Sishen Pill Maintained Colonic Mucosal Barrier Integrity to Treat Ulcerative Colitis via Rho/ROCK Signaling Pathway.

Xiao-Yun Zhang,Hai-Mei Zhao,Yi Liu,Qi-Hong Pan,Xiu-Yun Lu,Duan-Yong Liu,Yan-Zhen Li,Hai-Yan Wang,Wei Ge,Fatima Martel

doi:10.1155/2021/5536679

Abstract

Sishen Pill (SSP) is a classical prescription of traditional Chinese medicine and often used to treat gastrointestinal diseases, including ulcerative colitis (UC). However, its mechanism is still unclear. We aimed to determine the mechanism of SSP in the treatment of UC by investigating if it maintains the integrity of the intestinal mucosal barrier via the Rho A/Rho kinase (ROCK) signaling pathway. Administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS) successfully induced chronic UC in rats, while the treatment effect of SSP was evaluated by body weight change, colonic length, colonic weight, colonic weight index, histological injury score, and pathological injury score after colitis rats were treated for 7 days. TNF-α and IL-1β levels were analyzed by ELISA, and the proteins of PI3K/Akt and RhoA/ROCK signaling pathway and junction proteins expression were measured by western blotting assay, and the distribution of Claudin 5 was shown by immunofluorescence. SSP significantly improved the clinical symptoms of colitis in rats and reduced the expression of p-RhoA, ROCK1, PI3K, and Akt in the colon mucosa, while it increased the expression of p-Rac and related proteins (Claudin-5, JAM1, VE-cadherin, and Connexin 43). In addition, SSP increased p-AMPKα and PTEN proteins expression, decreased Notch1 level, and hinted that activation of the PI3K/Akt signaling pathway was inhibited. In conclusion, SSP effectively treated chronic colitis induced by TNBS, which may have been achieved by inhibiting PI3K/Akt signal to suppress activation of the Rho/ROCK signaling pathway to finally maintain the integrity of the intestinal mucosal barrier.

Highlights

Ulcerative colitis (UC) is a nonspecific intestinal inflammatory disease
Clinical observations showed that rats in the trinitrobenzene sulfonic acid (TNBS) group curled up had decreased appetite, thin stools, and even hematochezia, with decreased body weight. ese results were consistent with the clinical score (Figure 1(d)). e colonic mucosal damage index (CMDI) score reflects colonic mucosal injury by the naked eye. e CMDI score in the TNBS group was significantly higher than that in the normal group, while the CMDI scores (Figure 1(f)) in the TNBS + Sishen Pill IBD (SSP) and TNBS + 5-ASA groups were lower than that in the TNBS group. e body weight, colon weight, and colon weight index in rats are important indices and reflect the severity of UC
Compared with the normal group, the colon in rats (Figures 1(e) and 1(g)) in the TNBS group was significantly shorter, and following SSP and 5-ASA treatment, colon length was restored (Figures 1(e) and 1(g)) (P < 0.01). ese results demonstrated that the symptoms of experimental colitis induced by TNBS were effectively alleviated in the TNBS + SSP group and the TNBS + 5-ASA group

Summary

Introduction

Ulcerative colitis (UC) is a nonspecific intestinal inflammatory disease. At present, most treatment methods have failed to reduce the incidence of UC. e specific etiology of UC is unclear but is mainly related to genetic susceptibility, immune response disorders, and epithelial barrier damage [1]. An increasing number of studies have reported that the decline of barrier function in intestinal epithelial cells is positively correlated with the degree of mucosal inflammation in UC patients [2, 3]. Intestinal epithelial intercellular connections are the main components of the intestinal mechanical barrier [4], which is mainly composed of tight junctions (TJs) and the cytoskeleton system. When the intestinal epithelial junction is destroyed or the cytoskeleton is damaged, the mucosal barrier integrity of intestinal epithelial will be destroyed to induce ulcer formation. Rho A/ROCK plays an important role in maintaining the connection between intestinal epithelial cells. It can regulate reconstruction of the F-actin skeleton. Many studies showed that Rho/ROCK can affect the cytoskeleton and its close

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