Colonic TRPV4 overexpression is related to constipation severity
Abstract Background Chronic constipation is prevalent and involves both colon sensitivity and various changes in intestinal bacteria, particularly mucosa-associated microflora. Here we examined regulatory mechanisms of TRPV4 expression by co-culturing colon epithelial cell lines with intestinal bact...
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| Format: | Article |
| Language: | English |
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BMC
2023-01-01
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| Series: | BMC Gastroenterology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12876-023-02647-0 |
| _version_ | 1797952499043270656 |
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| author | Hiroshi Mihara Kunitoshi Uchida Yoshiyuki Watanabe Sohachi Nanjo Miho Sakumura Iori Motoo Takayuki Ando Masami Minemura Jibran Sualeh Muhammad Hiroyuki Yamamoto Fumio Itoh Ichiro Yasuda |
| author_facet | Hiroshi Mihara Kunitoshi Uchida Yoshiyuki Watanabe Sohachi Nanjo Miho Sakumura Iori Motoo Takayuki Ando Masami Minemura Jibran Sualeh Muhammad Hiroyuki Yamamoto Fumio Itoh Ichiro Yasuda |
| author_sort | Hiroshi Mihara |
| collection | DOAJ |
| description | Abstract Background Chronic constipation is prevalent and involves both colon sensitivity and various changes in intestinal bacteria, particularly mucosa-associated microflora. Here we examined regulatory mechanisms of TRPV4 expression by co-culturing colon epithelial cell lines with intestinal bacteria and their derivatives. We also investigated TRPV4 expression in colon epithelium from patients with constipation. Methods Colon epithelial cell lines were co-cultured with various enterobacteria (bacterial components and supernatant), folate, LPS, or short chain fatty acids. TRPV4 expression levels and promoter DNA methylation were assessed using pyrosequencing, and microarray network analysis. For human samples, correlation coefficients were calculated and multiple regression analyses were used to examine the association between clinical background, rectal TRPV4 expression level and mucosa-associated microbiota. Results Co-culture of CCD841 cells with P. acnes, C. perfringens, or S. aureus transiently decreased TRPV4 expression but did not induce methylation. Co-culture with clinical isolates and standard strains of K. oxytoca, E. faecalis, or E. coli increased TRPV4 expression in CCD841 cells, and TRPV4 and TNF-alpha expression were increased by E. coli culture supernatants but not bacterial components. Although folate, LPS, IL-6, TNF-alpha, or SCFAs alone did not alter TRPV4 expression, TRPV4 expression following exposure to E. coli culture supernatants was inhibited by butyrate or TNF-alphaR1 inhibitor and increased by p38 inhibitor. Microarray network analysis showed activation of TNF-alpha, cytokines, and NOD signaling. TRPV4 expression was higher in constipated patients from the terminal ileum to the colorectum, and multiple regression analyses showed that low stool frequency, frequency of defecation aids, and duration were associated with TRPV4 expression. Meanwhile, incomplete defecation, time required to defecate, and number of defecation failures per 24 h were associated with increased E. faecalis frequency. Conclusions Colon epithelium cells had increased TRPV4 expression upon co-culture with K. oxytoca, E. faecalis, or E. coli supernatants, as well as TNFα-stimulated TNFαR1 expression via a pathway other than p38. Butyrate treatment suppressed this increase. Epithelial TRPV4 expression was increased in constipated patients, suggesting that TRPV4 together with increased frequency of E. faecalis may be involved in the pathogenesis of various constipation symptoms. |
| first_indexed | 2024-04-10T22:47:19Z |
| format | Article |
| id | doaj.art-9a6a3ea4197a4a78b6301f22cf8156df |
| institution | Directory Open Access Journal |
| issn | 1471-230X |
| language | English |
| last_indexed | 2024-04-10T22:47:19Z |
| publishDate | 2023-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Gastroenterology |
| spelling | doaj.art-9a6a3ea4197a4a78b6301f22cf8156df2023-01-15T12:13:34ZengBMCBMC Gastroenterology1471-230X2023-01-0123111210.1186/s12876-023-02647-0Colonic TRPV4 overexpression is related to constipation severityHiroshi Mihara0Kunitoshi Uchida1Yoshiyuki Watanabe2Sohachi Nanjo3Miho Sakumura4Iori Motoo5Takayuki Ando6Masami Minemura7Jibran Sualeh Muhammad8Hiroyuki Yamamoto9Fumio Itoh10Ichiro Yasuda11Center for Medical Education and Career Development, Graduate School of Medicine and Pharmaceutical Sciences, University of ToyamaDepartment of Physiological Science and Molecular Biology, Fukuoka Dental CollegeDepartment of Internal Medicine, Kawasaki Rinko General HospitalDepartment of Gastroenterology, Graduate School of Medicine and Pharmaceutical Sciences, University of ToyamaDepartment of Gastroenterology, Graduate School of Medicine and Pharmaceutical Sciences, University of ToyamaDepartment of Gastroenterology, Graduate School of Medicine and Pharmaceutical Sciences, University of ToyamaDepartment of Gastroenterology, Graduate School of Medicine and Pharmaceutical Sciences, University of ToyamaDepartment of Gastroenterology, Graduate School of Medicine and Pharmaceutical Sciences, University of ToyamaDepartment of Basic Medical Sciences, College of Medicine, University of SharjahDepartment of Bioinformatics, St. Marianna University Graduate School of MedicineDivision of Gastroenterology and Hepatology, Department of Internal Medicine, St. Marianna University School of MedicineDepartment of Gastroenterology, Graduate School of Medicine and Pharmaceutical Sciences, University of ToyamaAbstract Background Chronic constipation is prevalent and involves both colon sensitivity and various changes in intestinal bacteria, particularly mucosa-associated microflora. Here we examined regulatory mechanisms of TRPV4 expression by co-culturing colon epithelial cell lines with intestinal bacteria and their derivatives. We also investigated TRPV4 expression in colon epithelium from patients with constipation. Methods Colon epithelial cell lines were co-cultured with various enterobacteria (bacterial components and supernatant), folate, LPS, or short chain fatty acids. TRPV4 expression levels and promoter DNA methylation were assessed using pyrosequencing, and microarray network analysis. For human samples, correlation coefficients were calculated and multiple regression analyses were used to examine the association between clinical background, rectal TRPV4 expression level and mucosa-associated microbiota. Results Co-culture of CCD841 cells with P. acnes, C. perfringens, or S. aureus transiently decreased TRPV4 expression but did not induce methylation. Co-culture with clinical isolates and standard strains of K. oxytoca, E. faecalis, or E. coli increased TRPV4 expression in CCD841 cells, and TRPV4 and TNF-alpha expression were increased by E. coli culture supernatants but not bacterial components. Although folate, LPS, IL-6, TNF-alpha, or SCFAs alone did not alter TRPV4 expression, TRPV4 expression following exposure to E. coli culture supernatants was inhibited by butyrate or TNF-alphaR1 inhibitor and increased by p38 inhibitor. Microarray network analysis showed activation of TNF-alpha, cytokines, and NOD signaling. TRPV4 expression was higher in constipated patients from the terminal ileum to the colorectum, and multiple regression analyses showed that low stool frequency, frequency of defecation aids, and duration were associated with TRPV4 expression. Meanwhile, incomplete defecation, time required to defecate, and number of defecation failures per 24 h were associated with increased E. faecalis frequency. Conclusions Colon epithelium cells had increased TRPV4 expression upon co-culture with K. oxytoca, E. faecalis, or E. coli supernatants, as well as TNFα-stimulated TNFαR1 expression via a pathway other than p38. Butyrate treatment suppressed this increase. Epithelial TRPV4 expression was increased in constipated patients, suggesting that TRPV4 together with increased frequency of E. faecalis may be involved in the pathogenesis of various constipation symptoms.https://doi.org/10.1186/s12876-023-02647-0ColonTRPV4E. faecalisE. coliChronic constipation |
| spellingShingle | Hiroshi Mihara Kunitoshi Uchida Yoshiyuki Watanabe Sohachi Nanjo Miho Sakumura Iori Motoo Takayuki Ando Masami Minemura Jibran Sualeh Muhammad Hiroyuki Yamamoto Fumio Itoh Ichiro Yasuda Colonic TRPV4 overexpression is related to constipation severity BMC Gastroenterology Colon TRPV4 E. faecalis E. coli Chronic constipation |
| title | Colonic TRPV4 overexpression is related to constipation severity |
| title_full | Colonic TRPV4 overexpression is related to constipation severity |
| title_fullStr | Colonic TRPV4 overexpression is related to constipation severity |
| title_full_unstemmed | Colonic TRPV4 overexpression is related to constipation severity |
| title_short | Colonic TRPV4 overexpression is related to constipation severity |
| title_sort | colonic trpv4 overexpression is related to constipation severity |
| topic | Colon TRPV4 E. faecalis E. coli Chronic constipation |
| url | https://doi.org/10.1186/s12876-023-02647-0 |
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