| Aim: Diarrheal irritable bowel syndrome is one of the most common chronic diseases,and it has a significant effect on people’s lives.It is also a high-risk factor for type 2diabetes.Clinically,Tong Xie Yao Fang(TXYF)has a good therapeutic effect on diarrheal irritable bowel syndrome.In the present study,we constructed an inflammatory colonic mucosal cell model to study the effect of TXYF on antiinflammatory reactions.Meanwhile,the therapeutic effects and inflammation-related effects of TXYF were investigated by constructing liver-depressed spleen-deficient rats with D-IBS and T2 DM,and its mechanism of action was explored.The effects of TXYF on liver-depressed spleen-deficient rats with D-IBS and T2 DM were further analyzed by means of metabolomics.The aim of this study is to provide a basis for further demonstration of TXYF in D-IBS and T2 DM.Methods: Firstly,lipopolysaccharide(LPS)was selected to induce inflammation model of FHC colonic mucosal cells which were treated with different concentrations of TXYF-containing serum,and the effect of TXYF-containing serum on the activity of normal FHC cells and LPS-induced inflammatory FHC cells was detected by MTT method,followed by the detection of interleukin-6(IL-6)and interleukin-10(IL-10)expression using relevant kits.To study the effects of TXYF on liver-depressed spleendeficient rats with D-IBS and T2 DM,we first compared the relevant diarrheal markers,blood glucose values,and behavioral markers;then H&E staining was performed to detect the pathological changes in colonic tissues;then the changes of colonic tight junction protein claudin-1 were investigated by immunohistochemical method;The expression of tumor necrosis factor(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),and interleukin-10(IL-10)was then measured in colonic tissues by using relevant kits;and finally,the most relevant mechanistic pathways were enriched using network pharmacology and quantified by Western blot technique and relevant kits;High performance liquid chromatography(UHPLC)coupled with quadrupole electrostatic field orbitrap mass spectrometry(Q-Orbitrap-MS)was used to identify metabolites in rat serum and to investigate the differential metabolites and corresponding metabolic pathways between different groups.Results: TXYF increased the activity of FHC cells,restored the activity of FHC cells damaged by LPS and effectively counteracted the inflammatory damage of FHC cells induced by LPS,as shown by the decrease of IL-6 expression and the increase of IL-10 expression.In liver-depressed spleen-deficient rats with D-IBS and T2 DM,TXYF effectively alleviated the symptoms of diarrhea,improved emotions,and reduced blood glucose in the rats,and the results of H&E staining indicated that the colonic tissues of the treated rats were improved.The results of immunohistochemistry showed that TXYF may contribute to the repair of intestinal tight junction protein claudin-1.The changes in the expression of TNF-α,IL-1β,IL-6,and IL-10 consistently indicated that TXYF could effectively alleviate the inflammatory reactions in the colon of diseased rats.Network pharmacological analysis revealed that RAGE protein was most relevant to the mechanism of TXYF in treating liver-depressed spleen-deficient rats with D-IBS and T2 DM.The results of Western blot and the expression of MDA and SOD demonstrated that the action mechanism of TXYF was closely related to the RAGE oxidative stress pathway.Finally,the metabolomics analysis yielded 22 potential biomarkers and 7 metabolic pathways which were Purine metabolism,alpha-Linolenic acid metabolism,Fatty acid biosynthesis,Metabolic pathways,Starch and sucrose metabolism,Arachidonic acid metabolism,and Biosynthesis of unsaturated fatty acids.Conclusion: TXYF was able to increase the activity of FHC human colonic mucosal cells and counteract their inflammatory response.TXYF was effective in reducing blood glucose values and improving diarrhea and depression in liver-depressed spleendeficient rats with D-IBS and T2 DM.The results revealed that the RAGE pathway plays an important role in the treatment and TXYF modulates the expression of inflammatory factors through the RAGE oxidative stress pathway,thereby repairing the intestinal mucosal barrier.The results of the metabolic analysis showed that TXYF is also closely related to purine metabolism for the treatment of liver-depressed spleendeficient rats with D-IBS and T2 DM. |
PDF Full Text Request