OBJECTIVE:To investigate the effect of electroacupuncture(EA)on regulatory functions of one specific exosomal circ RNA of Enhancer of Zeste Homolog(Circ EZH)and its potential mechanisms of action in type 2 diabetes(T2...OBJECTIVE:To investigate the effect of electroacupuncture(EA)on regulatory functions of one specific exosomal circ RNA of Enhancer of Zeste Homolog(Circ EZH)and its potential mechanisms of action in type 2 diabetes(T2DM).METHODS:Mice were fed a high-fat diet(HFD)and intraperitoneally injected with streptozotocin to create the T2DM model and then were used for two experiments involving the following groups:experiment 1(control group,T2DM group,T2DM+EA group,10 mice per group)and experiment 2[control group,T2DM group,T2DM+Circ EZH1-si RNA(20 nmol/20 g)group,10 mice per group].Exosomal size,distribution,and morphology were evaluated via transmission electron microscopy and nanoparticle tracking analysis.The expression of Circ EZH1 and Circ EZH2 in exosomes was assessed by quantitative real-time polymerase chain reaction.Insulin expression was assessed by enzyme-linked immunosorbent assay,immunofluorescence,and western blotting.The effects of exosomal Circ EZH1 knockout and overexpression in Min6 cells were assessed by cell counting kit-8 and flow cytometry.Meanwhile,the effect of Circ EZH1 knockout on insulin sensitivity was assessed by glucose tolerance test(GTT)and insulin tolerance test(ITT)in vivo.RESULTS:EA treatment significantly reduced the serum insulin level and cell apoptosis in pancreatic tissue in a T2DM model.EA treatment markedly upregulated Circ EZH1 expression in the exosomes of T2DM mice.Further study showed that Circ EZH1 overexpression resulted in increased Min6 cell viability and decreased Min6 cell apoptosis when compared with the levels in an overexpression control group.In Min6 cells with Circ EZH1 knockout,the opposite trends were identified.Circ EZH1-knockout Min6 cells also showed reduced insulin expression.In vivo,Circ EZH1-knockout T2DM mice displayed damaged insulin sensitivity,which was demonstrated by elevated levels of fasting blood glucose and decreased glucose tolerance in the GTT and insulin sensitivity in the ITT.CONCLUSIONS:EA can affect Circ EZH1 expression specifically in the exosomes inβcells in the pancreatic islets to improve T2DM.Exosomal Circ EZH1 is a potential therapeutic candidate to treat T2DM.展开更多
基金Science and Technology Committee of Songjiang District,Shanghai(2024SJKJGG001)project:Mechanistic Study on the Therapeutic Effect of Electroacupuncture in Diabetic Osteoporosis via Exosomal circular RNA of Enhancer of Zeste Homolog 1-Regulated miR-128/Transient Receptor Potential Vanilloid 1 Axis-Mediated Ferroptosis in Osteoclasts+9 种基金the Shanghai Municipal Health Bureau[grant number:2020LP010]Project:a Randomized Controlled Study of Traditional Chinese Acupuncture Combined with Rehabilitation Training for the Treatment of Intensive Care UnitAcquired Weaknessthe Science and Technology Commission of Shanghai Municipality(grant number:20511101204)Project:Traditional Chinese Medicine Data Collection and Governance for Early Screening and Stratified Diagnosis and Treatment of Pancreatic Cancerthe Shanghai University of Traditional Chinese Medicine(grant number:2021LK100)Project:the Role of Sestrin 2/mechanistic Target of Rapamycin-Mediated Autophagy in Age-Related Skeletal Muscle Atrophy and the Effects of Acupuncturethe Shanghai General Hospital(grant number:202220)Project:Research on the Construction of Talent Evaluation System by the Southern Outpatient Party BranchShanghai Municipal Commission of Health and Family Planning[grant number:ZY(2021-2023)-0208]Project:Special Program of the Integrated Chinese and Western Medicine Innovation Research Institute。
文摘OBJECTIVE:To investigate the effect of electroacupuncture(EA)on regulatory functions of one specific exosomal circ RNA of Enhancer of Zeste Homolog(Circ EZH)and its potential mechanisms of action in type 2 diabetes(T2DM).METHODS:Mice were fed a high-fat diet(HFD)and intraperitoneally injected with streptozotocin to create the T2DM model and then were used for two experiments involving the following groups:experiment 1(control group,T2DM group,T2DM+EA group,10 mice per group)and experiment 2[control group,T2DM group,T2DM+Circ EZH1-si RNA(20 nmol/20 g)group,10 mice per group].Exosomal size,distribution,and morphology were evaluated via transmission electron microscopy and nanoparticle tracking analysis.The expression of Circ EZH1 and Circ EZH2 in exosomes was assessed by quantitative real-time polymerase chain reaction.Insulin expression was assessed by enzyme-linked immunosorbent assay,immunofluorescence,and western blotting.The effects of exosomal Circ EZH1 knockout and overexpression in Min6 cells were assessed by cell counting kit-8 and flow cytometry.Meanwhile,the effect of Circ EZH1 knockout on insulin sensitivity was assessed by glucose tolerance test(GTT)and insulin tolerance test(ITT)in vivo.RESULTS:EA treatment significantly reduced the serum insulin level and cell apoptosis in pancreatic tissue in a T2DM model.EA treatment markedly upregulated Circ EZH1 expression in the exosomes of T2DM mice.Further study showed that Circ EZH1 overexpression resulted in increased Min6 cell viability and decreased Min6 cell apoptosis when compared with the levels in an overexpression control group.In Min6 cells with Circ EZH1 knockout,the opposite trends were identified.Circ EZH1-knockout Min6 cells also showed reduced insulin expression.In vivo,Circ EZH1-knockout T2DM mice displayed damaged insulin sensitivity,which was demonstrated by elevated levels of fasting blood glucose and decreased glucose tolerance in the GTT and insulin sensitivity in the ITT.CONCLUSIONS:EA can affect Circ EZH1 expression specifically in the exosomes inβcells in the pancreatic islets to improve T2DM.Exosomal Circ EZH1 is a potential therapeutic candidate to treat T2DM.
