Published online Aug 14, 2018. doi: 10.3748/wjg.v24.i30.3414
Peer-review started: May 10, 2018
First decision: June 11, 2018
Revised: June 27, 2018
Accepted: June 28, 2018
Article in press: June 28, 2018
Published online: August 14, 2018
Processing time: 95 Days and 17.6 Hours
Epithelial-mesenchymal transition (EMT) is a crucial process in Crohn’s disease (CD) intestinal fibrosis. Total flavone of Abelmoschus manihot (TFA) has been found as an effective component to reduce CD intestinal fibrosis in vivo. However, the role and mechanism of TFA on EMT progress of CD intestinal fibrosis have not been understood yet.
EMT is a crucial process in CD intestinal fibrosis. TFA has been found as an effective component to reduce CD intestinal fibrosis in vivo. However, the role and mechanism of TFA on EMT progress of CD intestinal fibrosis have not been understood yet. In the present study, we performed CCK-8, morphology, wound healing, transwell, qRT-PCR, western blotting and immunofluorescence assays to explore the role and the underlying mechanisms of TFA on CD intestinal fibrosis, and the results indicated that TFA was expected to advance as a new therapy to treat CD intestinal fibrosis.
To explore the role and mechanism of TFA on EMT progress of CD intestinal fibrosis.
First, a CCK-8 assay was performed to assess the effect of TFA on the viability of IEC-6 cells and to 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 transforming growth factor-β1 (TGF-β1). In addition, immunofluorescence, 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 was examined by qRT-PCR, western blotting, morphology, wound healing and transwell assays.
In this study, TFA promoted TGF-β1-induced IEC-6 cell 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 TEC-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 alone.
These 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.
TFA promoted the induction of EMT partly by inhibiting TGF-β1-activated Smad signaling pathway and non-Smad signaling pathway. TFA is expected to advance as a new therapy to treat CD intestinal fibrosis, and its continued advancement may open the door to a new class of treatment for CD intestinal fibrosis.