| [Significance]Diabetes is a metabolic disease of the endocrine system caused by a combination of genetic and environmental factors.Type 2 diabetes accounts for more than 90%of the affected population.With economic development,improved living standards and changes in lifestyle,the prevalence of diabetes among obese people has increased significantly.Insulin resistance,as an important mechanism for the onset of type 2 diabetes,is manifested by the decreased sensitivity of peripheral tissues and organs to insulin.Skeletal muscle is the most important peripheral target organ of insulin.It is an important tissue for insulin-mediated uptake and utilization of glucose.80%of glucose in the blood circulation is taken up and metabolized by skeletal muscle.Therefore,it is particularly critical to study the mechanism of insulin resistance in skeletal muscle.Since the current hypoglycemic drugs all have certain side effects and adverse reactions,acupuncture has been proven by a large number of papers to effectively interfere with type 2 diabetes.Therefore,exploring the mechanism of electro-acupuncture to lower blood sugar and improve skeletal muscle insulin resistance is of great significance for the treatment of obese type 2 diabetes.[Objective]1.To observe the effects of electro-acupuncture on blood glucose,body weight,leptin,insulin,C peptide,blood lipids and other indicators in obese rats with type 2 diabetes,so as to explore the possible mechanism of electro-acupuncture in the treatment of type 2 diabetes.The effect of electro-acupuncture on the blood glucose regulation of type 2 diabetic rats will be verified,and whether acupuncture can improve the insulin resistance of type 2 diabetic patients will be tested.2.The purpose of this study is to explore whether electro-acupuncture can adjust the balance between skeletal muscle IRS-1 tyrosine and serine/threonine phosphorylation,promote downstream phosphatidyl inositol 3-kinase/protein kinase B(PI3K/Akt)pathway conduction,and up-regulate skeletal muscle glucose transporter 4(GLUT4)membrane shift to reduce blood sugar and normalize insulin resistance.To observe the effect of electricity on the level of IRS-1 phosphorylation in skeletal muscle of type 2 diabetic rats.The effects of electro-acupuncture on the regulation of IRS-1 tyrosine phosphorylation and serine/threonine phosphorylation in skeletal muscle cells were evaluated by Western blot.At the same time,detect the skeletal muscle PI3K protein level.Based on this,the GLUT4,a terminal indicator of the skeletal muscle insulin signaling pathway,is examined to investigate the possible mechanism of electro-acupuncture for type 2 diabetes.[Method]Seven 2 month old SPF male ZL(Zucker Lean)rats(fa/+)and 14 same month old SPF male ZDF(Zucker Diabetic Fatty)rats(fa/fa)were selected as experimental subjects.One week after the adaptive breeding,all rats were fed with high-sugar and high-fat induction feed for 4 weeks,and the animals were free to drink water.After 4 weeks of feeding rats with high-sugar and high-fat diets,the fasting blood glucose and OGTT2h blood glucose are more than 11.1 mmol/L as the model standard.Fourteen ZDF rats after modeling were randomly divided into a model group and an electro-acupuncture group,with 7 rats in each group;another 7 ZL(Zucker Lean)rats were set up as a control group.The control group and model group were not intervened with treatment.The electro-acupuncture group acupunctured bilateral Zusanli(ST36),Sanyinjiao(SP6),Weiwanxiashu(EX-B3),and Pishu(BL20)with a direct penetration depth of 4-6mm,and connected with an electro-acupuncture instrument(one end of the electric acupuncture is connected to Weiwanxiashu(EX-B3),and one end was connected to Zusanli(ST36)on the same side to form a loop).Continuous wave,frequency of 15Hz,current output intensity 2mA,it is advisable for rat muscles to shake slightly without screaming,leaving needles for 25min,6 times a week(Monday to Saturday),continuous intervention for 4 weeks.Fasting blood glucose(FBG)and body weight are measured before and after experiment modeling,and at the end of the fourth week of intervention.Serum is taken 4 weeks later,and triglycerides(TG),total cholesterol(TC),high-density lipoprotein(HDL-C),low-density lipoprotein(LDL-C),and very low-density lipoprotein(VLDL-C)are detected by colorimetry;observe serum leptin(LEP),insulin(INS),C peptide(CP)levels.Take part of the skeletal muscle tissue of the rats under the ice bath condition,and measure the levels of IRS-1 tyrosine and serine/threonine phosphorylation by Western Blot method;observe the skeletal muscle PI3K and whole cell GLUT4 levels.[Result]Experiment 1 shows:After 4 weeks of intervention,the fasting blood glucose of the electro-acupuncture group is lower than that of the model group,and the difference is statistically significant(P<0.05).It can be seen within the group that the fasting blood glucose of the electro-acupuncture group and the model group is higher than that before the modeling after the modeling and the intervention,the difference is statistically significant(P<0.05);There is no significant difference in fasting blood glucose between the electro-acupuncture group after modeling and after intervention(P>0.05),while the fasting blood glucose value in the model group is significantly higher after 4 weeks than before modeling,the difference is statistically significant(P<0.05).Although there is no significant change in blood glucose after the intervention in the electro-acupuncture group and after modeling,from the model group's blood glucose change trend,within the four weeks after the model,blood glucose continued to rise,indicating that electro-acupuncture intervention had a suppressive effect on the rise of blood glucose.Compares with the model group,the blood glucose increase value of the electro-acupuncture group is significantly less than that of the model group(P<0.05),it shows that electro-acupuncture can effectively control blood sugar of obese rats with type 2 diabetes.The weight gain of the model group and electro-acupuncture group are lower than that of the control group,and the difference is statistically significant(P<0.05).The weight gain of the electro-acupuncture group is lower than that of the model group,indicating that the electro-acupuncture at Zusanli(ST36),Sanyinjiao(SP6),Weiwanxiashu(EX-B3),and Pishu(BL20)points can control the weight change of obese rats with type 2 diabetes.Compares with the model group,the serum leptin level in the electro-acupuncture group decrease,indicating that electro-acupuncture can improve the serum leptin level in type 2 diabetic obese rats.Compares with the control group,the levels of total cholesterol,triglyceride,and high-density lipoprotein cholesterol in the model group are significantly increase(P<0.05);compares with the model group,the level of serum triglyceride in the electro-acupuncture group is significantly reduce,and the difference is statistically significant(P<0.05);total cholesterol,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,and very low-density lipoprotein cholesterol values all decrease,but the difference is not statistically significant(P>0.05).It can be seen that electro-acupuncture can reduce serum lipid levels in obese rats with type 2 diabetes,especially triglyceride levels.Serum insulin levels are lower in the electro-acupuncture group than in the model group,and the difference is statistically significant(P<0.05).The level of C-peptide in the electro-acupuncture group is lower than that in the model group,but the difference is not statistically significant(P>0.05).Comparing the average score shows that the model group is larger than the electro-acupuncture group,and the electro-acupuncture group is larger than the blank group.It shows that electro-acupuncture can reduce the serum insulin level of obese rats with type 2 diabetes,and can reduce the level of C peptide to some extent.Comparison of the insulin resistance index shows that the insulin resistance index of the electro-acupuncture group is lower than that of the model group,and the difference is statistically significant(P<0.05),indicating that electro-acupuncture can alleviate insulin resistance in type 2 diabetic and obese rats.Experiment 2 shows that the phosphorylation level of IRS-1 serine/threonine 307 in the model group is higher than that in the control group and the electro-acupuncture group,and the difference is statistically significant(P<0.05).There is no significant difference between the control group and the electro-acupuncture group(P>0.05),compares with the mean score shows that the control group have the lowest expression of phosphorylation,followed by the electro-acupuncture group,and the model group have the highest expression.It shows that electro-acupuncture can reduce the phosphorylation level of IRS-1 serine/threonine in skeletal muscle of type 2 diabetic obese rats,and reduce its inhibitory effect on IRS-1 and its downstream pathways.Comparing the levels of IRS-1 tyrosine 895 phosphorylation sites in the three groups shows that the model group have the lowest expression of phosphorylation,and the difference is statistically significant compare with the control group and the electro-acupuncture group(P<0.05),there is no significant difference between the control group and the electro-acupuncture group(P>0.05).It shows that electro-acupuncture can increase the expression of IRS-1 tyrosine phosphorylation in rat skeletal muscle,and adjust the balance between IRS-1 phosphorylation to relieve skeletal muscle insulin resistance.Compares with the control group,the expression level of PI3K p85 protein in the model group is significantly higher,and the difference is statistically significant(P<0.05).Compares with the model group,the expression of PI3K p85 protein in the electro-acupuncture group is increased,and there are significant differences between the two groups(P<0.05),there is no significant difference between the control group and the electro-acupuncture group(P>0.05).This shows that electro-acupuncture can increase the protein expression of PI3K p85 in skeletal muscle of diabetic rats.Compares with the control group and the model group,the expression level of GLUT4 protein in the electro-acupuncture group is significantly increase,and the difference is statistically significant(P<0.05).There is no significant difference between the control group and the model group(P>0.05).The mean score is compared shows that the acupuncture group have the most protein expression,followed by the control group,and the model group had the lowest expression.This shows that electro-acupuncture can increase the expression of glucoprotein in skeletal muscle of rats.[Conclusion]Electro-acupuncture at Zusanli(ST36),Sanyinjiao(SP6),Weiwanxiashu(EX-B3)and Pishu(BL20)can reduce blood glucose and blood lipid levels,reduce insulin and C-peptide levels in type 2 diabetic obese rats,control rat weight and reduce leptin levels,thereby reducing their insulin resistance,alleviate abnormal glucose and lipid metabolism in diabetic and obese rats.At the same time,electro-acupuncture can effectively increase the expression of GLUT4 protein in skeletal muscle,thereby alleviating its insulin resistance.The balance between its mechanism and electro-acupuncture regulating IRS-1 phosphorylation,increasing PI3K protein levels,and reducing its effect on downstream PI3K pathways related. |
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