RESUMEN
OBJECTIVE: To observe the effects of moxibustion at "Tianshu"ï¼ST25ï¼ and "Shangjuxu"ï¼ST37ï¼ on the colonic metabolites and inflammatory factors in rats with Crohn's diseaseï¼CDï¼, so as to explore the mechanisms of moxibustion in protecting colon of CD rats based on metabolomics. METHODS: Twelve rats were first randomly selected from 36 male SD rats as a normal groupï¼NGï¼. The CD model was induced by 2, 4, 6 trinitrobenzene sulfonic acidï¼TNBSï¼ enema on the rest 24 rats. After successful modeling, rats were randomly divided into modelï¼TNBSï¼ and moxibustionï¼TNBS+MOXï¼ groupsï¼n=10 rats/groupï¼. Moxibustion was applied at bilateral ST25 and ST37 for 30 min, once daily for 7 consecutive days in the TNBS+MOX group, while rats in the NG and TNBS groups did not receive any interventions. Body weight of rats was recorded and disease activity indexï¼DAIï¼ was assessed during the experiment. After interventions, HE staining was performed to observe pathological damage of colon. Serum levels of inflammatory factors were measured by ELISA. NMR hydrogen spectroscopy was used to detect colonic metabolites of each group, and orthogonal partial least squares discriminant analysisï¼OPLS-DAï¼ was used to screen differential colonic metabolites between groups, followed by pathway analysis using MetaboAnalyst 5.0 platform. RESULTS: After modeling, compared with the NG group, the body weight of the rats in the TNBS group was significantly decreasedï¼P<0.05ï¼, the DAI score was increased ï¼P<0.05ï¼, the colon had obvious inflammatory damage and the pathological injury index was increasedï¼P<0.05ï¼, and levels of serum tumor necrosis factor-αï¼TNF-αï¼, interleukinï¼ILï¼-1ß and interferon-γï¼IFN-Î³ï¼ were significantly increasedï¼P<0.05ï¼. After moxibustion intervention, compared with the TNBS group, the body weight was significantly increasedï¼P<0.05ï¼, while the levels of serum TNF-α, IL-1ß, IFN-γ, and DAI score of the rats in the TNBS+MOX group were significantly decreasedï¼P<0.05ï¼, with alleviated colonic inflammatory injury detected by HE staining. Compared with the NG group, the relative expressions of colonic hypoxanthine, betaine, creatine, inositol, taurine, uracil, and methanol of the TNBS group were decreasedï¼P<0.05ï¼, while the relative expressions of histidine, leucine, proline, lysine, isoleucine, phenylalanine, tyrosine, propionic acid, and valine were increasedï¼P<0.05ï¼ in the TNBS group, among which, relative expressions of hypoxanthine, leucine, lysine, isoleucine, betaine, tyrosine, and taurine were reversed in the TNBS+MOX group relevant to the TNBS group, mainly involving phenylalanine, tyrosine and tryptophan biosynthesis, and taurine and subtaurine metabolism pathway. CONCLUSION: The mechanism of moxibustion at ST25 and ST37 for CD may be related to improving colon metabolic disorder state by regulating multiple metabolic metabolites and metabolic pathways, and reducing the level of inflammatory factors, so as to maintain intestinal immune homeostasis.
