In addition, the differences of Hedgehog (Hh) signaling pathway activity between DWYG-treated and DWYG-untreated fibrotic liver tissues were also evaluated by quantitative real-time RT-PCR and Western blot analysis.ResultsUpon DWYG treatment, the serum levels of ALT and AST, hepatic hydroxyproline content and the degree of fibrosis in CCl4-induced fibrotic model rats were dramatically declined. In accompany with the alleviation of the degree of fibrosis, DWYG treatment provoked the reversal of epithelial-to-mesenchymal transition (EMT) to mesenchymal-to-epithelial transition (MET) in the fibrotic liver tissues, which was characterized with the up-regulation expression of E-cadherin and down-regulation expression of Vimentin. Furthermore, we observed that the expression level of TGF-β1 was reduced whereas the expression level of BMP-7 was enhanced in liver tissues of DWYG-treated rats, therefore the expression ratio of TGF-β1/BMP-7 was dramatically decreased compared to CCl4-induced fibrosis model rats. In addition, quantitative real-time RT-PCR and Western blot analysis demonstrated that after DWYG treatment the expressions of Hh ligand Shh, receptor Smo and Ptc, and transcription factor Gli1 in CCl4-induced fibrotic liver tissues were dramatically repressed.ConclusionsDWYG demonstrates therapeutic potential to prevent liver fibrosis by modulating the balance between EMT and MET through reducing the expression ratio of TGF-β1/BMP-7 and inhibiting the excessive activation of Hh signaling pathway.

Forskolin, a hedgehog signalling inhibitor, attenuates carbon tetrachloride‐induced liver fibrosis in rats

“…In agreement with previous studies, our results showed that liver fibrosis induced by CCl 4 is associated with excessive activation of Hh signalling as shown by increased expression of the Hh pathway components, Ptch‐1, Smo and the transcriptional activator Gli‐2, where the level of Hh pathway activation paralleled the increase in hepatic collagen content (Fleig et al , ; Choi et al , ; Shen et al , ). Activation of the Hh pathway results in Smo‐dependent inhibition of PKA.…”

Liver fibrosis staging using T1 mapping on gadoxetic acid-enhanced MRI compared with DW imaging

“…First, it will distort the hepatic architecture and vasculature; second, it will have a deleterious effect on hepatic function; and third, it will increase the propensity for neoplastic transformation. Many studies have showed that liver fibrosis could be reversed, for example, by administering antiviral treatment for chronic viral hepatitis 4,5 . In addition, the severity of liver fibrosis affects the prognosis of liver cancer surgery.…”

Liver regeneration in traditional Chinese medicine: advances and challenges

“…TGF-b1, epithelialmesenchymal transition, hedgehog signaling pathways are important mechanisms affecting HSC proliferation and differentiation [64]. Recent studies found that the TCM compound Diwu Yanggan Capsule is able to inhibit the overexpression of TGF-b1, inhibit the overactivation of hedgehog signaling pathway, prevent the occurrence and development of hepatic fibrosis, and promote liver regeneration and repair by inhibiting the EMT process of HSCs and/ or promoting its MET process [65][66][67].…”