| Objectives Obesity is a pathological state in which the body's energy metabolism disorder is caused by multiple factors,resulting in an amount of adipose tissue exceeding the average level and affecting human health.When the body is obese,adipose tissue produces excessive non-esterified fatty acids,adipokines,pro-inflammatory factors,which are closely related to chronic mild inflammation,which promotes insulin resistance and leads to diseases such as type 2 diabetes.Insulin resistance is also a common pathological basis for type 2 diabetes,polycystic ovary syndrome,metabolic syndrome,and hypertension.Therefore,early intervention on obesity and insulin resistance can effectively prevent the occurrence and development of metabolic diseases such as type 2 diabetes.In this study we will intervene high-fat-diet-induced IR obese rat model by “The Combination of Biao-Ben Acupoints” acupuncture.Molecular biologic methods will be used to measure the expression.The aim of the study is to investigate the possible mechanism of acupuncture in modulating chronic inflammation and IR through SIRT1-mediated anti-inflammation pathway and explore the related epigenetic regulation of acupuncture.The project will disclose the important mechanism of acupuncture in improving IR and chronic inflammtion,providing new proof for clinical application of acupuncture in the treatment of IR obesity and the related changes of SIRT1-mediated anti-inflammtion moleculars and the downstream pre-inflammatory genes transcription andMethods Ninety male Wistar rats were selected and firstly adapted for one week.They were randomly divided into normal group(15 rats)and obese group(75 rats),which were given normal feed and high-fat diet.After 8 weeks,3 rats in the normal group and 12 patients in the obese group participate in Andres clamp technique to evaluate the insulin resistance model.The weight and anal length of each rat were measured and calculated the Lee's index,an assessment of obesity models.Those who met the insulin resistance model and the obesity model at the same time were identified as successful modeling.After that,the rats in the obese group were randomly divided into model group?EA group?EA-inhibitor group and CR group,10 rats in each group were fed with high-fat diet;10 samples were randomly selected from the normal group.The feed intake of the model group rats was weighed daily,and after the mean value was calculated,the corresponding food intake of the model group was controlled according to 30% of the heat of the model group.EA group was given electroacupuncture treatment.Acupoints: Zusanli,Fenglong,Guanyuan,Zhongyu,according to the depth of acupuncture3-5mm,frequency 2Hz/1m A,3 times a week,each time 10min intervention treatment,the total course of treatment is 8 weeks;the EA+inhibitor group was given electroacupuncture treatment,and SIRT1 specific inhibitor was intraperitoneally injected.All rats were weighed(baseline)before the intervention,and all rats were measured for body weight every two weeks after the start of the intervention.At the 6th week,IPGTT and IPITT were detected in each group.Intervention at 8thweek,3 rats were randomly selected from each group for Andres clamp technique and glucose infusion rate(GIR)levels were measured.After 8 weeks of intervention,apical blood samples were taken to detect serum inflammatory markers CRP,IL-6 and serum insulin(INS)levels in each group;white adipose tissue was removed and SIRT1 and AP-1 were detected by Western blotting(c-Jun),acetylated AP-1(c-jun)and IL-6 protein expression;real-time fluorescent quantitative PCR was used to detect the expression of downstream inflammatory factor IL-6 m RNA.Results1.Effects of high-fat diet for 8 weeks on body weight,Lee's index and GIR in rats After 8 weeks of feeding,the body weight of the rats in the obese group was significantly higher than that in the normal group(P<0.01).In the Lee's index,the rats in the obese group were higher than the normal group(P<0.05).In the GIR group,the rats in the obese group The level was significantly lower than the normal group(P < 0.01).2.Effects of electroacupuncture on body weight,serum inflammatory factors,serum insulin and insulin sensitivity in insulin resistant obese rats In terms of body weight,after 8 weeks of intervention,the body weight of the EA group and the CR control group was significantly lower than that of the model group(P<0.01),and the body weight of the EA-inhibitor group was lower than that of the model group(P<0.05).Compared with the model group,the CRP content of the EA group was decreased(P<0.05),while the CRP content of the CR group was significantly lower(P<0.01).Compared with the EA group,the needle was compared with the model group.There was no significant difference in CRP content between the EA-inhibitor and the CR control group(P>0.05).Compared with the model group,the IL-6content in the EA group and the CR group were significantly lower(P<0.01).Compared with the EA group,the IL-6 content in the EA-inhibitor was significantly increased(P<0.01).There was no significant difference between the CR group and the EA group(P>0.05).In terms of serum insulin,compared with the model group,the INS content of the EA group and the CR group were significantly decreased(P<0.01),while the content of the EA-inhibitor group was also decreased(P<0.05);Compared with the EA group,the content of INS in the EA-inhibitor increased significantly(P<0.01),while the difference between the CR group and the EA group was not statistically significant(P>0.05).In terms of IPGTT,the abdominal glucose tolerance of the model group was impaired compared with the normal group(P<0.05).Compared with the model group,the glucose tolerance in the EA-inhibitor and the CR group was improved(P<0.05).In the EA group, the glucose tolerance in the abdominal cavity was significantly improved(P<0.01).In the IPITT group,the intraperitoneal insulin tolerance was significantly impaired in the model group compared with the normal group(P<0.01).Compared with the model group,the CR control group was intraperitoneal insulin.The tolerance was improved(P<0.05),the EA group was significantly improved(P<0.01),and the EA-inhibitor group was not statistically different(P>0.05).In terms of GIR,compared with the normal group,The GIR values ??of the model group were significantly lower(P<0.01).Compared with the model group,the GIR values ? ? of the EA group,the EA-inhibitor group and the CR control group were significantly increased(P<0.01).There was a significant difference in the GIR values ??between the EA group and the EA-inhibitor group(P<0.01).The difference in GIR between the EA group and the CR control group was statistically significant(P<0.05).3.Effects of electroacupuncture on the expression of SIRT1/AP-1 and its downstream inflammatory factors in insulin resistant obese rats In terms of SIRT1 protein expression,SIRT1 protein expression was significantly decreased in the insulin resistance obese model group compared with the normal group(P<0.01).Compared with the model group,the SIRT1 protein expression in the EA group and the CR control group was significantly increased(P<0.01);compared with the EA group,SIRT1 protein expression was significantly decreased in the EA-inhibitor group(P<0.01).In the expression of acetylated AP-1 protein,the expression of Ac-c-jun protein in the insulin resistance obesity model group was significantly higher than that in the normal group(P<0.01);compared with the model group,the expression of Ac-c-jun protein in the EA group,the EA-inhibitor group and the CR control group was significantly decreased(P<0.01).Compared with the EA group,the expression of Ac-c-jun protein in the CR control group was significantly decreased.(P<0.01),while the expression of Ac-c-jun protein in the acupuncture plus group was increased(P<0.05).In terms of IL-6 protein expression,compared with the normal group,the IL-6 protein level of the insulin resistance obese model group was significantly increased(P<0.01);compared with the model group,the EA group and the EA-inhibitor group The protein level of IL-6 in the CR group was significantly decreased(P<0.01).Compared with the EA group,the IL-6 protein in the EA-inhibitor group was significantly increased(P<0.01).Conclusions1.Electroacupuncture can inhibit the body weight of obese insulin resistance rats induced by high-fat diet,reduce the level of serum inflammatory factors,and improve the chronic inflammatory state of the body to some extent.At the same time,electroacupuncture can enhance insulin sensitivity and improve the body of obese insulin resistance rats.The state of insulin resistance.2.Electroacupuncture can up-regulate the expression of SIRT1 protein in obese insulin-resistant rats,which reduces the protein expression of acetylated AP-1,which leads to a corresponding decrease in protein expression and m RNA expression of downstream inflammatory factor IL-6. |
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