Triptolide exerts protective effects against fibrosis following ileocolonic anastomosis by mechanisms involving the miR-16-1/HSP70 pathway in IL-10-deficient mice.

Abstract

Surgeries, particularly ileocecal resection (ICR), are often required in the treatment of Crohn's disease (CD). However, recurrences are common for patients who undergo ICR, and anastomotic fibrosis is the main cause of re-operation. The present study aimed to investigate the therapeutic effects of triptolide (TPL) in ameliorating fibrosis following ileocolonic anastomosis. A model of IL-10−/− mice undergoing ICR was used to study post surgical inflammation and fibrosis of anastomosis. For this purpsose, interleukin (IL)-10−/− mice were randomly divided into 3 groups as follows: the control group, the saline-treated group subjected to ICR (ST-ICR) and the TPL-treated group subjected to ICR (TT-ICR). Wild-type (WT) mice of matching ages were assigned to the WT group. The effects of TPL treatment on ileocolonic anastomosis were determined by histopathological evaluation, western blot analysis and ELISA. The analysis of the effects of TPL treatment on microRNA-16-1 (miR-16-1) and heat shock protein 70 (HSP70) expression was carried out by RT-qPCR and western blot analysis. Compared with the control group, significantly higher inflammation scores following anastomosis were observed in the ST-ICR group (P<0.05), although reversion was observed in the TT-ICR group, which was consistent with changes in the area of CD4+ cell infiltration. The elevated fibrosis scores and the overexpression of procollagen I and III in the ST-ICR group were all inhibited by TPL. With an increase in the severity of inflammation and fibrosis, the levels of IL-6, tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) increased; however, a significant decrease in these levels was observed following treatment with TPL (P<0.05). The results of RT-qPCR revealed that the upregulated miR-16-1 levels in the ST-ICR group were significantly reduced by TPL. HSP70, which can be inhibited by miR-16-1, ameliorates anastomotic inflammation and fibrosis. Thus, the present study demonstrates that TPL exerts a protective effect against fibrosis following anastomosis in CD. The miR-16-1/HSP70 signaling pathway, which can be regulated by TPL, may thus represent a novel therapeutic option in CD that deserves further investigation.