Total flavone of Abelmoschus manihot suppresses epithelial-mesenchymal transition via interfering transforming growth factor-β1 signaling in Crohn’s disease intestinal fibrosis

Abstract

AIMTo explore the role and mechanism of total flavone of Abelmoschus manihot (TFA) on epithelial-mesenchymal transition (EMT) progress of Crohn’s disease (CD) intestinal fibrosis.METHODSFirst, CCK-8 assay was performed to assess TFA on the viability of intestinal epithelial (IEC-6) cells and select the optimal concentrations of TFA for our further studies. Then cell morphology, wound healing and transwell assays were performed to examine the effect of TFA on morphology, migration and invasion of IEC-6 cells treated with TGF-β1. In addition, immunofluorescence, real-time PCR analysis (qRT-PCR) and western blotting assays were carried out to detect the impact of TFA on EMT progress. Moreover, western blotting assay was performed to evaluate the function of TFA on the Smad and MAPK signaling pathways. Further, the role of co-treatment of TFA and si-Smad or MAPK inhibitors has been examined by qRT-PCR, western blotting, morphology, wound healing and transwell assays.RESULTSIn this study, TFA promoted transforming growth factor-β1 (TGF-β1)-induced (IEC-6) morphological change, migration and invasion, and increased the expression of epithelial markers and reduced the levels of mesenchymal markers, along with the inactivation of Smad and MAPK signaling pathways. Moreover, we revealed that si-Smad and MAPK inhibitors effectively attenuated TGF-β1-induced EMT in IEC-6 cells. Importantly, co-treatment of TFA and si-Smad or MAPK inhibitors had better inhibitory effects on TGF-β1-induced EMT in IEC-6 cells than either one of them.CONCLUSIONThese findings could provide new insight into the molecular mechanisms of TFA on TGF-β1-induced EMT in IEC-6 cells and TFA is expected to advance as a new therapy to treat CD intestinal fibrosis.

Exosomes derived from mesenchymal stem cells reverse EMT via TGF-β1/Smad pathway and promote repair of damaged endometrium

“…Therefore, it is widely used for EMT induction in vitro. EMT has been found in many tissues/cells, such as bronchial epithelial cells [34], renal tubular epithelial cells [35], intestinal epithelial cells [36], and retinal epithelial cells [37]. All of these cells could undergo EMT under the induction of TGF-β1.…”

Total flavone of Abelmoschus Manihot improves colitis by promoting growth of Akkermansia in mice

“…Animal experiments and clinical studies have found that the increase of pro-in ammatory cytokines further damages the intestinal mucosal barrier function through the activation of the NF-κB signaling pathway. Our previous study has shown that TFA could markedly inhibit the release of the intestinal in ammatory cytokines in Crohn's disease rat induced by 2,4,6-trinitrobenzene sulfonic acid, improve intestinal in ammation and ameliorate the colitis symptoms 15 . In this study, we demonstrated supplementation of TFA signi cant decrease the mRNA expression of in ammatory cytokines (TNF-α, IL-6, IL-18, IL-17a, IFN-γ) and chemokines (CCL2) in colon tissue.…”

“…re uxed the mixture for 60 mins at 90℃ then ltered with analytical lter paper. Finally, rotary evaporation was used to evaporate the extracts under vacuum at 60℃ 5 . DSS (molecular weight of 36-50 kDa) was provided by German MP Biopharmaceutical Company.…”

Traditional Uses, Chemical Constituents, Biological Properties, Clinical Settings, and Toxicities of Abelmoschus manihot L.: A Comprehensive Review

“…TFAM at concentrations of 5, 10, and 15 μg/mL effectively suppressed TGF-β1-induced morphological changes, migration, and invasion; increased the mRNA and protein levels of E­cadherin and ZO-1; decreased the mRNA and protein levels of vimentin and N­cadherin; and reduced the levels of Smad 2/3, p-p38, p-c-Jun N-terminal kinase, and p-ERK 1/2 , which involved the Smad/MAPK signaling pathway. In addition, si-Smad and MAPK inhibitors effectively suppressed the TGF-β1-induced epithelial–mesenchymal transition in IEC-6 cells ( Yang B. L. et al., 2018 ). Thus, a combination of TFAM with si-Smad or MAPK inhibitors had greater inhibitory effects on the TGF-β1-induced epithelial–mesenchymal transition in IEC-6 cells, which suggests that TFAM might represent a novel therapeutic strategy for the treatment of intestinal fibrosis in Crohn’s disease.…”

“…Modern pharmacological findings have shown that the extracts and active constituents of A. manihot possess various biological properties, including anti-DN ( Tu et al., 2013 ), anticonvulsant ( Guo et al., 2011 ), antioxidant ( Zhang et al., 2013 ), antiadipogenic, anti-inflammatory ( Jain and Bari, 2010 ), analgesic ( Fan et al., 2003 ), antidepressant ( Guo et al., 2011 ), antiviral ( Wu et al., 2007 ), antitumor ( Zheng et al., 2016 ), antiplatelet ( Guo et al., 2005 ), anti-Crohn’s disease ( Yang B. L. et al., 2018 ), anti-poststroke depression ( Liu et al., 2009 ), proangiogenic ( Zhu et al., 2018 ), cardioprotective ( Lv et al., 2017 ), neuroprotective ( Cheng et al., 2006 ), immunomodulatory ( Pan et al., 2018 ), and hepatoprotective ( Ai et al., 2013 ) properties, and are effective against cerebral infarction, bone loss ( Guo and Chen, 2002 ; Puel et al., 2005 ), etc. Many of these activities are consistent with those of A. manihot in traditional medicine and support traditional usage.…”