| Objective:To explore the effect of acupuncture and postconditioning on the expression of opioid receptor and RISK pathway in heart tissue of rats with myocardial ischemia-reperfusion injury,and to provide a new experimental basis for the study on the mechanism of post-treatment of acupuncture treatment of myocardial ischemia-reperfusion injury.Methods:140 SD rats were randomly divided into blank group,model group,sham acupuncture group,acupuncture group before reperfusion,acupuncture group after reperfusion,acupuncture group before and after reperfusion,and ?-OPR antagonist group total 7 Groups,20 in each group.In addition to the blank group,the model of myocardial ischemia-reperfusion injury was established by ligating left anterior descending coronary artery for 40 min and reperfusion for 120 min.Electrocardiograph was used to detect electrocardiogram.Tunel technique was used to detect cardiomyocyte apoptotic rate.Myocardial infarct size was detected by TTC staining.Myocardial oxygen consumption was calculated by heart rate and left ventricular arterial pressure.Real-time fluorescence quantitative PCR was used to detect rat heart.The expression of ?-OPR mRNA was detected by Western blot.The expression of ?-OPR protein was detected by Western blot.The expression of Akt,Erk,JNK,and p38 was detected by Luminex liquid chip technique.All data were statistically analyzed with statistical software SPSS 24.0 and the results were obtained.Result:1.Heart function index detection1.1 ST-segment changes: ST-segment elevation in the model group,sham acupuncture group,both acupuncture group and ?-OPR antagonist group before and after reperfusion was significantly higher than that of the blank group(P<0.01);Compared with before and after reperfusion,both the acupuncture group and the ?-OPR antagonist group significantly decreased the ST segment elevation(P<0.01).The sham acupuncture group showed a slight decrease,but the difference was not significant(P>0.05);before and after reperfusion Compared with the acupuncture group,ST elevation in the ?-OPR antagonist group and the sham acupuncture group was significantly higher(P<0.01).1.2 Average apoptotic rate of cardiomyocytes: Compared with the model group,the average apoptotic rate of myocardial cells in the acupuncture group and ?-OPR antagonist group was significantly decreased before and after reperfusion(P<0.01),and the sham acupuncture group was significantly decreased(P<0.01).<0.05);Compared with the acupuncture group before and after reperfusion,the average apoptotic rate of myocardial cells in the ?-OPR antagonist group and the sham acupuncture group was significantly increased(P<0.01).1.3 Myocardial Infarct Size: Compared with the model group,the myocardial infarct size in the acupuncture group and ?-OPR antagonist group was significantly reduced before and after reperfusion(P<0.01),and the sham acupuncture group was slightly reduced,but the difference was not significant(P> 0.05);Compared with the acupuncture group before and after reperfusion,the myocardial infarct size in the ?-OPR antagonist group and the sham acupuncture group increased significantly(P<0.01).1.4 Myocardial Oxygen Consumption: Compared with the blank group,myocardial oxygen consumption of the model group,sham acupuncture group,both the acupuncture group before and after reperfusion,?-OPR antagonist group was significantly reduced(P<0.01);The oxygen consumption of myocardium in the acupuncture group,?-OPR antagonist group,and sham acupuncture group was significantly increased before and after reperfusion(P<0.01).Compared with the acupuncture group before and after reperfusion,the ?-OPR antagonist group was Myocardial oxygen consumption was slightly increased,but the difference was not significant(P>0.05),and the sham acupuncture group was significantly lower(P<0.01).2.? Opioid receptor expression levels2.1 ?-OPR mRNA expression: compared with blank group,?-OPR mRNA expression in model group,sham acupuncture group,reperfusion before acupuncture group,acupuncture group after reperfusion,acupuncture group before and after reperfusion The amount was significantly increased(P<0.01);compared with the model group,the expression of ?-OPR mRNA was significantly increased in the acupuncture group before reperfusion,the acupuncture group after reperfusion,and the acupuncture group before and after reperfusion(P<0.01).),sham acupuncture group slightly reduced,but the difference was not significant(P> 0.05);compared with reperfusion before and after acupuncture group,reperfusion before acupuncture group,reperfusion after acupuncture group,sham acupuncture group The expression of ?-OPR mRNA was significantly decreased(P<0.01).2.2 ?-OPR protein expression: compared with blank group,model group,sham acupuncture group,acupuncture group before reperfusion,acupuncture group after reperfusion,?-OPR protein expression in acupuncture group before and after reperfusion The amount increased significantly(P<0.01).Compared with the model group,the expression of ?-OPR protein in the acupuncture group before reperfusion,the acupuncture group after reperfusion,and the acupuncture group before and after reperfusion significantly increased(P<0.01).),sham acupuncture group increased slightly,but the difference was not significant(P> 0.05);compared with reperfusion before and after the acupuncture group,reperfusion before acupuncture group,reperfusion after acupuncture group,sham acupuncture group The expression of ?-OPR protein was significantly reduced(P<0.01).3.Detection of RISK conduction pathways3.1 Akt(Ser473): Compared with the blank group,Akt(Ser473)expression in the model group,acupuncture group,?-OPR antagonist group,and sham acupuncture group before and after reperfusion significantly decreased(P<0.01);Compared with before and after reperfusion,the expression of Akt(Ser473)in the acupuncture group and ?-OPR antagonist group was significantly increased(P<0.01),but the sham acupuncture group increased slightly,but the difference was not significant(P>0.05);Compared with the acupuncture group before and after perfusion,the expression of Akt(Ser473)in the ?-OPR antagonist group and the sham acupuncture group was significantly decreased(P<0.01).3.2 Erk1/2(Thr185/Tyr187): Compared with the blank group,Erk1/2 expression in the acupuncture group,?-OPR antagonist group,and sham acupuncture group before and after reperfusion was significantly decreased(P<0.01);Compared with the model group,the expression of Erk1/2 in the acupuncture group and ?-OPR antagonist group was significantly increased before and after reperfusion(P<0.01).The sham acupuncture group had a slight decrease,but the difference was not significant(P>0.05).Compared with the acupuncture group before and after reperfusion,the expression of Erk1/2 in the ?-OPR antagonist group was significantly lower(P<0.05),and the sham acupuncture group was significantly lower(P<0.01).3.3 JNK(Thr183/Tyr185): compared with the blank group,JNK expression levels in the model group,acupuncture group before,after reperfusion,?-OPR antagonist group,sham acupuncture group decreased significantly(P <0.01);The expression of JNK in the acupuncture group and ?-OPR antagonist group was significantly increased before and after reperfusion(P<0.01).The sham acupuncture group had a slight increase,but the difference was not significant(P>0.05);and the acupuncture before and after reperfusion Compared with the group,the expression of JNK in the ?-OPR antagonist group was significantly lower(P<0.05),and the sham acupuncture group was significantly lower(P<0.01).3.4 p38(Thr180/Tyr182): compared with the blank group,the expression of P38 in the model group,the acupuncture group,?-OPR antagonist group,and sham acupuncture group before and after reperfusion significantly decreased(P<0.01);The expression of P38 in the acupuncture group and ?-OPR antagonist group was significantly increased before and after reperfusion(P<0.01).The sham acupuncture group had a slight increase,but the difference was not significant(P>0.05);and the acupuncture before and after reperfusion Compared with the group,the expression of P38 in the ?-OPR antagonist group and the sham acupuncture group was significantly decreased(P<0.01).Conclusion:1.Post-treatment by acupuncture can effectively reduce the rate of cardiomyocyte apoptosis,decrease myocardial infarct size,reduce myocardial oxygen consumption,and reduce the elevation of ST segment in electrocardiogram.It can protect the myocardium from ischemia-reperfusion injury in rats.2.Acupuncture can increase the content and activity of ?-OPR,and the effect of acupuncture treatment before and after ischemia is significantly better than that of acupuncture before ischemia and post-ischemic treatment;3.?-OPR antagonists can significantly reverse the effect of acupuncture treatment,suggesting that the effect of acupuncture on improving myocardial ischemia-reperfusion injury may depend on the activation of ?-OPR receptor-mediated signaling pathway.4.Acupuncture treatment can significantly up-regulate the expression of ?-OPR,increase the phosphorylation of Akt,Erk,JNK and p38 in cardiac tissue,activate the RISK pathway,and thus reduce the damage caused by myocardial ischemia and reperfusion to protect the heart. |
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