Résumé
Objective To investigate the effect of Yiqi Huoxue Huatan Decoction(YHHD) on gene expression of vascular endothelial growth factor(VEGF), intercellular adhesion molecule 1(ICAM-1) in the colonic mucosa of ulcerative colitis rats. Methods Sixty rats were divided into six groups, namely normal control group, model control group, low-, middle- and high-dose Y HHD group(gastric gavage of YHHD in the dosage of 13, 26, 52 g·kg-1·d-1 respectively) , and Mesalazine(0.5 g·kg-1·d-1 ) group. T he rat model of ulcerative colitis was established by enema with 2, 4, 6-trinitrobenzene sulfonic acid(TNBS)mixture(2.5%TNBS and 50% ethanol). The changes in rat body weight were monitored. After treatment, pathological features of the colonic mucosa were observed under light microscope, and VEGF and ICAM-1 gene expression levels in the colonic mucosa were measured by reverse t ranscripti on polymerase chai n reaction(RT-PCR). Results Body weight was lighter, and relative gene expression levels of VEGF and ICAM-1 in the model group were significantly higher than those in the normal control group, and pathological features of the colonic mucosa was obvious, the differences being significant between the two groups(P<0.01). Compared with the model group, body weight was increased , pathological features of the colonic mucosa were relieved, and relative gene expression levels of VEGF and ICAM-1 were significantly decreased in YHHD groups(P<0.01), and high-dose YHHD showed the strongest effect. Conclusion YHHD can down-regulate the gene expression of VEGF and ICAM-1, which may be one of its therapeutic mechanisms for ulcerative colitis.
Résumé
Objective To explore the biological mechanism of the Lingering Toxins Theory in ulcerative colitis (UC) by clinical trials. Methods Forty patients with UC at mild, moderate active stage and at remission stage were enrolled into the experiment. Twenty cases of UC patients at active stage were divided into Chinese medicine intervention group and western medicine treatment group, 10 in each group. Twenty cases of UC patients at remission stage were also evenly divided into Chinese medicine intervention group and western medicine treatment group. All patients were given conventional western medicine treatment, and additionally, Chinese medicine intervention group were given oral use of Chinese medicine decoction, the treatment course covering 8 weeks. Before and after treatment, plasma matrix metalloproteinase-1 (MMP -1), tissue inhibitor ofmetalloproteinase-1 (TIMP-1) and prostaglandin E2 (PGE2) expression levels were observed by enzyme-linked immunosorbent assay (ELISA) . Results Plasma MMP-1,TIMP-1, and PGE2 expression levels in UC patients were positively correlated with disease severity (P0.05) . After treatment, the expression levels of MMP-1 and TIMP-1 in remission-stage patients of Chinese Medicine intervention group were decreased (P0.05) . Conclusion There is a relation between plasma MMP-1, TIMP-1 expression levels and the “lingering toxins” in the field of traditional Chinese medicine. The biological mechanism of toxins-removing therapy in treating UC may be associated with the decrease of the plasma MMP-1 and TIMP-1 expression levels.
Résumé
Objective To study the preventive effect and mechanism of aesculin on intestinal mucosa in rats with experimental ulcerative colitis (UC) . Methods Forty specific-pathogen free SD rats were randomly divided into normal group, model group, salicylazosulfapyridine (SASP, 600 mg/kg) group and aesculin (EH, 100 mg/kg) group, 10 in each group. Rats in model group, SASP group and EH group were given enema with trinitrobenzene sulfonic acid ( TNBS, 100 mg/kg) for the establishment of UC model. The rats in SASP group and EH group were given gastric gavage of SASP and aesculin respectively. At the end of experiment, the serum levels of tumor necrosis factor alpha ( TNF-α) and interleukin 10 ( IL-10) were detected by enzyme-linked immunosorbent assay (ELISA) . The general state, histological features of intestinal mucosa and serum TNF-αand IL-10 levels of rats in each group were compared. Results Aesculin significantly improved the general state and relieved the inflammation of the colonic mucosa of UC rats. The disease activity index ( DAI) scores and tissue damage index (TDI) scores in the model group were significantly higher than those in the normal group ( P<0.01) . The DAI scores and TDI scores in the medication groups were significantly lower than those in the model group (P<0.01) . The serum TNF-αlevel was significantly higher and IL-10 level was significantly lower in the model group than the normal group ( P<0.01) . After treatment, TNF-α was decreased and IL-10 was increased in SASP group and EH group as compared with the model group (P<0.01) . Conclusion Aesculin has certain therapeutic effect on TNBS-induced UC in rats through significantly relieving the symptoms of UC rats. The mechanism may be related with the inhibition of TNF-α secretion and the increase of IL-10 expression, and then improving the disorder of intestinal immune function.
Résumé
Objective To investigate the cytokines levels in peripheral blood of ulcerative colitis (UC) patients at active stage, and to observe the effect of baicalin at various concentrations on the cytokines. Methods Quantitative polymerase chain reaction (Q-PCR) assay was used for the detection of interleukin 4 receptor (IL4R), IL6R, IL23R and RAR-related orphan receptor C (RORC) expression in single peripheral mononuclear cell of UC patients. Enzyme-linked immunosorbent assay (ELISA) was used to examine the serum levels of interferon gamma (IFN-γ), IL-4, IL-5, IL-6, IL-10, and transforming growth factor beta (TGF-β) levels in UC patients, and was applied for in-vitro detection of these indicators after baicalin intervention. Results Q-PCR results showed that IL4R, IL6R, IL23R, RORC gene expression levels in UC patients were increased to various degrees as compared to diarrhea-predominant irritable bowel syndrome (IBS-D) patients and the healthy volunteers, and then the levels were decreased to various degrees after intervention by baicalin at different concentrations, in particular at 20 and 40μmol/L. The results of ELISA showed that serum levels of cytokines of IFN-γ, IL-5, IL-6 in UC patients were increased to various degrees while IL-4, IL-10 and TGF-β1 was decreased as compared to IBS-D patients and the healthy volunteers. Baicalin at the concentrations of 20 and 40 μmol/L had an effect on decreasing IFN-γ, IL-5, IL-6 and on increasing IL-4 and IL-10. Conclusion Higher concentrations of baicalin show obvious effect on inhibiting RORC and IL23R expression, on decreasing IFN-γ, IL-5, IL-6 levels, and on increasing IL-4, IL-10 and TGF-β1 levels, which indicated that the therapeutic mechanism of baicalin in relieving ulcerative colonic inflammation is related with the regulation of immune function.
Résumé
【Objective】To observe the effect of Kuijieling Decoction(KD) on NF-?B p65 protein expression in colonic mucosa and serum tumor necrosis factor ?(TNF-?) content of ulcerative colitis rats.【Methods】Sixty SD rats were randomized into 6 groups: normal group,model group,salicylazosulfapyridine(SASP,0.5g?kg-1?d-1) group and three KD(18.3,9.2 and 4.6 g?kg-1?d-1 respectively) groups.Except the normal group,the rats received enteroclysis of trinitro-benzene-sulfonic acid(TNBS) to induce ulcerative colitis models.Serum content of TNF-? was detected by enzyme-linked immunosorbent assay(ELISA).Then the rats were executed to make tissue section.The tissue sections were stained by immunohistochemistry and the cells with positive expression of NF-?B p65 protein were counted under microscope.【Results】The serum level of TNF-? and the cell number of positive expression of NF-?B p65 protein in high-and moderate-dose KD groups were lower than those in the model group(P