| [Significance]Diabetes is a chronic abnormal lipid metabolism disease characterized by elevated blood glucose.The pathological basis is mainly insulin deficiency and insulin resistance,which are caused by multiple factors such as genetics and environment.Type 2 diabetes accounts for about 95%of the total number of people with diabetes,"Glucose Depot" The liver is the main target organ for insulin,and it is an important place for glucose metabolism and various other metabolisms.Strengthening the improvement of liver function,enhancing the receptor activity on the liver cell membrane surface,and improving liver damage can significantly regulate insulin secretion and insulin resistance.In addition,since the current clinical western medicines have certain side effects and adverse reactions,there are a large number of documents confirming that electroacupuncture has a significant improvement effect on type 2 diabetes.Therefore,this experiment aims to provide theoretical research on the mechanism of electroacupuncture to lower fasting blood glucose and improve insulin resistance in liver tissue from the perspective of liver tissue.[Objective]1.To observe the effects of electroacupuncture on the body weight,fasting blood glucose,insulin,C-peptide levels,blood lipids and liver-related transaminase in type 2 diabetic rats.HE staining to observe the changes of liver morphology,through comparing three interventions:blank group,model group,and electroacupuncture group,to observe whether electroacupuncture can reduce fasting blood glucose and blood lipid levels to protect liver tissue,and verify whether electroacupuncture can effectively improve insulin resistance in type 2 diabetes.2.Explore whether electroacupuncture can activate the adenylate-activated protein kinase AMPK(AMP-activated protein kinase,AMPK)/Peroxisome Proliferators-activated receptor-ycoactivator-1α(Peroxisome Proliferators-activated receptor-ycoactivator-1α,PGC-1α)signaling pathway and induce the conversion of adenosine triphosphate(ATP)and adenosine monophosphate(AMP)energy,which in turn up-regulates the expression of Glucose transporter 4(GLUT4)protein in liver tissue,to reduce fasting blood glucose and improve insulin resistance.Western Blot method was used to observe the regulation of AMPK,PGC-1α and GLUT4 levels in liver tissue after electroacupuncture intervention,and the conversion of ATP and AMP energy in liver tissue was detected by Elisa method,so as to explore the mechanism of electroacupuncture intervention in type 2 diabetes.[Method]Eight 2-month-old SPF-grade male ZL(Zucker Lean)rats(fa/+)were selected as the blank group,and 16 SPF-grade male ZDF(Zucker Diabetic Fatty)rats(fa/fa)at the same month-old age were selected as experimental subjects.Advanced adaptive feeding was carried out for 1 week,and then 24 rats were fed with high-fat and high-sugar induced feed for 4 weeks,with free diet and water.After induction for 4 weeks,the fasting blood glucose and OGTT 2 h blood glucose were both>11.1 mmol/L as the modeling standard.The 16 ZDF rats were randomly divided into model group and electroacupuncture group,each with 8 rats;the blank group and model group were not subjected to any intervention except the same grasping and fixation as the electroacupuncture group.In the electroacupuncture group,acupuncture on both sides of Pishu,Weiwanxiashu,Zusanli and Sanyinjiao,with a direct penetration depth of about 7 mm.One end is connected to Zusanli,and the other end is connected to the same side of Weiwanxiashu.Change to the other side every other day.Electroacupuncture,adjusting frequency 15 Hz,current intensity about 4-6 mA(just keep the rat’s limbs slightly shaking)keep the needle for 20 min,6 times a week(Monday to Saturday),continuous intervention for 4 weeks.The body weight and fasting blood glucose(FBG)were measured before the intervention and after the last intervention.After the last intervention,the serum was collected,and various blood lipid indexes were detected by colorimetry;the liver tissue-related transaminase(Alanine transaminase,ALT),aspartate transaminase(AST)and y-glutamyl transpeptidase(GGT)enzymatic index levels;observe the level of insulin and C-peptide.Part of the rat liver tissues were taken in the ice bath state,and HE staining was performed to observe the morphological changes of liver cells.Western Blot method was used to detect the protein expression levels of AMPK,PGC-1α and GLUT4 in liver tissue;the ATP and AMP content levels were detected by Elisa method.[Result]Experiment 1:Statistics show that before intervention,compared with the blank group,the model group has a higher body weight,and the difference is very statistically significant(P<0.01).Compared with the model group,the electroacupuncture group has a higher body weight.There is almost no difference with the model group,and the difference is not statistically significant(P>0.05),indicating that it is comparable.After intervention,the weight gain of the model group was lower than that of the blank group,and the difference was statistically significant(P<0.01).The weight gain of the electroacupuncture group was lower than that of the model group,but the difference was not statistically significant(P>0.05),but comparing the mean weight after intervention showed that the electroacupuncture group can control the weight change trend of type 2 diabetes insulin resistance rats,and inhibit the growth rate of rat body weight.Before intervention,the fasting blood glucose of the model group was significantly higher than that of the blank group,and the difference was statistically significant(P<0.05).Compared with the model group,the fasting blood glucose of the electroacupuncture group was not statistically significant(P>0.05),and it was comparable.After intervention,the fasting blood glucose of the model group was significantly different from that of the blank group(P<0.01).Compared with the model group,the fasting blood glucose of the electroacupuncture group was significantly lower,and the difference was very statistically significant(P<0.01).).Electroacupuncture can effectively reduce fasting blood glucose and improve insulin resistance.After the intervention,the electroacupuncture group was compared with the model group’s total cholesterol(TC),high density lipoprotein(HDL),low density lipoprotein(LDL),and very low density lipoprotein(vLDL)decreased significantly(P<0.05),and the difference was statistically significant.Among them,LDL and vLDL decreased significantly(P<0.01).The level of insulin in the electroacupuncture group was significantly lower than that of the model group,and the difference was statistically significant(P<0.01).The C-P level of the electroacupuncture group was lower than that of the model group,and the difference was statistically significant(P<0.05);before and after intervention,The area under the OGTT curve of the model group was higher than that of the blank group,and the difference was statistically significant(P<0.05).Before intervention,the area under the OGTT curve of the electroacupuncture group was not statistically different from the model group(P>0.05),indicating that it was comparable.After intervention,the area under the OGTT curve of the electroacupuncture group was significantly decreased compared with the model group,and the difference was statistically significant(P<0.05).Electroacupuncture can effectively improve the serum levels of insulin INS,C-P and glucose tolerance in type 2 diabetic insulin resistance rats.After intervention,the AST and ALT levels of the electroacupuncture group decreased compared with the model group,but the difference was not statistically significant(P<0.05).The GGT level of the electroacupuncture group decreased significantly,and the difference was statistically significant(P<0.05).Electroacupuncture can adjust the level of related transaminase in the serum of insulin resistance rats with type 2 diabetes,thereby reducing the degree of liver cell damage.HE staining showed that the electroacupuncture group had improved hepatocyte cytoplasm red staining,nuclear pyknosis,cell apoptosis,and vesicular lipid droplets compared with the model group.Experiment 2:After intervention,compared with the blank group,the expression of AMPK and GLUT4 protein in rat liver tissues was significantly reduced in the model group,and the difference was statistically significant(P<0.01),and the expression of PGC-1αprotein in the model group was reduced,and the difference was statistically significant(P<0.05).The protein expression levels of AMPK,PGC-1α and GLUT4 in the electroacupuncture group were higher than those in the model group,and the difference was statistically significant(P<0.05).Compared with the blank group,the levels of ATP and AMP in the liver tissue of the model group were significantly decreased and the difference was statistically significant(P<0.05),the contents of ATP and AMP in the liver tissue of the electroacupuncture group were higher than those of the model group and the difference was statistically significant(P<0.05).It shows that electroacupuncture can activate AMPK/PGC-1a pathway to induce energy changes and up-regulate the expression of GLUT4 protein to improve liver insulin resistance.[Conclusion]Electroacupuncture can reduce fasting blood glucose and blood lipid levels in insulin resistance rats with type 2 diabetes,lower insulin and C-peptide levels,inhibit weight gain in rats,reduce liver-related transaminase levels,and protect liver damage,thereby improving insulin resistance in liver tissues and alleviating type 2 diabetes abnormal blood glucose and lipid metabolism in rats.Electroacupuncture can up-regulate the levels of ATP and AMP in liver cells,thereby improving energy metabolism and reducing insulin resistance in liver cells.The mechanism is related to the regulation of AMPK/PGC-1α pathway activity by electroacupuncture and up-regulating the expression level of GLUT4 protein.This research was successfully completed based on the National Natural Science Foundation general project(Effect of electro-acupuncture in regulation of fast and slow twitch muscle fibers through the AMPK/PGC-1a pathway to improve insulin resistance.No:81973935). |
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