| Purpose:By detecting the amount of DAF,FOXP3,TGF-? in the colonic mucosa of ulcerative colitis rats and the expression pattern,the changes of IL-6,IL-10,IL-17 content in serum of ulcerative colitis rats,are studies to check whether acupuncture at “Tianshu” and“Zusanli” could be interfered by FOXP3 / DAF signaling pathway and affected by the Treg /Th17 immune balance to play a role in the treatment of ulcerative colitis.To study the relationship between FOXP3 and DAF and to explore the relationship between acupunctures,the mechanism of action is investigated intending to lay the foundations for subsequent researches and to provide a new basis for clinical treatment of ulcerative colitis with acupunctures.Material and method:Forty SPF-grade SD rats were selected and randomly divided into 4 groups using SPSS software: blank group,model group,electroacupuncture group,non-acupoint group,with 10 rats in each group.Apart from the blank group,other groups used 4% dextran sodium sulfate in the drinking water for seven days in creating the rat model of ulcerative colitis.The treatment started on the fourth day after the model was developed.The electroacupuncture group selected rats for "Tianshu" and "Zusanli" electroacupuncture intervention.The non-acupoint and non-acupoint group selected "Tianshu" and the midpoint of the line connecting the fourth nipple on the same side."Zusanli" was opened by 0.5cm for electric acupuncture intervention for a total of seven days.During the intervention,the body weight,stool characteristics,and blood in the stool of each group of rats were recorded daily to calculate the disease activity index.After the treatment,all rats were anesthetized and treated with.Blood was taken from the abdominal aorta.The colonic mucosal tissue of each rat group was observed and the degree of colonic mucosal lesions in each group of rats was observed.The contents include IL-6,IL-10 and IL-17.The content and expression of DAF,FOXP3 and TGF-? in the intestinal mucosa of UC rats were measured by immunohistochemistry.The DAF in intestinal mucosa of UC rats was measured by western blotting and FOXP3.Results:1.Effects of pre-electroacupuncture on UC rat model with DSS(1)Compared with the blank group,the general condition of the rats in the model group was getting worse;rat fur became darker and duller than that in the normal group.Additionally,the feces were thinner or more watery than that in the normal group,and the bloody stools could be seen,and the body weight was significantly reduced.The disease activity index significantly increased(P <0.05);the general condition of the UC rats in the electroacupuncture group was better than that of the model group,and the weight increased compared with the model group,and the DAI score was significantly lower than that of the model group.The difference was statistically significant(P <0.05).There were no significant differences in general conditions,fecal traits,and blood in the stool between the acupoint group and the model group,and there was no significant difference between the DAI score and the model group(P> 0.05).(2)Colon pathological morphology results showed that compared with the blank group,the colonic mucosa in the model group was discontinuous with significant congestion;the number of glands decreased and the structure was destroyed;the number of shedding goblet cells increased,and the lower layer proliferated and there were a large number of inflammatory cells infiltration.After electroacupuncture intervention,the morphology of the colon tissue of the acupuncture group rats significantly improved;the number of glands increased;the continuity of the mucosal epithelium recovered;and the submucosal and basal layers of inflammatory cell infiltration was significantly less than that in the model group;there was no significant difference between the non-acupoint and acupoint group and the model group.2.The effects of electroacupuncture on DAF,FOXP3,TGF-? and IL-6,IL-10,IL-17 in serum of colonic mucosa of UC rats.(1)The serum ELISA results showed that compared with the blank group,IL-6 and IL-17 in the model group,electroacupuncture group,and nonacupoint group significantly increased(P <0.05),and all of IL-10 significantly decreased(P <0.05)).All IL-6 and IL-17 in the electroacupuncture group and non-acupoint group were lower than those in the model group(P <0.05),and all IL-10 were higher than the model group(P <0.05).Compared with the electroacupuncture group,L-6 and IL-17 were significantly increased(P <0.05),and IL-10 was significantly decreased(P <0.05).(2)The results of immunohistochemical staining of DAF,FOXP3,and TGF-? levels in the colonic mucosa showed that the positive cells of DAF,FOXP3 and TGF-? in the colonic mucosa of the model group UC rats were significantly lower than those of the blank group,and the coloring became lighter.Compared with the model group,DAF-positive cells in the colonic mucosa of the group were significantly reduced and the coloring became lighter.The DAF,FOXP3 and TGF-?-positive cells in the rat's colonic mucosa were significantly higher than the model group,and the coloring became darker.Although the Jingfei group was better than the model group,it was not as good as the electroacupuncture group.The positive cell ratio model group was significantly lower than the blank group,the result was statistically significant(P <0.05);the electroacupuncture group was higher than the model group but lower than the blank group,the result was statistically significant(P <0.05);The group was higher than the model group but lower than the electroacupuncture group,and the results were statistically significant(P <0.05).(3)Western blot analysis of DAF and FOXP3 levels in colonic mucosa showed that compared with the blank group,FOXP3 and DAF in the model group,electroacupuncture group,and non-acupoint group were significantly decreased,and the results were statistically significant(P <0.05).The electroacupuncture group and non-acupoint group were significantly higher than the model group,and the results were statistically significant(P <0.05);the electroacupuncture group was higher than the non-acupoint group,with statistically significant results(P <0.05).Conclusion:1.Electroacupuncture of "Tianshu" and "Zusanli" can effectively improve the weight,DAI score,morphological inflammation of intestinal tissue.Also it can increase the contents of TGF-?,FOXP3 and DAF in intestinal tissue,and increase IL-in serum in UC rats.The contents of IL10,reducing the levels of IL-6 and IL-17 in serum,although electroacupuncture at non-acupoints can increase the content of TGF-?,FOXP3 and DAF in the intestinal tissue, increase the content of IL-10 in serum,and reduce the contents of IL-6 and IL-17.The levels of IL-6 and IL-17 in the serum were not as good as those of the electroacupuncture "Tianshu" and "Zusanli".2.Electroacupuncture "Tianshu" and "Zusanli" have an effect on the FOXP3 / DAF signal pathway in the colonic mucosa,and preliminary proof of the mechanism of acupuncture to regulate the Treg / Th17 balance axis. |
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