The Research Of Endogenous Opioid Peptides Mechanism In The Protective Effect Of Electroacupuncture On Acute Myocardial Ischemia Rabbits

The Research of Endogenous Opioid Peptides Mechanismin the Protective Effect of Electroacupuncture onAcute Myocardial Ischemia RabbitsSpeciality:Acupuncture and MassageAuthor:You XinghongTutor:Wang Hua ProfessorObjectiveTo explore both the effect and the mechanisms of Shuangguyitongtherapy of acupuncture in the rabbits with acute myocardialischemia(AMI);Yoresearch theroleofendogenousopioidpeptidesof anti-AMI effect of Shuangguyitong therapy of acupuncture on theAMI rabbits.Methods80 rabbits were evenly and randomly divided into 4 groups:blankcontrol group(blank,non-model and non-needling,n=10);modelgroup(model and non-needling,n=10);electric needle group(modeland Shuangguyitong therapy of acupuncture electric needlingGuanyuan,Zusanli and Neiguan acupoint,n=10).naloxone group(microinjection naloxone into rostral ventrolateral medulla,modeland Shuangguyitong therapy of acupuncture electric needlingGuanyuan,Zusanli and Neiguan acupoint,n=10).Afteranesthesia(20% urethane,5ml/kg,i.p.)andmicroinjection naloxoneor sodium chloride into rostral ventrolateral medulla,AMI rabbitmodels were induced by ligating left ventricular branch of coronaryartery,treated by Shuangguyitong therapy of acupuncturestimulated with electric needling Guanyuan,Zusanli and Neiguan acupoint.To record theⅡleadECGand meanarterialpressureat5 min,15 min,30 min,45 min,60 min after acute myocardial ischemiain rabbits.To observe the change of AMI and myocardialultrastructures of rabbit' sischemicmyocardial cell.Contents ofDynorphin(Dyn A_1-13),methionine enkephalin(M-ENK)and leuthineenkephalin(L-ENK)in brain tissue and plasma were determined byradioimmunoassay.Investigate the distribution of leuthineenkephalin immunoreactive fibers in the medulla byimmunohistochemical method.To explore the preproenkephalin mRNAexpression of medulla by the method of in situ hybridizationhistochemistry after electroacupuncture.Results1.EGG:Compared with blank control group,the changes of T waveand ST segment of ECG differd significantly in model group andelectric needle group after ischemia(P＜0.01),no significant dataof model group showed that there was the difference in 60 minutes.Shuangguyitong therapy of acupuncture electric needling Guanyuan,Zusanli and Neiguan acupoint could markedly promote recovery of EGGafter ischemia,there was significant difference than model group(P＜0.05).The protective effect can be weakened by microinjectionwithNaloxoneintorVLM(P＜0.05),but can' tbecompletelyblocked.2.Meanarterial pressure:Compared with blank control group,themean arterial pressure depressed significantly in model group andelectric needle group after ischemia(P＜0.01),Shuangguyitongtherapy of acupuncture electric needling Guanyuan,Zusanli andNeiguan acupoint could markedly promote recovery of mean arterialpressure after ischemia,there was significant difference thanmodel group(P＜0.05).The effect can be weakened by microinjection with Naloxone into rVLM(P＜0.05).3.Concentration of endogenousopioidpeptides inplasma:Comparedwith blank control group,Concentration of Dyn A_1-13),M-ENK and L-ENKincreased markedly in plasma in model group(P＜0.01)Theconcentration of Dyn A_1-13),M-ENK and L-ENK in plasma afterShuangguyitong therapy of acupuncture electric needling Guanyuan,Zusanli and Neiguan acupoint increased significantly compared tothe model group(P＜0.01)4.Concentration of endogenous opioidpeptides in brain:Comparedwith blank control group,Concentration of Dyn A_1-13),M-ENK and L-ENKincreased markedly in medulla and hypothalamus in model group(P＜0.01).The concentration of Dyn A_1-13),M-ENK and L-ENK in plasmaafter Shuangguyitong therapy of acupuncture electric needlingGuanyuan,Zusanli and Neiguan acupoint increased significantlycompared to the model group(P＜0.01).5.Myocardial tissue injury:Compared with blank control group,tthetissue injury of myocardium examined with optical microscope inmodel group were obvious Microinjection naloxone into rVLM hasno influence to normal rabbit' s myocardial tissue.Shuangguyitongtherapy of electroacupuncture could obviously reduce the degree ofmyocardium ischemic injury of myocardial ultrastructures inrabbits.The effect can be weakened by microinjection with Naloxonein rVLM.6.Myocardial ultrastructures:Microinjectionnaloxone into rVLMhas no influence to normal rabbit' s myocardial ultrastructures.Electroacupuncture has significant protective effect to acutemyocardium ischemic in rabbits,results indicated thatShuangguyitong therapy of electroacupuncture could obviously reduce the degree of myocardium ischemic injury of myocardialultrastructures in rabbits.In the electroacupuncture group,lessinjury of myocardial tissue,normal run of myocardial fiber,a smallamount mitochondrial vacuolation and no obvious dissolution andnecrosis of myocardium were found.The protective effect can beweakened by microinjection with Naloxone in rVLM.7.L-ENK immunoreactive neurons in the medulla tissue:L-ENKimmunoreactive neurons were mainly distributed in the nucleitractus solitarii(NTS),ventrolateral medulla(VLM)and thereticular formation betweenNTS andVLM,forming an arc band zonefrom dorsomedial part to ventrolateral part of medulla.Comparedwith blank control group,these structures ofL-ENK immunoreactivebodies and dibers were increased markedly in medulla in model group(P＜0.01).After Shuangguyitong therapy of acupuncture electricneedlingGuanyuan,ZusanliandNeiguanacupoint,the expression ofL-ENK immunoreactive bodies were increased significantly comparedto the model group(P＜0.01).8.PPEmRNA containing neurons in the medulla tissue:PPEmRNAcontaining neurons were also densely seen in the medullastructures.Compared with blank control group,these structures ofPPEmRNA containing neurons were increased markedly in medulla inmodel group(P＜0.01).After Shuangguyitong therapy of acupunctureelectric needling Guanyuan,Zusanli and Neiguan acupoint,theexpression of PPEmRNA containing neurons were increasedsignificantly compared to the model group(P＜0.01)Comclusions1.Shuangguyitong therapy of acupuncture electric needlingGuanyuan,Zusanli and Neiguan acupoint could markedly promote recovery of the T wave and ST segment of ECG and mean arterialpressure after myocardial ischemia and could obviously reduce thedegree of myocardium ischemic injury of myocardial tissue inrabbits.2.The protective effect of Shuangguyitong therapy ofelectroacupuncture in ameliorating acute myocardium ischemic inrabbits was mediated by endogenous opioid peptides in plasma.3.Shuangguyitong therapy of acupuncture could protect ischemicmyocardium cell after myocardial ischemia.The effect ofameliorating acute myocardium ischemic in rabbits was mediated byendogenous opioid peptides in brain.4.The effect of Shuangguyitong therapy of electroacupuncture inameliorating acute myocardium ischemic in rabbits was mediated byopioid receptors in rVLM.5.Shuangguyitong therapy of acupuncture electric needlingGuanyuan,Zusanli and Neiguan acupoint can significantly improvethe expression of L-ENK immunoreactive neurons and PPEmRNAcontaining neurons in the VLM.The protective effect ofelectroacupuncture was mediated by opioid receptors in VLM.