Effects Of Mongolian Medicine Baoru On Intestinal Mucous Layer And TLR4/NFκB Signaling Pathway In Rats With Ulcerative Colitis

Objective:To investigate the effects of Mongolian medicine Baoru on intestinal mucous layer and TLR4/NFκB signaling pathway of TNBS/ ethanol-induced ulcerative colitis model rats。Methods:Seventy-two SPF-scored male SD rats were stochastically divided into six groups,including control group(Z),model group(M),Baoru low dose group(BD),Baoru medium dose group(BZ),Baoru high dose group(BG),and mesalazine group(MS),12 per group.After 1 week of adaptive rearing,rats in each group were all fat.Then without water for24 h,except for the blank control group,the remaining rats were molded with TNBS/ethanol solution according to the data,The same amount of distilled water in the control group.From the 4th day after modeling,each group of mice was administered enema according to the corresponding dose,once a day for 14 consecutive days.During the treatment process,the rats were observed to drink water and eat food,hair,mental state,Weight loss,poorly formed stools,blood in stools and DAI scores were performed.The rat colon was taken the day after the treatment,and the naked eye was observed to see whether the rat colon was involved in local congestion,ulceration,edema,etc.IHC method and RT-q PCR were used to see the protein expression and m RNA expression of MUC2 and TFF3 in the mucus layer in the colon.Western blot method and RT-q PCR method to observe the protein and m RNA expression of inflammatory signaling pathways TLR4 and NFκB.Result:1.General condition of ratsAfter one week of acclimatization,the weight of rats in each group increased steadily,there was no abnormality in diet and water,shiny hair,and normal feces.Compared with the normal group,TNBS ethanol enema rats had symptoms such as weight loss,reduced dietary water intake,gray hair,and r don’t like to move,DAI scores were significantly higher than before modelling,with a statistically significant difference(P < 0.01).After enema administration,DAI scores were relatively lower in all treatment groups except the model group,with a Obviously difference(P < 0.01).There was no significant difference between the Baoru high dose group and the mesalazine group(P > 0.05).2.Pathological changes in the colon of ratsThe colonic tissue structure of rats in the blank control group was basically normal,and no obvious pathological changes were observed.Pathological changes in the colon were evident in the model group,with the most severe damage;in the Baoru high-dose group,and mesalazine group were not obvious,and the damage was mild.3.Expression levels of MUC2 and TFF3 in the mucus layer of rat colonThe results of IHC and RT-q PCR showed that MUC2 and TFF3 protein expression and gene expression were significantly lower in the model group compared to the blank control group(P<0.01),while MUC2 and TFF3 protein expression and gene expression were significantly increased in the Baoru high-dose and mesalazine groups compared to the model group(P0.01 or P0.05),and there was no difference between the Baoru high-dose and mesalazine groups(P>0.05).Baoru low and medium dose groups between the model group‘s MUC2 protein expression,gene expression and TFF3 gene expression was no significant difference(P>0.05).4.Expression levels of TLR4 and NFκB in rat colon inflammationThe results of Western-blot and RT-q PCR showed that TLR4 and NF κ B protein expression and gene expression were significantly higher in the model group compared with the control group(P<0.01 or P<0.05),and TLR4 and NFκB protein expression 、gene expression were significantly lower in the Baoru high-dose group and mesalazine group compared with the model group after drug treatment(P<0.01 or P<0.05).There was no significant difference between the Baoru high dose group and the mesalazine group(P>0.05);Baoru low and medium dose groups ’s NFκB protein,gene expression and TLR4 gene expression compared with the model group were no significant(P>0.05).Conclusion:The general condition,disease activity index,and histopathological changes of TNBS/ ethanol-induced ulcerative colitis model rats were significantly improved,and its mechanism of action may be related to the maintenance of mucus layer structure and repair tissue damage by regulating factors such as MUC2 and TFF3 in the colonic mucus layer.On the other hand,it regulates the immune inflammatory response by inhibiting the release of factors related to TLR4,NF-k B and other factors related to important inflammatory signaling pathways.