Effect Of "Tiao Zang Tong Luo" Electroacupuncture Therapy On Gut Microbiota In Db/db Mice

Objective:To investigate the effects of“Tiao Zang Tong Luo”electroacupuncture on type 2 diabetes mellitus by observing the mechanism of“Tiao Zang Tong Luo”electroacupuncture on body weight,blood glucose,gut microbiota structure and gut microbiota metabolites（acetic acid,propionic acid,butyric acid,valeric acid）in type 2diabetic db/db mice.Methods:22 db/db type 2 diabetic mice were randomly selected and divided into 11 for each group in the acupuncture group and the model group,and 9 db/m mice were blank controls,for a total of 3 groups.Before acupuncture treatment,1 week after acupuncture treatment and two weeks after,the tail blood glucose of each group was measured,the body weight of each group was weighed,and the feces of each group were taken and placed in-80°C refrigerator storage for inspection.The feces collected at different time periods were subjected to 16S ribosomal DNA identification,and the mouse fecal short-chain fatty acids,include butyric acid,acetic acid,valeric acid,and propionic acid,were detected by gas chromatography.The results were analyzed using statistical software SPSS 19.0.Results:1 General conditions of experimental animals and change of related indicators.1.1 General condition of experimental animalsSimilar to human diabetes,diabetic mice have the biological characteristics of polydipsia,polyphagia,polyuria.Blank controls:The mice are generally in good condition,with good coat color gloss,sensitive response,and normal food intake and drinking.Model control group:the gloss of the coat is poor,the response is sluggish,and the symptoms of polydipsia,polyphagia,and polyuria are apparent obviously.Acupuncture group:hair color gloss is not good,the reaction is slow,and food and water consumption and urine volume are obviously increased.Food intake:After 1 week and 2 weeks of intervention,compared with the blank group in the same period,the food intake of the acupuncture group and the model group increased significantly(^aaaP<0.001),while the food intake of the model group increased more significantly;Compared with the model group,the food intake of the acupuncture group was significantly lower than that of the model group（^bbbbb P<0.001）.Water intake:After 1 week and 2 weeks of intervention,compared with the blank group in the same period,the drinking volume of the acupuncture group and the model group increased significantly(^aaaP<0.001),while the increase of drinking water in the model group was more significant;compared with the model group,the increase in drinking water in the acupuncture group was significantly lower than that in the model group(^bbbP<0.001).1.2 The changes of body weight and blood glucose in each groupBody weight:Before the intervention,there was a significant difference in body weight between the acupuncture group and the model group and the blank group (^bbbP<0.001).There was no difference between the acupuncture group and the model group before intervention（P>0.05）.After two weeks of intervention,the acupuncture group had a slower weight gain compared with the model group,which indicates a significant difference between the two groups(^aaP<0.01).Blood glucose:Before the intervention,there was a difference in blood glucose between the acupuncture group and the model group and the blank group(^bbP<0.01).There was no difference between the acupuncture group and the model group before treatment（P>0.05）;After two weeks of intervention,the acupuncture group had a slower weight gain compared with the model group,which indicates a significant difference between two groups（^aP<0.05）.2 Changes of gut microbiota in each group of miceAfter two weeks of acupuncture treatment,in terms of Chao1 index,observed species index,acupuncture group is higher than blank controls,which is higher than model group.In terms of Simpson index and Shannon index,acupuncture group is higher than blank controls,which is higher than model group.In terms of Simpson index;After acupuncture intervention,at phylum level,relative abundance of Bacteroidetes and Fimicetes increased but relative abundance of Proteobacteria decreased,and Bacteroidetes/Fimicetes ratio decreased as well.At the genus level,the relative abundance of Lactobacillales,Lachnospiraceae and Prevotellaceae increased after acupuncture treatment,but the relative abundance of Escherichia and Betaproteobacteria decreased.3 Changes of short-chain fatty acids in gut microbiota metabolites of mice in each groupBefore the acupuncture intervention,the acetic acid content was reduced in the acupuncture group and the model group compared with the blank controls（cP<0.05、bP<0.05）.Before acupuncture intervention,there was no difference between the model group and the acupuncture group,which was statistically significant（P>0.05）;After acupuncture treatment,the acetic acid content was increased in the the acupuncture group compared with model group and significantly different（cP<0.05）;Compared with the blank controls,the propionic acid content of the acupuncture group and the model group were both increased,and the model group had more significant differences（aaP<0.01）.After treatment,there was a significant difference between the acupuncture group and the model group（bbP<0.05）;Before treatment,compared with the blank group,butyric acid in acupuncture group and model group decreased（aP<0.05）,and after treatment the content in acupuncture is significantly higher than model group（bP<0.01）.There was a significant difference between the acupuncture group before and after treatment（cP<0.05）;After treatment,the content of valeric acid shows decreased in the model group compared with the blank controls to reduce（aP<0.05）,but shows no difference compared with the acupuncture group.Conclusion:1.The db/db mice model of type 2 diabetes,similar with human syndrome of diabetes,were selected.Both body weight and blood glucose are elevated,so as food intake and drinking water.“Tiao Zang Tong Luo”electroacupuncture can inhibit the feeding and drinking of diabetic mice,and can improve the body weight and blood glucose of diabetic mice.2.“Tiao Zang Tong Luo”electroacupuncture increases the abundance and diversity of gut microbiota,reduces Bacteriodetes/Fimicetes ratio;Beneficial bacteria,such as Lactobacillales,had icreased and pathogenic bacteria,such as Escherichia,had been inhibited.The gut microbiota structure of the acupuncture group,compared with model group,was closer to the blank group.It explains that the acupuncture has a positive regulation effect on the gut microbiota of diabetic mice.3.“Tiao Zang Tong Luo”electroacupuncture can increase the content of acetic acid and butyric acid in the gut microbiota metabolites,reduce the content of propionic acid,and the valeric acid content does not change significantly.It indicates that acupuncture can regulate short-chain fatty acids and has an improved effect on diabetes.