| Purpose:By detecting the changes of STAT3,RORγt,TGF-β in colonic mucosa and IL-6,IL-10,IL-17 in serum of Ulcerative Colitis rats,the effect of electroacupuncture“Tianshu” and “Zusanli” on RORγt/STAT3 signaling pathway and immune imbalance were observed.To explore the mechanism of action of acupuncture and moxibustion,and provide certain experimental basis for acupuncture treatment of UC.Material and method:For SPF SD male rats were randomly divided into model group,blank group,non-meridian and non-acupoint group and electroacupuncture group by SPSS,with 10 rats in each group.Rats in model group,electroacupuncture group and non-meridian and non-acupoint group were fed 4% Dextran Sodium Sulfate solution,and the blank group was fed distilled water.From the fourth day,acupuncture was performed on“Tianshu” and “Zusanli” of rats in the electroacupuncture group.Acupuncture was performed at the midpoint of the line between “Tianshu” and the fourth nipple of rats in the non-meridian and non-acupoint group and at the lateral opening of “Zusanli” 0.5cm,with the waveform frequency of 2/15 Hz.The current intensity was 0.4-0.6m A,and the needle was kept for 15 minutes once a day for a total of 7 days.The body weight,fecal characteristics and blood in stool of rats in each group were recorded every day,and the disease activity index was calculated.After all the procedures were completed,the rats were treated by laparotomy,and then blood was collected from the abdominal aorta and colon tissue,and the degree of lesions in the colon mucosa was observed.The contents of IL-17,IL-10 and IL-6 in serum of rats were determined by ELISA;The content of STAT3,RORγt and TGF-βin colonic mucosa was determined by Western blot and the distribution of positive cells was observed by immunohistochemistry.Results:1.Effect of electroacupuncture early intervention on general state and pathology of UC rats(1)The general state of rats in the model group was worse than that in the blank group.Rats in the model group had dark and rough hair,thin stool or watery stool with blood color or even pus and blood stool,weight loss,and DAI index was higher than that in the blank group(P<0.05);The general state of the electroacupuncture group was better than that of the model group,and DAI index was lower than that of the model group(P<0.05);There was no significant difference between the general state and DAI index between the model group and the non-meridian and non-acupoint group(P>0.05).(2)The pathological comparison between the model group and the blank group showed that the colonic mucosa of the model group rats was discontinuous,accompanied by obvious bleeding,the number of glands decreased,hyperplasia of mucosa and submucosa,and a large number of inflammatory cell infiltration.After electroacupuncture treatment,the histomorphological characteristics of the colon of rats in the electroacupuncture group were significant improved,with no bleeding,the number of glands increased,the continuity of the mucosa was restored,and the number of inflammatory cells infiltrating in the submucosa and basal layer was reduced.There was no significant difference between the non-meridian and non-acupoint group and the model group.2.Effects of electroacupuncture on TGF-β,STAT3,RORγt,and IL-6,IL-17,IL-10 in UC rats.(1)The changes of IL-6,IL-17 and IL-10 in the serum of Ulcerative Colitis rats are:compared with the blank group,electroacupuncture group,model group,IL-6 without acupoints,IL-17 content increased,and the IL-10 content decreased(P<0.05);compared with the model group,the IL-6 content of the electroacupuncture group,non-acupoint group decreased(P<0.05),and the IL-10 content increased(P<0.05);the content of IL-6,IL-17 was higher than that of the non-acupuncture group and the electroacupuncture group(P<0.05).(2)The changes of STAT3,RORγt and TGF-βin the colonic mucosa of rats with Ulcerative Colitis were as follows:compared with the blank group,the contents of STAT3 and RORγt in the colonic mucosa of rats in the model group,the electroacupuncture group and the non-meridian and non-acupoint group were increased and the colonic mucosa was deepened,while the TGF-β content was decreased and the colonic mucosa was shallow(P<0.05).Compared with the model group,the contents of STAT3 and RORγt in the colon mucosa of rats in the electroacupuncture group and non-meridian and non-acupoint group were decreased,and the staining was shallow,while the content of TGF-β was increased,and the staining was deepened(P<0.05).Compared with the electroacupuncture group,STAT3 and ROR γ t contents increased and the pigmentation became darker,while TGF-β content decreased and the pigmentation became lighter in the non-meridian and non-acupoint group(P<0.05).Conclusion:1.Electroacupuncture at “Zusanli” and “Tianshu” can improve the body weight and DAI index score of intestinal histomphology inflammation in rats with ulcerative colitis.2.Electroacupuncture at “Zusanli” and “Tianshu” inhibited the expression of pro-inflammatory factors in serum and intestinal mucosa of rats with ulcerative colitis,and increased the expression of anti-inflammatory factors.The treatment of non-meridian and non-acupoint group is inferior to the electroacupuncture group.3.Electroacupuncture at “Zusanli” and “Tianshu” can not only activate the colonic mucosa of rats with ulcerative colitis,but also inhabit the expression of STAT3 and RORγt protein in the colonic mucosa,and make the immune balance axis return to a stable state.It has been preliminarily confirmed that acupuncture can regulated the immune balance mechanism. |
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