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

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.

MiRNA-21 Regulates Multiple Factors in the Pathogenesis of IBD: A Literature Review

“…The effects of a weakened epithelial barrier also extend to the mucosal barrier of the gut, seen in a study by Zhang et al which explored the Sishen Pill (SSP) which maintained the mucosal barrier of the colon by upregulating PTEN and subsequently downregulating the PI3K/Akt pathway [69]. Additionally, this study illustrates how PTEN downregulates of the Rho A/Rho kinase (ROCK) pathway which when stimulated decreases the integrity of the mucosal barrier.…”

Sishen Pill and its active phytochemicals in treating inflammatory bowel disease and colon cancer: an overview

“…Another study found that when the intestinal vascular barrier is damaged, intestinal bacteria are more likely to spread to the liver, promoting the formation of a pre-metastatic niche for “colon-liver” metastasis, thereby promoting the recruitment of metastatic cells ( Bertocchi et al, 2021 ). As mentioned earlier, Sishen Pills ( Zhang et al, 2021c ), schisandrin B ( Li et al, 2019b ), schisandrin C ( Kim et al, 2022 ), and corylin ( Wang et al, 2023f ) can regulate the secretion of intestinal epithelial tight junction proteins and mucin, repair damaged intestinal mucosal barriers and inhibit the progression of IBD ( Figure 3D ).…”

“…Briefly, it can 1) act on mammalian rapamycin target protein complex 1 (mTORC1) to promote protein synthesis and cell growth; 2) phosphorylate forkhead box O (FOXO) transcription factors, inhibit its transcriptional activity, and affect cell cycle, apoptosis, and metabolism; 3) inhibit the activity of glycogen synthase kinase 3 (GSK3) and regulate glycogen synthesis and cell cycle; 4) phosphorylate and activate the pro-apoptotic protein Bad, making it unable to bind to Bcl-2 or Bcl-XL, and thus reducing the occurrence of normal cell apoptosis ( Mayer and Arteaga, 2016 ; Wang et al, 2023e ). In the above study, Sishen Pill improved intestinal inflammatory factors, immune cell disorders, and a series of symptoms of IBD by downregulating the expression of key proteins in the MAPK ( Zhao et al, 2013 ) and PI3K/Akt ( Ge et al, 2020 ; Zhang et al, 2021c ; Liu et al, 2021 ; Liu et al, 2022 ) signaling pathways, and its active metabolites bavachin ( Wang et al, 2023a ), myristicin ( Duan et al, 2020 ), evodiamine ( Chien et al, 2014 ), schisandrin B ( Jiang et al, 2015 ), 8-methoxypsoralen ( Bartnik et al, 2017 ), and schisandrin B ( Dai et al, 2018 ). Consequently, the Sishen Pill could inhibit the progression of colon cancer by regulating the MAPK and PI3K pathways.…”

“…Wang ( Wang et al, 2019a ) and Ge ( Ge et al, 2022 ) confirmed that TLR4 was the key target, and downregulating TLR4 could inhibit the occurrence of subsequent inflammatory responses through MyD88-dependent and MyD88-independent pathways. In addition, Zhang ( Zhang et al, 2021c ), Wang ( Wang et al, 2022a ) and Zhao ( Zhao et al, 2013 ) explored the molecular mechanism of the Sishen Pill in inhibiting the inflammatory response and promoting intestinal mucosal repair via phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB/Akt), Janus kinase (JAK)/signal transducer and activator of transcription 5 (STAT5), and mitogen activated protein kinase (MAPK) signal pathways. Moreover, the regulation of intestinal immune cells by Sishen Pill mainly manifests in different subsets of T lymphocytes and regulatory T cells (Treg) ( Liu et al, 2016 ), helper T cells (Th) ( Liu et al, 2016 ), follicular helper T cells (Tfh) ( Liu et al, 2020 ), follicular regulatory T cells (Tfr) ( Huang et al, 2022 ; Kang et al, 2022 ), memory T cells (TM) ( Ge et al, 2020 ), and dendritic cells ( Liu et al, 2022 ).…”

“…Researchers found that the richness and diversity of fecal microbiota in postoperative colon cancer patients were lower compared to healthy individuals; the Sishen Pill could improve this trend (Tan et al, 2023). For patients undergoing radical resection of colorectal cancer, Sishen Pill could not only alleviate clinical symptoms such as abdominal distension, tiredness, knee pain, waist acid, and cold but also reduce the tumor marker CEA and immune function indexes CD8 + and CD8 + /CD4 + and increase the level of CD4 + (Zhang Y. et al, 2021). It should be pointed out that all three clinical studies mentioned above exist some methodological flaws, for example, not using a double-blind study design, and lacking relevant descriptions about allocation concealment, which to some extent reduces the reliability of clinical trial results; besides, these studies have not adopted the core indicators of clinical research on colon cancer, such as whether it can improve the survival rate of patients?…”