| ObjectivesObesity is a disorder of body energy metabolism caused by a variety of factors,resulting in fat cell hyperplasia or fat tissue content above the average level,which affects human health.Intestinal immune homeostasis disorders caused by intestinal microbiota and intestinal mucosal disorders caused by inflammatory activation are closely related to the development of Insulin Resistance.Changes in intestinal microbiota promote intestinal barrier dysfunction,activate inflammatory processes such as intestinal and peripheral adipose tissue,and promote Insulin Resistance(IR).This study intends to choose "specimen with hole" electric acupuncture and cupping benign bacteria after transplantation of intervention model of insulin resistance in obese Wistar rats,observe the lipid metabolism,insulin sensitivity index changes,the intestinal morphology,intestinal tight junction protein,the protein and m RNA expression of Occludin ZO-1,the structure of intestinal flora and abundance changes,thus the curative and fecal bacteria transplanted to insulin resistance and the regulation of intestinal barrier function,including for clinical use of prevention and treatment of insulin resistance in obesity and related metabolic diseases,to provide experimental basis,to explore the curative the potential values of benign bacteria after transplantation.MethodsSeventy Wistar male rats aged 5 weeks were divided into normal group(n=13)and model group(n=57)by random number table method,with normal and high fat feed,feed after 8 weeks,normal group of randomly selected three,building group randomly six,line is high insulin-glucose clamps,model assessment of insulin resistance,and to measure each rat body weight,anal nose long,calculate Lee’s index,assess obesity model,at the same time the evaluation standard is insulin resistance in obese rats building success.After successful modeling,rats in the modeling group were randomly divided into model group and electroacupuncture group,with 10 rats in each group.Each group was given corresponding intervention at the 10 th week of the experiment,and the normal group was fed with normal diet without any treatment.The model group was fed with high-fat diet without any treatment.The electroacupuncture group was fed with high-fat feed,and Zhongwan,Guanyuan,bilateral Fenglong and Zusanli acupoints were selected for electroacupuncture intervention.Hans LH202 H electroacupuncture instrument was used,the intensity was 1m A,the frequency was 2Hz,continuous wave,electroacupuncture intervention 10 minutes/time,3 times/week,a total of 8 weeks of intervention.In addition,another 30 5-week-old Wistar male rats were purchased at the 9th week of the experiment.After adaptive feeding for a week,they were fed with high-fat diet for 8 weeks.After 8 weeks,6 rats were randomly selected for hyperinsulin-positive glucose clamping operation.After successful modeling,rats were randomly divided into model control group and faecal bacteria transplantation group,with 10 rats in each group.Model control group was fed with high fat diet and saline enema.The fecal bacteria transplantation group was fed with high-fat diet,and the fresh feces of the rats after electroacupuncture intervention were processed into fecal bacteria solution and then enema was given,once a week,for 3 weeks.The body weight,anal nose length and Lee’s index of all rats were measured before,2,4,6 and 8 weeks after intervention.IPGTT and IPITT were detected at the 6th week of the experiment.After the intervention,samples were collected.Before death,fresh feces of rats were collected for acer detection.Serum samples were collected from apical blood,and serum insulin(INS)and endotoxin(LPS)levels were detected by ELISA.After death,colon tissues of rats were stored in a refrigerator at-80℃.Colon tissues of rats in each group were sected and stained with HE,and observed under light microscope.Western Blot was used to detect the protein expression levels of ZO-1 and Occludin in the colon of rats,and PCR was used to detect the gene expression levels of ZO-1 and Occludin.Results1.Effects of high fat diet on body weight,Lee’s index and GIR of rats: after 8 weeks of feeding,the body weight and Lee’s index of rats fed with high fat diet were higher than those fed with ordinary diet(P < 0.01);The mean GIR of rats fed with normal diet was lower than that of rats fed with normal diet(P <0.01).2.Effects of electroacupuncture and bacterial community transplantation after electroacupuncture on insulin sensitivity of rats: After electroacupuncture treatment for 8 weeks and faecal bacteria transplantation after electroacupuncture for 3 weeks,both of them can reduce body weight,Lee’s index,serum LPS and INS content of obese rats with IR,and improve insulin sensitivity of rats.3.Electric acupuncture and cupping flora after transplantation of colonic mucosa of rats and flora structure: the effect of electric acupuncture and cupping flora after transplantation can improve the colonic mucosa structure,morphology and flora improve bacteroidetes abundance,reduce the thick wall bacteria abundance,and according to beta diversity analysis,model group and normal group in the rat intestinal flora structure difference,but after acusector and fecal bacteria transplantation treatment group the structure of rats and normal rats more similar.4.Influence of electroacupuncture and bacterial colony transplantation after electroacupuncture on ZO-1 and Occludin in colon of rats: Electroacupuncture and bacterial colony transplantation after electroacupuncture can effectively up-regulate the expression levels of tight junction proteins ZO-1,Occludin and their m RNA in colon of insulin resistant obese rats.Conclusions1.Continuous feeding of a high-fat diet can successfully induce the establishment of an insulin resistant obese rat model.Long-term high-fat diet can cause changes in intestinal microbiota structure and affect intestinal barrier function.2.Electroacupuncture and bacterial community transplantation after electroacupuncture can reduce the weight and degree of obesity in IR obese rats,improve insulin sensitivity,reduce the release of endotoxin LPS,and improve insulin resistance in rats.3.Electric acupuncture and cupping can protect bacteria after colonic mucosa form,improve the intestinal microbiota abundance and diversity,and improves the beneficial bacteria in the gut microbiota abundance and abundance,and reduce pathogenic bacteria through the beta diversity analysis,the results show that after the local area and cupping flora transplantation in the rat intestinal microbiota structure similar to the normal group,electric acupuncture can adjust the intestinal barrier function and intestinal microbiota structure to improve IR.4.Both electroacupuncture and microflora transplantation after electroacupuncture can up-regulate the expression of colon tight junction proteins ZO-1 and Occludin,and improve the function of intestinal mechanical barrier. |
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