| Objective: Obesity is a pathological state of energy metabolism disorder caused by excessive or abnormal fat accumulation in the body.Insulin resistance(IR)is the basis of a series of chronic diseases such as dyslipidemia,hypertension,cardiov ascular and cerebrovascular diseases,glucose intolerance,and type 2 diabetes(T2DM).It affects the body’s overall m etabolism.Ingestion of excessively saturated fatty acids in a high-fat diet leads to changes in the intestinal microbial structure,damage to the intestinal mucosal barrier,in creased intestinal permeability,which causes endotoxin(LP S)to enter the bloodstream,and LPS activates through cell membrane receptors Multiple intracellular signal transduct ion pathways cause systemic inflammatory response,promote islet β cell apoptosis,affect insulin signal transduction process,and cause insulin resistance.LPS is considered to be a trigger for obesity and type 2 diabetes.Intestinal barrier damage is not only the result of the deterioration of insulin resistance obesity,but also an important factor to promote the development of insulin resistance.The inte stinal mucosa mechanical barrier formed by intestinal epith elial cells and intercellular connections is the most important barrier among intestinal defensive barriers,and tight junctions are the most important intercellular connection methods.The tight junction protein is an important compone nt of TJ.Among them,Occludin and Z-O1 proteins are the most important tight junction proteins and play an important role in maintaining the connection between intestinal epith elial cells.Early intervention on insulin resistance obesity can prevent it from developing into metabolic diseases s uch as T2 DM.In this experiment,electroacupuncture combined with calorie control is used to intervene in high-fat diet-induced insulin resistance obese model rats,and the changes in body weight,Lee’s index,insulin sensitivity,serum endotoxin content,and small intestinal mucosa tissues、the expression of tight junction protein in the small intestine are observed,to clarify the effect of electroacupuncture combined with calorie control on IR and small intestinal barrier function,and provide an experimental basis for clinical use of electroacupuncture combined with calorie control to treat insulin resistance obesity.Methods: Modeling: A high-fat diet-induced IR obesity model,using body weight,Lee’s index,and GIR as the criteria for evaluati ng the success of modeling.A total of 100 8-week-old SPF Wistar male rats were selected.According to the random number table method,20 rats were taken as normal groups and fe d with normal feed,and the other 80 were used as building blocks for feeding with high-fat feed.After 8 weeks of feeding,the weight and Lee,s index of rats were measured to evaluate the obesity model;We randomly selected 4 rats in t he general feed-fed group and 16 rats in the high-fat diet group,and measured the GIR of the rats with high insulin-positive glucose clamp to evaluate the IR model.Complying with these two evaluation criteria indicates successful creation of an insulin resistance obesity model.Grouping: Sele ct rats that meet the obesity criteria and randomly divide them into model group,electroacupuncture group,calorie control group(heat control group),electroacupuncture combin ed calorie control group(acupuncture heating control group),each group contains 10 rats.Ten rats fed with normal feed were randomly selected into the normal group.There are 5 groups.Intervention methods of each group: normal group: normal rats,fed with normal feed;model group: fed with high-fat feed;electroacupuncture group: fed with high-fat feed,intervention with electroacupuncture;calorie control gr oup: with calorie control intervention(Measure the food int ake of the rats in the model group every day.Taking 70% of the daily food intake of the rats in the model group as the calorie control intervention.And feeding the rats in as ingle cage.Total 8 weeks)Electro-acupuncture combined with heat control group: intervention with electro-acupuncture and heat control.Electroacupuncture intervention method: A cupoint selection: Zusanli,Fenglong,Guanyuan,Zhongwan;Use 0.20 × 13 mm disposable milliseconds,connect to the ele ctrode of HANS LH202 H electroacupuncture instrument,parame ter setting: 2Hz,1m A,continuous wave.10 minutes of elect roacupuncture every day and 3 treatments per week.The total course of treatment is 8 weeks.Index detection: measure body weight,anal and nose length at weeks 0,2,4,6,and 8 of the intervention,and calculate the Lee’s index;at the 6th week of the intervention,me asure the intraperitoneal glucose tolerance and intraperito neal insulin tolerance of all rats;after treatment,3 rats were randomly selected from each group for high insulin-positive glucose clamping to detect GIR,the remaining rats were tested for serum insulin and endotoxin by enzyme-linked immunosorbent assay,and HE staining was used to detect the pathological status of intestinal mucosa in small intestine tissue Changes,Western blot method was used to detect the expression of tight junction protein Occludin in small int estine tissue,and immunohistochemistry was used to detect the expression of tight junction protein Occludin and ZO-1 protein in small intestine tissue.Results: 1.Effect on body weight and degree of obesity: Compared with the model group,after 4 weeks,the weight of rats in the electroacupuncture combined heat control group decreased significantly(P <0.01);at 6 weeks,the weights and models of the three intervention groups The trend decreased compared with the group(P <0.01);after the 8th week of intervention,the weight of the rats in the electroacupuncture combined heat control group was lower than that in the electro acupuncture group and the heat control group(P <0.01).Le e,s index changes are synchronized with body weight.2.The effect on insulin sensitivity:(1)The effect on intraperitoneal glucose tolerance: After intraperitoneal glucose injection for 15 minutes,all rats’ blood glucose increased rapidly;the blood glucose values of the three intervention groups from 30 minutes to 120 minutes were lower than that of the model group(P<0.01).After 30 minutes,the blood glucose value of the electroacupunc ture combined heat control group was lower than that of the electroacupuncture group and the thermal control group(P<0.01);the difference between the electroacupuncture group and the thermal control group was not statistically signifi cant.(2)Effect on intraperitoneal insulin tolerance: compared with the normal group,the model group rats were observed from the 15 th minute to the 120 th minute,and the blood glucose values were higher than the normal group(P <0.01);compared with the model group,three Rats in the intervention group had lower blood glucose values from 15 min to 120 min(P <0.01);compared with the electroacupuncture group,the blood glucose values of the electroacupuncture combined heat control group at 30 and 60 min were lower(P <0.05)Compared with the thermal control group,the blood glucose value of the electroacupuncture combined thermal control group w as lower at 60 min(P <0.05);the difference between the el ectroacupuncture group and the thermal control group was no t statistically significant.(3)Effect on GIR: After 8 weeks of intervention,compared with the normal group,the GIR of the model group was signi ficantly reduced(P <0.01);compared with the model group,the GIR of the three intervention groups was significantly increased(P <0.01);Compared with the electro-acupuncture group and the thermal control group,the GIR of the electr o-acupuncture combined thermal control group decreased(P <0.05);there was no statistical difference between the electro-acupuncture group and the thermal control group.(4)Effect on serum insulin: After 8 weeks of intervention,compared with the normal group,the serum INS content of insulin resistance obese rats in the model group was significantly increased(P <0.01);compared with the model group,the electroacupuncture group,Serum INS content of insulin resistance obese rats in thermal control group and electroacupuncture combined with thermal control group was significantly decreased(P <0.01);compared with electroacupuncture group and thermal control group,serum INS in electroacupuncture combined with thermal control group insulin resistance obese rats The content decreased(P <0.05);there was no statistical difference between the electroacupuncture group and the thermal control group.3.Effect on serum LPS: After 8 weeks of intervention,compared with the normal group,the serum LPS content of insulin resistance obese rats in the model group was significantly increased(P <0.01);compared with the model group,three intervention groups The serum LPS content of obese rats with insulin resistance was significantly reduced(P <0.01);compared with the electroacupuncture group and the thermal control group,the serum LPS content of the insulin resistance obese rats with the electroacupuncture combined with th e thermal control group was more significantly reduced(P <0.05);There was no statistical difference between the electroacupuncture group and the thermal control group.4.Effect on the state of small intestinal mucosa: the three intervention groups had less intestinal villous destruction than the model group,and the Chiu’s score was lower(p <0.05).The electroacupuncture combined with heat control group had the lowest Chiu’s score and the most intestinal mucosa destruction.Light;there was no statistical differenc e between the electroacupuncture group and the thermal cont rol group.5.The effect on tight junction protein of small intestine tissue: The protein expression of small intestine Occludin and ZO-1 in model group was significantly lower than that in normal group(P <0.01).Compared with model group,electr oacupuncture,thermal control,electroacupuncture combined The protein expressions of Occludin and ZO-1 in the thermal control group were up-regulated(P <0.01).The protein exp ression levels of Occludin and ZO-1 in the electroacupuncture combined thermal control group were higher than those in the electroacupuncture group and the thermal control group(P <0.01).Compared with the thermal control group,the protein expression of Occludin and ZO-1 in the electroacupuncture group was higher(P <0.05).Conclusions: 1.Calorie control,electroacupuncture,electroacupuncture combined with calorie control can all reduce the weight and obesity of insulin-resistant obeserats,enhance insulin sensitivity,protect intestinal mucosa,up-regulate the expression of tight junction proteins Occludin and ZO-1 in small intestine tissue,and maintain Intestinal barrier function.2.The combined intervention of electro-acupuncture and calorie control is significantly better than single thermal control intervention and single electro-acupuncture intervent ion in reducing body weight,reducing its obesity,improving insulin sensitivity,reducing serum endotoxin levels,and maintaining small intestinal barrier function.Calorie control intervention has played a synergistic role in the abov e aspects.3.Electroacupuncture and calorie control may play a role in improving insulin sensitivity by improving intestinal bar rierfunction,reducing endotoxin release,and reducing int estinal inflammation.This is a possible mechanism by which electroacupuncture and calorie control exert a synergistic effect to improve insulin resistance in obesity.4.This experiment observes that the effect of electroacupu ncture combined with heat control on the intestinal barrier function of insulin resistant obese rats is limited to the mechanical barrier.The chemical barrier,immune barrier and biological barrier could not be observed in this experiment.The interaction of each barrier function is inseparable.In the future work,we can further observe and study the effect of electroacupuncture combined with heat control on other barrier functions and the relationship between insul in resistance and obesity. |
