Lei Lian1,2, Qunsheng Huang3,4, Longjuan Zhang5, Huabo Qin3,6, Xiaosheng He3,4, Xin He3,7, Jia Ke3,4, Minghao Xie3,8, Ping Lan9,10. 1. Department of Colorectal Surgery, The Sixth Affiliated Hospital of Sun Yat-sen University, 26 Yuancun Er Heng Rd., Guangzhou, 510655, Guangdong, China. lianlei2@mail.sysu.edu.cn. 2. Guangdong Institute of Gastroenterology, Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China. lianlei2@mail.sysu.edu.cn. 3. Department of Colorectal Surgery, The Sixth Affiliated Hospital of Sun Yat-sen University, 26 Yuancun Er Heng Rd., Guangzhou, 510655, Guangdong, China. 4. Guangdong Institute of Gastroenterology, Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China. 5. Laboratory of Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China. 6. Department of General and Pediatric Surgery, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, China. 7. Department of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou City, Guangdong, China. 8. Department of General Surgery, The Affiliated Hospital of Jiujiang University, Jiujiang, Jiangxi, China. 9. Department of Colorectal Surgery, The Sixth Affiliated Hospital of Sun Yat-sen University, 26 Yuancun Er Heng Rd., Guangzhou, 510655, Guangdong, China. lanping@mail.sysu.edu.cn. 10. Guangdong Institute of Gastroenterology, Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China. lanping@mail.sysu.edu.cn.
Abstract
BACKGROUND: Intestinal fibrosis is a major complication of CD and may result in stricture formation leading to intestinal obstruction. MSCs play multiple roles in active CD and fibrosis-associated diseases. AIMS: This study was designed to investigate the role of MSCs in CD-associated intestinal fibrosis. METHODS: Intestinal fibrosis was induced over 7 weeks of enema with increasing doses of TNBS and assessed by Masson's trichrome staining. Transcriptome sequencing and gene set enrichment analysis were conducted to reveal the transcriptome changes among groups at the mRNA level. Immunofluorescence assays were used to validate the role of EMT in intestinal fibrosis. Quantitative real-time PCR and immunohistochemistry analyses were performed to clarify the association between the anti-fibrogenic properties of MSCs and the immune microenvironment. Western blotting was used to verify the potential signaling pathways. RESULTS: Fibrotic tissue accumulation and inflammatory cell infiltration were detected in the colon tissue after TNBS induction treatment. Prophylactic MSCs treatment inhibited colon shortening, while therapeutic treatment decreased colon weight. Prophylactic treatment with MSCs inhibited the accumulation of fibrotic tissue, the expression of fibrotic proteins and EMT. Therapeutic MSCs treatment reversed the established intestinal fibrosis and reduced EMT. The secretion of the fibrogenic factors IL-1beta, IL-6 and IL-13 was down-regulated after both MSCs treatment approaches, while IL-10, an anti-fibrogenic factor, was up-regulated. Both MSCs therapies inhibited the expression of TGF-beta and the phosphorylation of Smad2 and Smad3 after TNBS induction. CONCLUSION: MSCs exert anti-fibrogenic activity against CD-associated fibrosis by regulating the inflammatory environment, inhibiting the TGF-beta/Smad signaling pathway and ameliorating EMT.
BACKGROUND:Intestinal fibrosis is a major complication of CD and may result in stricture formation leading to intestinal obstruction. MSCs play multiple roles in active CD and fibrosis-associated diseases. AIMS: This study was designed to investigate the role of MSCs in CD-associated intestinal fibrosis. METHODS:Intestinal fibrosis was induced over 7 weeks of enema with increasing doses of TNBS and assessed by Masson's trichrome staining. Transcriptome sequencing and gene set enrichment analysis were conducted to reveal the transcriptome changes among groups at the mRNA level. Immunofluorescence assays were used to validate the role of EMT in intestinal fibrosis. Quantitative real-time PCR and immunohistochemistry analyses were performed to clarify the association between the anti-fibrogenic properties of MSCs and the immune microenvironment. Western blotting was used to verify the potential signaling pathways. RESULTS: Fibrotic tissue accumulation and inflammatory cell infiltration were detected in the colon tissue after TNBS induction treatment. Prophylactic MSCs treatment inhibited colon shortening, while therapeutic treatment decreased colon weight. Prophylactic treatment with MSCs inhibited the accumulation of fibrotic tissue, the expression of fibrotic proteins and EMT. Therapeutic MSCs treatment reversed the established intestinal fibrosis and reduced EMT. The secretion of the fibrogenic factors IL-1beta, IL-6 and IL-13 was down-regulated after both MSCs treatment approaches, while IL-10, an anti-fibrogenic factor, was up-regulated. Both MSCs therapies inhibited the expression of TGF-beta and the phosphorylation of Smad2 and Smad3 after TNBS induction. CONCLUSION: MSCs exert anti-fibrogenic activity against CD-associated fibrosis by regulating the inflammatory environment, inhibiting the TGF-beta/Smad signaling pathway and ameliorating EMT.
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