A Study Of The Protective Effect Of Electro-Acupuncture On Gut Microflora In Rats With Ulcerative Colitis

Objectives:To study the abundence and diversity of the gut flora when treating ulcerative colitis by electro-acupuncture and to clearify whether electro-aucpuncture has the protective effect on gut microflora. Explore the advantages of acupuncture therapy on ulcerative colitis from the new perspective of the gut microflora to provide the basis of the mechanism of acupuncture in intestinal diseases,and provide new researching directions for the combination between acupuncture theory and the modern biology technique.Methods:39healthy male SD rats were divided into control group and model group randomly,.Ulcerative colitis model rats were induced by Dextran sulfate Sodium (DSS) with40g/L concentration. Then, the UC model group was randomly separated to UC model group and electro-acupuncture treatment group. The treatment group were acupunctured with "Tianshu""Zusanli" and "Shangjuxu". Each time the rats were treated for lOmin, once a day,5days as a treatment course.there were no treatment for the control and model group. Disease Activity Index (DAI) was used to evaluate the behaviors of the different group rats; benzidine assessment was used to detect the occult bleeding; tissue morphology was observed by light microscopy as well as ultrastructure was evaluated by transmission electron microscopy· observate the pathological and ultrastructure changes about the distal colon and ileocaecalis tissue with Light microscopic and electron microscopy. Stool of the rats were collect the rateds, stool, Denaturing Dradient Gel Electrophoresis was employed to anaalysis the composition and diversity of gut flora. The specific bands was obtained and sequenced strains results and data were analized by Quantity-one、Chromas、 SIMCA-P+、MGAE5and spss18.0softwares.Results:1. The observation results of therapeutic effect on electro-acupuncture:1). The DSS-induced UC model rats appeared poor general status even mucopurulent stool, DAI score significantly increased. After the treatment by electro-acupuncture, the mental activity and mucopurulent stool of the rats in electro-acupuncture group apparently approved, The DAI score decreased, there was statistical significant difference among groups(P<0.05).2). The observation results of UC model in the distal colon and ileocecus tissue by light microscopic,comparing with the blank group, show intestinal mucosal layer damage, ulcer, edema and congestion in submucosa, a large number of inflammatory cells infiltration, decrease of goblet cells, and lots of free necrotic tissue, while the electro-acupuncture group shows integrated structure of intestinal mucosal tissue, neat and tidy gland, no apparent edema and congestion, a few inflammatory cells infiltration, and hyperplasia of goblet cells.3). The observation results of UC model in the distal colon and ileocecus tissue by electron microscopy, comparing with the blank, group, appear shorter and looser microvilli, large vacuoles between epithelial cells, swelling mitochondria, large inflammatory cell infiltration. The eletro-acupuncture group relatively shows neat microvilli. normal mitochondria, evenly distributed nuclear chromatin, rich in rough endoplasmic reticulum.2. The protective effect of electro-acupuncture on gut microflora in rats with UC:1). Comparing with the blank group, the DGGE results show that the richness and diversity of intestinal flora in UC group was significantly reduced(P<0.05). While in the electro-acupuncture group, comparing with the UC group, the richness and diversity were apparently increased(P<0.05), and there was no statistical significant difference between this group and the blank group(P>0.05).2). The sequence results:the results of the samples after identifying mainly were Bacteroidetes, Firmicutes and Proteobacteria. The Bacteroidetes had been cultured bacteroides coprocola, and some had not cultured bacteroidetes. The Firmicutes had been cultured Lactobacillus sp., Lactobacillus johnsonii, Clostridium bifermentans. The Proteobacteria had been cultured Anaerobiospirillum succiniciproducens, Lawsonia intracellularis. The content of both Lactobacillus sp. and Lactobacillus johnsonii had been increased after electro-acupuncture treatment, which has statistical significant difference (P<0.05)comparing with the model group. The content of Clostridium bifermentans in model group significantly increased, which is higher than that of the blank group, while apparently decreased after electro-acupuncture treatment. Conclusions:1. There was a siginificant improvement in the Electro-acupuncture treatment group of the behaviors and the pathological characteristics of DSS-induced UC model rats.2. The abundance and diversity of intestinal bacteria in UC model rats was significantly lower than normal control group, electro-acupuncture can improve the abundance and diversity of the UC rat intestinal bacteria, indicating that the electric benign protective effect against intestinal bacterial diversity.3Electro-acupuncture can increases the content of Lactobacillus sp and Lactobacillus johnsonii, which are probiotics in the intestinal tract. Electro-acupuncture can also reduce Clostridium bifermentans and other pathogens of the gut. Our study indicate that electro-acupuncture play the role of protecting the gut microflora of ulcerative colitis.