| Objective: To observe the obese insulin resistant Wistar rats that were successfully modeled by using a high-fat diet,through the method of using electric needle combined with calorie control.By observing the regulation and influence of this intervention on Silent Information Regulator 1(Sirt1),nuclear factor ?-B(NF-?B),downstream inflammatory factors,tumor necrosis factor alpha(TNF-?)and the polarization state of Macrophages,which are in white adipose tissue of the abdomen.Through the chronic inflammation of white adipose tissue,to study the effects of electroacupuncture and calorie control on obese insulin resistance rats,in order to provide a corresponding experimental basis for the role of electroacupuncture combined with caloric control in obesity,and its prevention and treatment of insulin resistance and related diseases.Methods:Choose ninety male Wistar rats.After 1 week of normal feed feeding,15 rats were randomly selected to the normal diet group(normal feed)and 75 in the model(high fat diet feed).After8 weeks of feeding,select 5 rats in the normal group randomly,and 25 rats in the model group.Then evaluate the insulin resistance model by high insulin-positive glucose clamp,and measure the body weight and anal length of each rat and the Lee's index to evaluate the obesity model.And the insulin resistance-resistant obese rats are successfully modeled in accordance with the two evaluation criteria.The model rats fed with high-fat diet were randomly divided into five groups: model group,electroacupuncture group,electroacupuncture combined with inhibitor group,calories control group,and electroacupuncture combined with calories control group,with 10 rats in each group.Randomly select ten rats in the normal diet group as the normal group by the same method.After grouping,the normal diet group continued to feed with normal diet,and the remaining groups continued to be fed with high fat diet.Among them,high fat diet feeding the model group without other intervention.Used electroacupuncture treatment for The electroacupuncture group and the acupuncture points were selected as follows: Zhongwan point,Tianshu point on both sides,Guanyuan point,Fenglong point and Zusanli point on both sides,the depth of acupuncture was 3-5mm,and the frequency of electroacupuncture was 2Hz/1m A,10 minutes per treatment,3 treatments per week.Rats in the electroacupuncture combined with inhibitor group group were given intraperitoneal injection of Sirt1 inhibitor nicotinamide three times a week in addition to electroacupuncture treatment.The intervention method of the calorie control group is regularly weighing and recording the feeding amount and remaining amount of the rat in the model group,and reduced by 30% on the basis of their food intake.The electroacupuncture and calories control group gives electro-acupuncture and calories control intervention.The intervention was conducted for a total of 8 weeks.Weighed and record the weights of rats at0,2,4,6,and 8 weeks of intervention;and tested for IPGTT and IPITT at week 6.Eight weeks after the intervention,dissected the rat to extract experimental material.Took the blood of the apex and the content of TG,TC and FFA in serum was detected by Elisa et al.The expression of Sirt1,NF-?B?ac-NF-?B and TNF-? protein was detected by Western blotting.Detected the expression of TNF-?gene by PCR.Detected the morphological changes of adipocytes by HE staining.Used the Immuno-histochemistry to detect the expression of CD206 and CD11 c in macrophages.Results:1.Effects of high-fat diet on body weight,Lee's index and glucose infusion rate(GIR)in rats:Compared with the normal diet group,the body weight of the model rats was significantly increased(P<0.01),the growth rate of Lee's index increased,the difference was significant(P<0.05),and the GIR decreased significantly(P<0.05).2.Effects of electroacupuncture combined with calorie control on serum TC,TG,FFA and insulin sensitivity in insulin resistant obese rats:Compared with the model group,the serum levels of TC,TG and FFA in the intervention group showed a downward trend,and the serum levels of the indicators in the electroacupuncture group decreased(P<0.05).The contents of TC,TG and FFA in the serum of rats were significantly decreased(P<0.01).The each intervention group's IPITT and IPGTT values was lower than model groups(P<0.05),Electroacupuncture combined with calorie control group was significantly lower than model group(P<0.01).3.Effects of electroacupuncture combined with calorie control on Sirt1?NF-?B? acetylated NF-?B and TNF-? in the white adipose tissue of insulin resistant rats:Compared with the content of Sirt1 in the the model group,electroacupuncture group and the calories control group were increased(P<0.05),and the electroacupuncture+ caloric control group was significantly increased(P<0.01);the levels of of NF-?B and degree of acetylation? TNF-?in the electroacupuncture group and the other calories control group were decreased(P<0.05),and the content of TNF-? in electroacupuncture with caloric control group was significantly decreased(P<0.01).4?Effect of Electroacupuncture Combined with Caloric Control on Polarization Status of Macrophages in Rats with Insulin Resistance:Compared with the expression level of CD11 c in the model group,the content of the intervention group showed a downward trend,and the content of electroacupuncture group and thermal control group decreased significantly(P<0.05),and the electroacupuncture combined with calorie control group decreased significantly(P<0.01).Compared with the model group,the content of CD206 increased in the intervention group(P<0.05),and the electroacupuncture combined with the thermal control group showed a significant upward trend(P<0.01).Conclusion:1.Electroacupuncture combined with calorie control can reduce the body weight of insulin resistant obese rats,reduce the content of serum TC,TG,FFA,adjust lipid metabolism,enhance insulin sensitivity and mitigate insulin resistance in rats.2.Electroacupuncture combined with caloric control can increase the expression of CD206 in macrophages,at the same time,reduce the content of CD11 c,suggesting that electroacupuncture combined with caloric control may improve the inflammatory state of adipose tissue by improving the polarization state of macrophages.3.Electroacupuncture combined with calorie control can up-regulate the expression of Sirt1 in white adipose tissue of obese insulin-resistant rats,reduce the acetylated NF-?B's expression,as well as TNF-?,directing that electroacupuncture combined with caloric control may up-regulate Sirt1.Decreased expression of NF-?B and acetylated NF-?B,inhibits NF-?B transcriptional activity,reduces the expression of downstream inflammatory factors such as TNF-?,Thereby affecting the polarization of macrophages,reducing the inflammatory state of adipose tissue,controlling obesity,enhancing insulin sensitivity. |
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