The Influence And Mechanism Of QingChangHuaShi Decoction Treating Model Rats Of Ulcerative Colitis

Objective The purpose of the research is to supply the objective evidences for clinical prescription and to pave the way for developing the new Chinese herbal drugs through exploring the mechanism of QingChangHuaShi decoction treating Ulcerative colitis. The observing targets include the treatment effectiveness on TNBS-induced model rats colitis, proinflammatory cytokines, intercellular adhesion molecule-1, dendritic cells tight junction protein, nuclear factor-KB of colonic mucous membrane and dendritic cells of mesenteric lymph nodes.Methods Forty male depuratory Wistar rats were randomly divided into four groups, and each group has ten rats.The weight of one rat is about200±20gram,and the age is about six to seven weeks. The blank group ten rats were raised commonly,eat and drink freedomly; The other thirty rats were divided into model group、QingChangHuaShi decoction group and Mesalazine group after successfully building the model induced by TNBS. The model group rats were gave equivalent volume physiological saline though intragastric administrations at one time everyday. The dosage of the QingChangHuaShi decoction and Mesalazine was12.8g dried medicinal herb·kg-1·d and0.67g·kg-1·d respectively. The administration methods were the same as the blank group. The administration time was ten days and the drugs were gave from the second day successively after building the models. Fasting for24h form the last administration, and then all of the rats were killed. The fresh colonic tissues and the MLNs were resected for detection. The general conditions and the colonic pathology morphology were observed. The proinflammatory cytokines including IL-6、IL-β、TNF-α were measured by the method of ELISA. The numbers of the DCs in the colonic mucous membrane and mesenteric lymph nodes were measured by the method of IHC. The ratio of the DCs in the MLNs and the expression of superficial molecule CD86and MHC-Ⅱwere checked out by the flow cytometer.ICAM-1and s ICAM-1were checked out by IHC and ELISA respectively. And the expression levels of the occludin mRNA、 claudin-1mRNA、NF-κB p65mRNA were measured by the method of the real-time PCR, and the occludin、claudin-1、NF-κB p65proteins by IHC. The NF-κB p65protein was also checked out by WB.At last,the correlation between the NF-κB p65and the inflammatory accumulated points、IL-1β, TNF-α, IL-6, Claudin-1、occludin, DCs and ICAM-1were evaluated by the correlation analysis.Results After modeling, the pathology inflammatory accumulated points of the colonic membrane and the expression level of proinflammatory cytokines including IL-6、IL-1β、 TNF-a,ICAM-1、sICAM-1, NF-κB p65mRNA and NF-κB p65protein increase significantly (P<0.01). The numbers of the DCs in the colonic mucous membrane and mesenteric lymph nodes and the expression of the superficial molecule MHC-Ⅱ of the DCs in the mesenteric lymph nodes increase significantly (P<0.01).And the expression levels of the occludin mRNA, claudin-1mRNA, occludin、claudin-1proteins decrease (P<0.05).After treatment, the pathology inflammatory accumulated points of the colonic membrane and the expression level of proinflammatory cytokines including IL-6、IL-1β、TNF-α,ICAM-1、sICAM-1, NF-κB p65mRNA and NF-κB p65protein decrease significantly (P<0.01). The numbers of the DCs in the colonic mucous membrane and mesenteric lymph nodes and the expression of the superficial molecule MHC-Ⅱ of the DCs in the mesenteric lymph nodes decrease significantly (P<0.01) And the expression levels of the occludin mRNA、claudin-1mRNA, occludin、claudin-1proteins increase (P<0.05). The expression of NF-κB p65intimately correlates with inflammatory accumulated points, IL-1β、TNF-α、IL-6, Claudin-1、occludin, DCs, ICAM-1(P<0.01).The ratio of the DCs in the MLNs and the expression of superficial molecule CD86have no statistical difference (P>0.05) between pretherapy and prior treatment among four groups,and the influences on all of the indices have no statistical difference (P>0.05) between QingChangHuaShi decoction and Mesalazine.Conclusion QingChangHuaShi decoction can improve general conditions and colonic inflammation of the TNBS-induced model rats. The concrete mechanism are as follows: suppressing the secretion level of proinflammatory cytokines such as IL-6、IL-1β、TNF-α, decreasing the numbers of the colonic membrane and the mesenteric lymph nodes, suppressing the activating of the DCs, reducing the expression level of ICAM-1,and increasing the expression of TJ proteins, thus to retrieve the function of intestine barrier, possibly through suppressing the expression of the nuclear factor-KB.