Study On The Regulation Mechanism Of Qingchangwen Chinese Recipe On Ulcerative Colitis Rats

Background:Withthe incidence and prevalence increased continuously,ulcerative colitis(UC)has become the hotspot and focus of current research,but the etiology and pathogenesis of UC is still unclear,soeffective means of treatment of UC islack.Traditional Chinese medicine has obvious advantages on the treatment of UC,especially in the stability of the disease,prevention of recurrence and other aspects,because it is based on syndrome differentiation,internal and external treatment,medication flexibility,andless toxic side effects.Therefore,the combination of traditional Chinese medicine and Western medicine will become the future direction of UC treatment.Qingchang Wenzhong Decoction(QCWZD)is a new and effective traditional Chinese medicine prescription formulated by Li Jun-xiang,a professor at the Beijing University of Chinese Medicine,can significantly improve the clinical symptoms of patients with UC,and repair endoscopic mucosa,then achieved remarkable curative effect in the clinical treatment of UC,but the specific mechanism is still not clear.Objective:The purpose of this experiment is to investigate the effect of QCWZD on DSS induced ulcerative colitis in rats,and with the MSP/RON signal pathway and IP10/CXCR3 axis as the starting point,to determine whether QCWZD treat ulcerative colitis by regulating MSP/RON pathway and IP10/CXCR3 axis.In addition,to clarify the effect target of QCWZD on ulcerative colitis,and provide experimental basis for further clinical application of QCWZD in the treatment of ulcerative colitisMethod:Experiment one:The rat model of ulcerative colitis was constructed by free drinking 4.5%DSS.After 7 days of intervention,the disease activity index(DAI)was evaluated,HE staining and histopathological score were performed,and the activity level of MPO was detected.Experiment two:The rat model of ulcerative colitis was constructed by free drinking 4.5%DSS.After 7 days of intervention,gene and protein expression of Occludin、ZO-1、Claudin-1、Claudin-2、Claudin-4were detected by Rt-PCR and Western Blot.Experiment three:The rat model of ulcerative colitis was constructed by free drinking 4.5%DSS.After 7 days of intervention,the expression levels ofcolonic inflammatory cytokines(IL-11,IL-1β,IL-6,IL-17,TNF-α,IFN-γ)and chemokineswere(MCP-1)detectedbyElisa.Experiment four:The rat model of ulcerative colitis was constructed by free drinking 4.5%DSS.After 7 days of intervention,serum MSP levels were detected by Elisa;WB was used to detect the expression of RON,AKT and p-AKT;and the protein distribution of RON and p-AKT was detected by IHC,respectively.Experiment five:The rat model of ulcerative colitis was constructed by free drinking 4.5%DSS.After 7 days of intervention,serum IP 10 level was detected by Elisa;Rt-PCR,WB and IHC were used to detect the gene and protein expression of IP 10,CXCR3,NF-kappa B,respectively.Result:Experiment one:When drinking a solution of 4.5%DSS,on the second day,the rats in the model group appeared positive fecal occult blood,and the naked eye can see the blood,stool soft start on the fourth day.With the passage of time,rats stool become heavier aggravating,the stool began to change into loose characters,body weight began to decline,the hair color gradually lose luster,the activity of the poor response is relatively slow,lazy.Blood in the stool,stool,weight loss,hair color and activity are the most serious on the seventh day.After QCWZD and mesalazine intragastric administration,the rats of the blood in the stool,traits,weight,hair color and activity were better than those in the model group.Compared with the normal group,DAI index,histological score and MPO levels of model groupincreased significantly compared with normal group(P<0.01),and after 7 days of QCWZD intervention,DAI index,histological score and MPO levels were significantly decreased(P<0.01,P<0.05,respectively).Experiment two:The gene and protein of occludin、ZO-1、claudin-1、claudin-4 in DSS-induced UC ratswere significantly lower than that in normal group,gene and protein expression of Claudin-2 was significantly higher,and after 7 days of QCWZD intervention,the gene and protein expressionof occludin、ZO-1、claudin-1、claudin-4increased significantly and Claudin-2expressiondecreasedsignificantly(p<0.01,p<0.05,respectively).Experiment three:The expression levels of colonic inflammatory cytokines(IL-1β,IL-1β,IL-6,IL-17,TNF-β,IFN-γ)and chemokines(MCP-1)in DSS-induced UC ratswere significantlyhigher than that in normal group,and after 7 days of QCWZD intervention,inflammatory cytokines andchemokinesexpression levelsdecreasedsignificantly(p<0.01,p<0.05,respectively).Experiment four:The level of serum MSP and the expression of colonic RON in DSS-induced UC ratswere significantly lower than those in control group,while the p-AKT/AKT level increased significantly(P<0.05,P<0.01,respectively).After 7 days of intervention,QCWZD significantly promote the expression of MSP,RON,and reduce the level of p-AKT/AKT(P<0.05,P<0.01,respectively).Experiment five:The level of serum serum IP 10,colonic IP 10,CXCR3 and NF-κBin DSS-induced UC ratswere significantly higher than those in control group significantly(P<0.05,P<0.01,respectively).After 7 days of intervention,QCWZD significantly reduce the level of serum IP10,colonic IP10,CXCR3 and NF-κB(P<0.05,P<0.01,respectively).Conclusion:1.QCWZD can reduce the DAI index and the down-regulatethe level of MPO.QCWZD has good therapeutic effect on DSS induced UC rats.2.The treatment of ulcerative colitis with QCWZD may regulate the expression of occludin,ZO-1,claudin-1,Claudin-2 in intestinal mucosal barrie,so as to enhance the intestinal mucosal barrier function and repair intestinal mucosal damage.3.QCWZD can downregulate the expression of inflammatory factors and chemokines,and then inhibit inflammatory reaction.4.QCWZD may enhance the function of MSP/RON pathway,promote its anti-inflammatory effect and repair intestinal mucosal barrier function,and finally achieve the purpose of treating UC.QCWZDmay be the activator of MSP/RON pathway.5.QCWZD ameliorates dextran sulphate sodium-induced ulcerative colitis in rats by downregulating the IP10/CXCR3 axis-mediated inflammatory response.QCWZD may be an inhibitor of the IP10/CXCR3 axis.6,QCWZD ameliorates DSS-induced ulcerative colitis in rats by enhancing the anti-inflammatory and intestinal mucosal repair function of MSP/RON pathway,meanwhile,downregulating the IP10/CXCR3 axis-mediated inflammatory response.