"double Solid Through" Acupuncture On Diabetic Rats Neuroendocrine Immune Network Regulation Research

ObjectiveTo study the effect of "shuang gu yi tong ( tonifying congenital and acquired essence, eliminating pathogen)" acupuncture therapy on diabetes mellitus(DM) rats and the influence on the neuroendocrinoimmunity network of the DM rats, analyze its approaches to prevent the development of DM and its complications and explore its possible mechanism.MethodsDM models were made with Wistar rats by injecting streptozotocin into their lower left celiac cavity. Electrolyte lesion was used to injure PVN. 13 normal rats were divided into normal control group(group A).DM rats were randomly divided into four groups: model control group(group B), "yi tong" acupuncture group (group C), "shuang gu yi tong" acupuncture group (group D) and common acupuncture group (group E) , PVN lesion go up (group C'), "shuang gu yi tong" acupuncture and PVN lesion goup(group D'), sham lesion group(group E'). The treatment lasted for two courses, 6 days in each course and two days at interval. Blood sugar was tested with sugar equipment. T cell subtypes(CD4+/CD3 + ,CD8+/CD3+ ) were tested with flow cytometry. Serum insulin, IL-6 and leptin were measured with radioimmunoassy (RIA). Expressions of leptin receptor (LeptinR) and neuropeptide Y (NPY) in the hypothalamus of the rats were detected by immunohistochemistry SABC method. Expressions of LeptinR mRNA and NPYmRNA in the hypothalamus of the rats were detected by in situ hybridization. Image analyse system was used to measure the immune positive LeptinR, NPY and their mRNA.Results1 The therapeutic effect on the blood sugar of DM rats: In the treatment groups, the level of the blood sugar lowered after the treatment (P<0.05). The effect of group D and group E was better than that of group C. There was no significant difference between the effect of group D and group E (P>0.05). After the treatment there was no significant difference between the insulin level of group C, D, E and B. The level of insulin was lower than that of group A. The three treatment groups could reduce water and food intake and increase weight of the DM rats, among which the effect of group D was the best.2. Flow cytometry analyse indicated that: the ratio of CD4 + Tcell in model group was less than that of normal group, that of CD8 + Tcell was higher and the ratio of CD 4 + and C D 8 + T cell was lower than that o f thenormal group. After acupuncture treatment, the ratio of CD4+ T cell in group C, D and E increased, that of CD8+ T cell decreased and the ratio became higher. The regulative function of group D was the best among the groups.3. RIA results indicated that: serum IL-6 level of the DM rats was higher and leptin level was lower than that of the normal. There was no significant difference between the IL-6 levels of group C and group B. The effect of lowering IL-6 of group D was better than that of group E. The three treatment groups could increase leptin level of the DM rats, among which the effect of group D was the best.4. Immunohistochemistry and in situ hybridization results showed: Compared with the normal group, the expression of leptinR in hypothalamic paraventricular nucleus (PVN) and arcuate nucleus (ARC) and its mRNA in lateral hypothalamus (LH) were reduced significantly, the concentration of NPY in PVN and ARC and expression of its mRNA LH were increased significantly in STZ-induced diabetic rats. After acupuncture, the expression of leptinR and its mRNA were increased and the concentration of NPY and expression of its mRNA in hypothalamus were decreased obviously. The effect of group D was the best.5.Electrophysiologic research showed that: PVN lesion reduced food, water intake and leptin, improved weight of the DM rats. In group D', BG,food,water intake,weight and leptin were improved, while the effect was not good as that of group D. Sham lesion had no significant effect on the indice.Conclusion1. "shuang gu yi tong" acupuncture therapy has reliable effect of down-regulatin...