Zhongguo zhen jiu = Chinese acupuncture & moxibustion · 2025
[Mechanism of moxibustion on intestinal fibrosis in Crohn's disease based on Trp/Kyn/AhR pathways in mice].
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Gao Qianqian, Feng Qianru, Peng Yang, Qi Qin, Wu Huangan, Liu Huirong, Zhou Cili
Аннотация
OBJECTIVE: To observe the effect of moxibustion on tryptophan (Trp)/kynurenine (Kyn)/aryl hydrocarbon receptor (AhR) pathway in the colon of mice with intestinal fibrosis of Crohn's disease (CD), and explore the mechanism of moxibustion on CD intestinal inflammation and intestinal fibrosis.
METHODS: Among 28 male SPF C57BL/6 mice, 8 mice were randomly selected as the normal group, and CD intestinal fibrosis models were prepared by enema with 5% trinitro-benzene sulfonic acid (TNBS) in the remaining mice as the modeling group. Eighteen successfully modeled mice were randomly divided into a model group, a western medication group, and a moxibustion group, 6 mice in each one. In the moxibustion group, mild moxibustion was operated at "Qihai" (CV6) and bilateral "Tianshu" (ST25), once daily, 10 min each time and for consecutive 7 days. In the western medication group, the gavage with mesalazine enteric-coated tablet suspension, 0.5 mL was administered, once a day and for consecutive 7 days. The general condition, score of disease activity index (DAI), colon length, and colonic macroscopic score were observed in each group. HE and Masson staining were used to examine the morphology of the colon and intestinal fibrosis in mice, and the collagen volume fraction (CVF) was calculated. ELISA was adopted to detect the contents of interleukin (IL)-12, IL-23, Trp and Kyn in colon tissue. Using immunofluorescence assay, the positive expression of Collagen Ⅰ, Collagen Ⅲ and indolamine 2,3-dioxygenase (IDO) was observed. Using Western blot method, the protein expression of Collagen Ⅰ, Collagen Ⅲ, IDO and AhR in colon was detected. Using real-time quantitative PCR, the mRNA expression of IDO and AhR in colon was detected.
RESULTS: Compared with the normal group, in the model group, the body mass was reduced and the colon length shortened (P<0.01), DAI score, the gross macroscopic score and histological score of colon increased (P<0.01), CVF was higher (P<0.01), the contents of IL-12, IL-23, Trp and Kyn and the ratio of Kyn to Trp were elevated (P<0.01), the protein expression of Collagen Ⅰ and Collagen Ⅲ increased (P<0.01), and the protein and mRNA expression of IDO and AhR were higher (P<0.01). When compared with the model group, in the moxibustion group and the western medication group, the colon length was increased (P<0.01), DAI score, the gross macroscopic score and histological score of colon were reduced (P<0.05, P<0.01), CVF was lower (P<0.01), the contents of IL-12, IL-23, Trp and Kyn and the ratio of Kyn to Trp were reduced (P<0.01, P<0.05), the protein expression of Collagen Ⅰ and Collagen Ⅲ decreased (P<0.05), the protein and mRNA expression of IDO and AhR was declined (P<0.01, P<0.05). In comparison with the moxibustion group, the contents of Trp and Kyn in the colon were lower in the western medication group (P<0.01).
CONCLUSION: Moxibustion can attenuate intestinal inflammation and fibrosis induced by TNBS in CD mice, and its mechanism may be related to regulating Trp/Kyn/AhR pathway.
METHODS: Among 28 male SPF C57BL/6 mice, 8 mice were randomly selected as the normal group, and CD intestinal fibrosis models were prepared by enema with 5% trinitro-benzene sulfonic acid (TNBS) in the remaining mice as the modeling group. Eighteen successfully modeled mice were randomly divided into a model group, a western medication group, and a moxibustion group, 6 mice in each one. In the moxibustion group, mild moxibustion was operated at "Qihai" (CV6) and bilateral "Tianshu" (ST25), once daily, 10 min each time and for consecutive 7 days. In the western medication group, the gavage with mesalazine enteric-coated tablet suspension, 0.5 mL was administered, once a day and for consecutive 7 days. The general condition, score of disease activity index (DAI), colon length, and colonic macroscopic score were observed in each group. HE and Masson staining were used to examine the morphology of the colon and intestinal fibrosis in mice, and the collagen volume fraction (CVF) was calculated. ELISA was adopted to detect the contents of interleukin (IL)-12, IL-23, Trp and Kyn in colon tissue. Using immunofluorescence assay, the positive expression of Collagen Ⅰ, Collagen Ⅲ and indolamine 2,3-dioxygenase (IDO) was observed. Using Western blot method, the protein expression of Collagen Ⅰ, Collagen Ⅲ, IDO and AhR in colon was detected. Using real-time quantitative PCR, the mRNA expression of IDO and AhR in colon was detected.
RESULTS: Compared with the normal group, in the model group, the body mass was reduced and the colon length shortened (P<0.01), DAI score, the gross macroscopic score and histological score of colon increased (P<0.01), CVF was higher (P<0.01), the contents of IL-12, IL-23, Trp and Kyn and the ratio of Kyn to Trp were elevated (P<0.01), the protein expression of Collagen Ⅰ and Collagen Ⅲ increased (P<0.01), and the protein and mRNA expression of IDO and AhR were higher (P<0.01). When compared with the model group, in the moxibustion group and the western medication group, the colon length was increased (P<0.01), DAI score, the gross macroscopic score and histological score of colon were reduced (P<0.05, P<0.01), CVF was lower (P<0.01), the contents of IL-12, IL-23, Trp and Kyn and the ratio of Kyn to Trp were reduced (P<0.01, P<0.05), the protein expression of Collagen Ⅰ and Collagen Ⅲ decreased (P<0.05), the protein and mRNA expression of IDO and AhR was declined (P<0.01, P<0.05). In comparison with the moxibustion group, the contents of Trp and Kyn in the colon were lower in the western medication group (P<0.01).
CONCLUSION: Moxibustion can attenuate intestinal inflammation and fibrosis induced by TNBS in CD mice, and its mechanism may be related to regulating Trp/Kyn/AhR pathway.