| Objectives The incidence of obesity is increasing recent years.Adipose tissue not only deposits fat,but also secretes many inflammatory cytokines,leading to insulin resistance by activating inflammatory signaling pathway.Insulin resistance is the common pathological basis of various diseases such as type 2 diabetes.Thence,on the basis of previous studies,this experiment took insulin-resistant obese rats as a model,breaking in from the nuclear factor κB(NF-κB)signaling pathway mediated by silent information regulator 1.To observe the effect of electroacupuncture(EA)on inflammatory factors and insulin sensitivity in adipose tissue of insulin-resistant obese rats,and to further explore the effect of histone acetylation modification induced by deacetylation of SIRT1 on the transcription of NF-κB signaling pathway.To investigate the epigenetic mechanism of EA in improving obesity and insulin resistance by controlling adipose tissue inflammation.Methods One hundred and twenty 8-week-old SPF Wistar male rats were raised.Thirteen rats were randomly selected as normal(NOR)group and fed with normal diet.The rest were fed with high-fat diet for modeling.After 8 weeks,65 rats which reached the obesity standard were divided into model(MOD)group,EA group,sham acupoint(SA)group,electroacupuncture combined inhibitor(EI)group and resveratrol(RSV)group,with 13 rats in each group.Three rats in each group were randomly selected for insulin sensitivity detection by hyperinsulinemia-euglycemic clamp.Then the corresponding intervention treatment was given to each group.(1)NOR group: fed with normal diet without other intervention;(2)MOD group: fed with high-fat diet without other intervention;(3)EA group: Zusanli(ST36),Fenglong(ST40),Zhongwan(CV12)and Guanyuan(CV4)with a EA frequency at 2Hz,1m A,continuous wave connected with Korean EA therapeutic apparatus.15 minutes treated by EA once time,three times a week,a total of 8 weeks of treatment.(4)SA group: the acupoint were taken about 5mm away from the acupionts in EA group and the electrode was clamped,without electrification.Treatment time and frequency are the same as EA.(5)RSV group: resveratrol solution was given intragastrically per 200mg/kg according to the body weight of rats.Three times a week for 8 weeks.(6)EI group: the therapeutic scheme of EA was the same as that of EA group.In addition,sirtinol inhibitor solution was injected intravenously into the tail vein per 1mg/kg according to the body weight of the rats.The course of treatment was 3 times a week for 8 weeks.During the intervention,the body weight,anal to nasal length of rats were measured at 0,2,4,6 and 8 weeks,and Lee’s index was calculated.At the 6th week of intervention,the levels of intraperitoneal glucose tolerance test(IPGTT)and intraperitoneal insulin tolerance test(IPITT)were measured in each group.At the end of the intervention,3 rats in each group were randomly selected for hyperinsulin-normal glucose clamp to detect systemic insulin sensitivity.After the rats were executed,fresh tissue and the formaldehyde perfusion tissue were taken for detection respectively.(1)Enzyme-linked immunosorbent assay(ELISA): The levels of serum CRP,IL-6,TNF-α and insulin were measured in blood collected from rat heart before executed.(2)Western blotting(WB): the protein expressions of SIRT1,IL-6,TNF-α,acetylated NF-κB(Ac-NFκB),NF-κB,acetylated H3K9(Ac-H3K9),and H3 were detected in rat adipose tissue.(3)Real-time quantitative PCR(RT-PCR): RT-PCR was used to detect the gene expression of SIRT1,IL-6 and TNF-α in adipose tissue.(4)Immunohistochemical method: Immunohistochemistry was used to detect the infiltration of macrophages CD68 in adipose tissue and the polarization status of M1 macrophages(CD11c)and M2 macrophages(CD206).(5)Immunofluorescence double labeling method: the co-expression of SIRT1/Ac-NFκB and SIRT1/Ac-H3K9 in adipocytes was detected.(6)Chromatin immunoprecipitation(CHIP): The degree of acetylation of H3K9 in the promoter region of NF-κB gene in NOR group,MOD group and EA group was verified.Statistical analysis was performed after the detection was completed.Results 1.EA can reduce the body weight,serum inflammatory factor levels and improve insulin sensitivity in insulin-resistant obese rats.(1)Body weight results showed that except for the EA group and RSV group,the body weight of rats in other groups showed an upward trend during the intervention.Before intervention(ie,at the 0th week,after the modeling was completed),the weight of the other groups was significantly higher than that of the NOR group(P<0.01),and more than 20% of the mean weight of the NOR group,suggesting that the obese rats induced by high-fat diet were successful.Compared with the model group,the body weight of the RSV group began to decrease from the 4th week of intervention(P<0.05),which suggested that the activation of SIRT1 could reduce the body weight of obese rats.From the 6th week of intervention,the body weight of EA group and RSV group decreased visibly(P<0.01).This indicated that EA had the same effect as RSV.However,there was no significant difference between the SA group and the MOD group.This suggested that non-acupoint acupuncture did not improve the body weight of insulin-resistant obese rats.There was no significant difference between the EI group and the MOD group,but the average body weight of the EI group was lower than that of the MOD group and higher than that of the EA group,indicating that the SIRT1 inhibitor blocked the effect of EA.The down-regulation effect of EA on body weight is related to the activation of SIRT1.By the 8th week of intervention,the trend of EA group,SA group,EI group and RSV group was consistent with the 6th week compared with the MOD group.This suggests that EA and RSV had a stable effect on weight loss.(2)The results of Lee’s index showed that before the intervention,the Lee’s index of the other groups was markedly increased compared with the NOR group(P<0.01),suggesting that rats with high-fat diet showed obesity.Compared to the MOD group,the Lee’s index of the EA group and the RSV group significantly decrease from the 6th week of intervention(P<0.05,P<0.01).This indicates that an increase in SIRT1 can reduce the obese state of the rat,and the EA has the same effect as the RSV.There was no obviously difference between the SA group and the MOD group,which confirmed that EA points rather than non-acupoint acupuncture can reduce the obesity of rats.The same result appeared between the comparison of the needle suppression group and the model group.It is indicated that SIRT1 inhibitors block the effect of EA,and the effect of EA on reducing Lee’s index is related to the activation of SIRT1.At the 8th week of intervention,the trend of EA group,SA group,EI group,RSV group and MOD group was consistent with the trend of the 6th week,reaffirming that EA and RSV can stably reduce obesity.(3)IPGTT experiments showed that there was no statistical difference in basal blood glucose between the groups in the fasting state.Compared with the NOR group,the blood glucose of the MOD group rats was clearly higher than that of the NOR group from the 30 th minute to the 120 th minute(P<0.05),which suggested that the MOD group rats had decreased intraperitoneal glucose tolerance levels.The blood glucose level of the EA group was significantly lower than that of the MOD group(P<0.01)from the 30 th minute to the 120 th minute,suggesting that the EA can improve the insulin resistance of obese and insulin resistant rats.However,there is no such effect in the SA group.There was no statistical difference in blood glucose values between the EA group and the MOD group.This indicates that non-acupoint acupuncture treatment does not improve the abdominal glucose tolerance in insulin resistant obese rats. There was no difference between the EI group and the MOD group at the 30 th minute.However,from the 60 th minute to the 120 th minute,the blood glucose level was visibly lower than that of the MOD group(P<0.01),but significantly higher than that of the EA group.It indicated that the SIRT1 inhibitor block the role of EA to some extent,and the effect of EA on the improvement of glucose tolerance in the abdominal cavity was related to SIRT1.In the RSV group,the blood glucose level was significantly lower than those in the MOD group from the 15 th minute until the 120 th minute(P<0.01).This suggests that up-regulation of SIRT1 can increase the level of abdominal glucose tolerance in insulin-resistant obese rats.(4)IPITT results showed that there was no statistical difference in basal blood glucose between the groups in the fasting state.Compared with the NOR group,the blood glucose of the MOD group was significantly increased from the 15 th minute to the 120 th minute(P<0.05).This suggests a decrease in the intraperitoneal insulin tolerance level in the MOD group rats.The blood glucose levels of rats in the EA group were obviously lower than those in the MOD group from the 15 th minute to the 120 th minute(P<0.05,P<0.01).It is indicated that EA increase the intraperitoneal insulin tolerance level in insulin resistant obese rats.There was no statistical difference between the measurement of the SA group and the MOD group.This suggests that non-acupoint acupuncture treatment can not improve the intraperitoneal insulin tolerance level in insulin resistant obese rats.Similarly,there was no difference between the EI group and the MOD group.However,from the 15 th minute until the 120 th minute,the blood glucose level of EI group was lower than the MOD group and higher than the EA group. This indicates that the SIRT1 inhibitor blocks the action of the EA.The effect of EA on the improvement of insulin tolerance in the abdominal cavity is related to SIRT1.The RSV group had lower blood glucose values than the MOD group,starting from the 15 th minute until the 120 th minute(P<0.01).This indicates that activation of SIRT1 can significantly increase the intraperitoneal insulin tolerance level in insulin resistant obese rats.(5)The GIR results showed that the GIR levels of the rats fed with high-fat diet were significantly lower than those of the rats fed by normal diet before the intervention(ie,8 weeks after model establishment)(P<0.01).However,there was no significant difference between the groups fed with high-fat diet.Visibly,the systemic insulin sensitivity of the obese rats induced by high-fat diet was decreased.The model of insulin-resistant obese rats is successful.Compared with the MOD group,GIR in the EA group was significantly higher than that in the MOD group(P<0.01).These results suggest that EA can improve systemic insulin sensitivity in insulin-resistant obese rats.However,there was no statistical difference between the SA group and the MOD group,which indicated that non-acupoint acupuncture could not change the insulin sensitivity of insulin-resistant obese rats.GIR in the EI group was higher than that in the MOD group(P<0.05),but the value was lower than that in the EA group,indicating that the SIRT1 inhibitor may inhibit the effect of EA in a certain extent.The effect of EA on systemic insulin sensitivity in insulin-resistant obese rats may be related to the activation of SIRT1.GIR levels in the RSV group obviously higher than the MOD group,which indicated that the activation of SIRT1 was helpful to improve the systemic insulin sensitivity of insulin-resistant obese rats.(6)Serum CRP results showed that serum CRP was significantly increased in the MOD group compared with the NOR group(P<0.01),indicating that the inflammatory CRP protein in the blood of insulin resistant obese rats was higher than that in the NOR group.After EA intervention,serum CRP levels decreased significantly(P<0.01).This suggests that EA can effectively alleviate the level of inflammatory CRP in the blood of insulin resistant obese rats.There was no statistical difference between the SA group and the MOD group.Visbily,non-acupoint acupuncture has no effect on reducing blood inflammatory CRP in insulin resistant obese rats.The level of serum CRP in the EI group was lower than that in the MOD group and higher than that in the EA group(P<0.05).These results suggest that SIRT1 inhibitors can block the regulation of EA on inflammatory CRP,and this regulation is associated with SIRT1.Compared with the MOD group,the serum CRP in the agonist group was significantly decreased(P<0.01),which indicated that the activation of SIRT1 could significantly improve the high level of CRP in the blood of insulin-resistant obese rats.(7)The results of serum IL-6 showed that the level of serum IL-6 in the MOD group was significantly higher than that in the NOR group(P<0.01),indicating that inflammatory factors in the blood of insulin-resistant obese rats fed with a high-fat diet IL-6 increased.After EA intervention,serum IL-6 was obviously decreased compared with the MOD group(P<0.01).It is suggested that EA can reduce the level of inflammatory factor IL-6 in the blood of insulin-resistant obese rats.There was no difference between the SA group and the MOD group,indicating that the non-acupoint acupuncture had no anti-inflammatory effect.Serum IL-6 levels in the EI group were not statistically different from those in the MOD group,but their values were lower than those in the MOD group and higher than those in the EA group(P<0.05).These results suggest that SIRT1 inhibitor can significantly block the regulatory effect of EA on serum inflammatory factors in insulin-resistant obese rats.And this effect is related to the activation of SIRT1 by EA.Compared with the MOD group,the serum IL-6 level in the RSV group was apparently lower than that in the MOD group(P<0.01).It is confirmed that activation of SIRT1 improve the level of serum inflammatory factor IL-6 in insulin-resistant obese rats.(8)Serum TNF-α results showed that,in the MOD group,the levels of serum TNF-αwere higher than those in the NOR group(P<0.05).It is suggested that a high-fat diet can cause obesity in rats and increase the expression of inflammatory factors in the blood.Serum TNF-α was significantly decreased after an EA intervention(P<0.01).This shows that EA can reduce the level of inflammatory factor TNF-α in the blood of insulin-resistant obese rats.There was no apparent difference between the SA group and the MOD group,indicating that the non-acupoint acupuncture had no anti-inflammatory effect.Serum TNF-α in the EI group was lower than that in the MOD group(P<0.05).However,the value was still higher than that in the EA group.It is suggested that SIRT1 inhibitor can reduce the regulation function of EA to a certain extent,and this function of EA may be related to the activation of SIRT1.The level of serum TNF-α in the RSV group was obviously lower than that in the MOD group(P<0.01).It was confirmed that the activation of SIRT1 can improve the level of serum inflammatory factor TNF-α in insulin resistant obese rats.2.EA increase the expression of SIRT1 in adipose tissue of insulin resistant obese rats and decrease the expression of IL-6,TNF-α and macrophage infiltration.(1)The expression of SIRT1 showed that the protein and m RNA expression of SIRT1 in the adipose tissue of the MOD group were significantly lower than those in the NOR group(P<0.05,P<0.01).This suggests that the expression of SIRT1 is decreased in adipose tissue of insulin resistant obese rats.After EA intervention,the protein expression and m RNA expression of SIRT1 increased apparently(P<0.05).It can be seen that the EA activate the expression of SIRT1.There was no statistical difference between the SA group and the MOD group,indicating that non-acupoint acupuncture had no effect on the expression of SIRT1,which further confirmed the efficacy of EA.The protein and m RNA expression of SIRT1 in the RSV group was visibly higher than that in the MOD group(P<0.01).This suggested that resveratrol can effectively activate the expression of SIRT1 in adipose tissue.There was no significant difference in the expression of SIRT1 protein in adipose tissue between EI group and MOD group.Although the m RNA expression of SIRT1 was significantly decreased(P<0.05)in EI group compared to MOD group,there was no difference between the EI group and the EA group also.This indicates that the SIRT1 inhibitor partially blocked the effect of the activation of SIRT1 by EA.(2)The expression of IL-6 showed that the protein expression and m RNA expression of IL-6 in the adipose tissue of the MOD group were significantly higher than those in the NOR group(P<0.01).The results showed that the adipose tissue of insulin-resistant obese rats was in an inflammatory state.Compared with the MOD group,the protein expression and m RNA expression of IL-6 were apparently decreased(P<0.01).It can be seen that EA can control the expression of inflammatory factor IL-6 in adipose tissue.There was no statistical difference between the SA group and the MOD group,indicating that non-acupoint acupuncture did not rduce the effect of inflammatory factor IL-6 in adipose tissue,which further affirmed the anti-inflammatory effect of EA.The protein expression and m RNA expression of IL-6 in the RSV group were obviously lower than those in the MOD group(P<0.01).Visibly,the activation of SIRT1 can effectively control the inflammation of adipose tissue.The expression of IL-6 protein and m RNA in adipose tissue of EI group was significantly lower than that of MOD group(P<0.05),but higher than that of EA group(P<0.05).This indicated that SIRT1 inhibitor partially blocked the anti-inflammatory effect of EA.(3)The results of TNF-α expression showed that the TNF-α protein and m RNA expression in adipose tissue of the MOD group were obviously higher than those of the NOR group(P<0.01),indicating that the adipose tissue of insulin resistant obese rats was in an inflammatory state.After EA intervention,the expression of TNF-α protein and m RNA decreased significantly(P<0.01).These results suggest that EA decrease the expression of inflammatory factor TNF-α in adipose tissue.However,there was no significant difference between the SA group and the MOD group.This indicates that non-acupoints acupuncture has no effect on reducing inflammation of adipose tissue.The protein and m RNA expression of TNF-α in RSV group were significantly lower than those of MOD group(P<0.01).Visibly,the activation of SIRT1 effectively control the expression of inflammatory factors in adipose tissue,which also confirms the anti-inflammatory effect of SIRT1.Compared with the MOD group,the protein and m RNA expression of TNF-α in adipose tissue in the EI group were decreased(P<0.05),but the protein expression of TNF-α was higher than that in the EA group(P<0.05).This indicates that SIRT1 inhibitor block the anti-inflammatory effect of EA to a certain extent.(4)Macrophage CD68 infiltration results: There were different degrees of macrophage infiltration in adipose tissue of rats in each group.The macrophage infiltration around the adipocytes was relatively increased in the MOD group,the SA group,and the EI,while the NOR group,the EA group,and the RSV group were relatively less infiltrated.The results of semi-quantitative analysis showed that the expression of CD68 in macrophages in the MOD group was raised compared with the NOR group(P<0.01).Compared with the MOD group,the expression of CD68 in EA group decreased obviously(P<0.05).This suggests that EA can control macrophage infiltration in adipose tissue.There was no statistical difference between the SA group and the MOD group,indicating that non-acupoint acupuncture could not improve the infiltration of macrophages in adipose tissue.The expression of CD68 in macrophages in RSV group was significantly lower than that in MOD group(P<0.01).These results suggest that the activation of SIRT1 effectively reduce the infiltration of macrophages in adipose tissue.The expression of CD68 in EI group was not different compared to the MOD group and the EA group.These results suggest that SIRT1 inhibitor partially block the effect of EA and increase the infiltration of macrophage CD68 in adipose tissue of insulin-resistant obese rats after EA intervention.It can be seen that the control of EA on macrophage infiltration in adipose tissue of obese rats with insulin resistance may be related to the activation of SIRT1 by EA.(5)The polarization of macrophages showed that M1 macrophages were labeled with CD11 c and M2 macrophages were labeled with CD206.The infiltration of CD11 c macrophages in adipose tissue of MOD group and SA group was relatively increased,while that of EA group and RSV group was relatively decreased.The infiltration of macrophage CD206 in adipose tissue of EA group and RSV group was relatively increased,while that of MOD group and SA group was relatively decreased.From the M1/M2 ratio,the ratio of M1/M2 in the MOD group was significantly higher than that in the NOR grou(P<0.01),which suggested that M1 macrophage infiltration was dominant in adipose tissue of insulin-resistant obese rats.After EA intervention,the ratio of EA group decreased significantly(P<0.01).The results showed that after EA intervention,M1 macrophage infiltration decreased and M2 macrophage increased in adipose tissue of insulin-resistant obese rats.However,there was no significant difference in the ratio between the SA group and the MOD group,suggesting that M1 macrophage infiltration was dominant in the adipose tissue of the SA group,and the relative contents of M1 and M2 macrophages were not adjusted by non-acupoint acupuncture treatment.The ratio of RSV group was obviously lower than that of MOD group(P<0.01),which indicated that RSV intervention decreased M1 macrophage infiltration and increased M2 macrophage content in adipose tissue.These results suggest that the activation of SIRT1 can effectively promote the transformation of M1 macrophages into M2 macrophages in adipose tissue of insulin-resistant obese rats. The ratio of EI group was lower than that of MOD group(P<0.01),but significantly higher than that of EA group(P<0.01).It can be seen that SIRT1 inhibitor blocked the effect of EA to a certain extent.It is indicated that EA can decrease the infiltration of M1 macrophages in adipose tissue and the increase of M2 macrophages is related to the activation of SIRT1 by EA.3.EA reduces the expression of acetylated NF-κB in adipose tissue by activating SIRT1 and co-localization of SIRT1/Ac-NFκB exists in the nucleus.(1)The expression of Ac-NFκB protein: compared with the NOR group,the proportion of Ac-NFκB in the total NF-κB in adipose tissue of the MOD group was significantly raised(P<0.05).This directly proves that the chronic inflammatory of insulin-resistant obese rats increase the probability of acetylation of NF-κB protein in adipose tissue.After EA intervention,the proportion of Ac-NFκB in the total amount of NF-κB protein in adipose tissue decreased significantly(P<0.05).It can be seen that EA effectively reduce the acetylation of NF-κB in adipose tissue.There was no significant difference between the SA group and the MOD group,indicating that non-acupoint acupuncture had no effect on the protein expression of acetylated NF-κB in adipose tissue.Compared with the MOD group,the ratio of Ac-NFκB in the total amount of NF-κB protein in the RSV group was significantly decreased(P<0.01),indicating that activation of SIRT1 apparently reduce the acetylation of NF-κB in adipose tissue of insulin-resistant obese rats.There was no statistical difference in the proportion of Ac-NFκB in the total amount of NF-κB between the EI group and the MOD group.The results showed that SIRT1 inhibitor could block the effect of EA and weaken the effect of EA on the acetylation of NF-κB.This confirms that EA regulate the proportion of NF-κB acetylation in adipose tissue by activating SIRT1.(2)The co-expression results of SIRT1/Ac-NFκB showed that SIRT1 and Ac-NFκB were co-expressed in adipocytes,and the co-expression sites were mainly in the nucleus.According to the ratio of SIRT1/Ac-NFκB,the value in the MOD group was apparently lower than that in the NOR group(P<0.01).This results suggest that the content of SIRT1 in adipose tissue nucleus of insulin-resistant obese rats is on the low side,while the proportion of Ac-NFκB is on the high side.Compared with the MOD group,the ratio of the EA group was significantly increased(P<0.01).It can be seen that EA elevated the content of SIRT1 in the adipose tissue nucleus of insulin-resistant obese rats and reduce the content of Ac-NFκB.There was no difference between the SA group and the MOD group.Visibly,the non-acupoint acupuncture has no promotion on the content of SIRT1 and Ac-NFκB in the nucleus,which also confirms the effect of EA.Compared with the MOD group,the ratio of SIRT1/Ac-NFκB was significantly upgraded in the RSV group(P<0.01).This indicates that activation of SIRT1 can increase the relative expression of SIRT1 in adipose tissue nuclei and decrease the ratio of Ac-NFκB.There was no statistical difference in SIRT1/Ac-NFκB ratio between the EI and the MOD group,suggesting that the SIRT1 content in the adipose tissue nuclei of the EI group was low,while the ratio of Ac-NFκB was high.The results showed that SIRT1 inhibitor blocked the regulation of SIRT1 and Ac-NFκB by EA.The promotion in the ratio of the EA group was related to the activation of SIRT1 by EA.Since the results of the immunofluorescence double label are only semi-quantitative,the expression level of the above specific protein is still based on the protein quantification results of WB.4.EA reduces the expression of acetylated H3K9 in adipose tissue and the degree of acetylation of H3K9 in the NF-κB promoter region.(1)The protein expression result of Ac-H3K9: Compared with the NOR group,the expression of Ac-H3K9 in the adipose tissue nucleus of the MOD group was upgraded(P<0.05),suggesting that the acetylation degree of histone H3K9 increased in the adipose tissue nucleus of insulin-resistant obese rats.Compared with the MOD group,the expression of Ac-H3K9 in the EA group was obviously decreased(P<0.05).This indicates that the intervention of EA reduce the degree of acetylation of histone H3K9 in the nucleus of adipose tissue.There was no statistical difference between the SA group and the MOD group.It can be seen that non-acupoint acupuncture did not affect the degree of histone acetylation,which is a positive effect on EA.The expression of Ac-H3K9 in the nucleus of the RSV group was apparently decreased compared with the MOD group(P<0.05),which further confirmed that the activation of SIRT1 is closely related to the degree of acetylation of histone H3K9 in the adipose tissue nuclei of insulin resistant obese rats.There was no difference in the expression of Ac-H3K9 in the adipose tissue nuclei between the EI group and the MOD group.This indicates that SIRT1 inhibitors block the activation of SIRT1 by EA to a certain extent,resulting in an increase in the degree of acetylation of H3K9.(2)The results of SIRT1/Ac-H3K9 co-expression showed that SIRT1 and Ac-H3K9 were co-expressed in the nucleus of adipocytes,clearly seen in each group of 400 X microscopes.From the ratio of SIRT1/Ac-H3K9,the ratio of the MOD group was significantly lower than that of the NOR group(P<0.05),indicating that the ratio of SIRT1 in the adipose tissue nucleus of insulin-resistant obese rats was decreased,while the acetylation of H3K9 was elevated.After EA intervention,the ratio of SIRT1/Ac-H3K9 was obviously increased compared with the model group(P<0.01),suggesting that EA can upgrade the expression of SIRT1 in the adipose tissue nucleus of insulin-resistant obese rats and reduce the acetylation degree of H3K9.There was no apparent difference between the SA group and the MOD group.It can be seen that the non-acupoint acupuncture did not improve the degree of SIRT1 and the acetylation of H3K9 in the adipose tissue nucleus.Compared with the MOD group,the SIRT1/Ac-H3K9 ratio was significantly raised in the RSV group,which confirmed that the SIRT1 agonist activate SIRT1 and reduce the degree of acetylation of H3K9 in the nucleus.There was no statistical difference between the EI group and the MOD group,indicating that SIRT1 inhibitor blocked the effect of EA on SIRT1 to some extent,which lead to an increase in the degree of acetylation of H3K9 in the nucleus.(3)The degree of acetylation of H3K9 in NF-κB promoter region: Compared with the NOR group,the content of Ac-H3K9 in the promoter region of NF-κB gene in the MOD group was significantly increased(P<0.05).This indicates that the degree of acetylation of H3K9 in the NF-κB gene promoter region in the adipose tissue nuclei of insulin-resistant obese rats is upgraded.After EA intervention,the degree of acetylation of H3K9 in the NF-κB promoter region was obviously decreased(P<0.05).There was no statistical difference between the NOR group and the EA group.Visbily,the intervention of EA restore the acetylation degree of H3K9 in the promoter region of NF-κB gene in the adipose tissue nucleus of insulin-resistant obese rats to a normal level.Conclusions 1.EA can reduce the body weight and Lee’s index of insulin-resistant obese rats,and significantly lower the levels of CRP,IL-6 and TNF-α in peripheral blood,and improve the insulin resistance.2.EA can improve the infiltration of macrophages in adipose tissue of insulin-resistant obese rats.In addition,EA promote the transformation of M1 macrophages into M2 macrophages in adipose tissue of insulin-resistant obese rats,which may be related to the up-regulation of SIRT1 by EA.3.EA alters the expression of SIRT1 in adipose tissue of insulin-resistant obese rats.SIRT1 deacetylates of Ac-NFκB,thereby reducing the transcription of IL-6 and TNF-α in adipose tissue.4.EA can increase the expression of SIRT1 in adipose tissue and deacetylate of Ac-H3K9,thereby decreasing the degree of acetylation of histone H3K9 in the promoter region of NF-κB gene.This improves chromatin density and reduce the binding of NF-κB to promoter gene,and control the transcription of the downstream rlated genes.5.Resveratrol can effectively activate the expression of SIRT1 in adipose tissue of insulin-resistant obese rats.It deacetylates on acetylated NF-κB and histone H3K9,inhibits the transcription of NF-κB,and controls the expression of inflammatory factors in adipose tissue of rats,reduces the inflammatory state of obese rats and improves insulin sensitivity.Non-acupoint acupuncture treatment did not improve the inflammatory state and insulin sensitivity of insulin-resistant obese rats.SIRT1 inhibitor—sirtinol can partially block the activation of SIRT1 by EA,and reduces the anti-inflammatory effect of EA and improves insulin sensitivity to a certain extent.In summary,It is one of the mechanisms to improve insulin resistance in obesity via regulation the epigenetic modification of SIRT1/NF-κB signaling pathway by EA.This provides a theoretical basis for EA to prevent obesity and insulin resistance-related di SAses. |
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