Effects Of EPO On Serum Inflammatory Cytokines IL-10?TGF-??IL-2 And IL-8 Levels After Cardiac Transplantation In SD Rats

Objective:We previously found a phenomenon that a huge number of inflammatory cells infiltrated into the myocardium after cardiac transplantation in homologous SD rats,resulting in serious impact on the cardiac function after heart transplantation.Therefore,in the present study we intend to further detect the effect of EPO on the levels of serum inflammatory cytokines IL-10,TGF-?,IL-2 and IL-8 and inflammatory infiltration of myocardium in homologous SD rats after heart transplantation,so as to uncover its cardio-protection function through reducing inflammatory infiltration after heart transplantation.Methods:1.Healthy male SD rats(n=90)are divided into sham Operation Group(Group A,n=18),Simple Heart Transplantation Group(Group B,n=36,donor 18,receptor 18),Heart transplant +EPO Group(Group C,n=36,donor 18,receptor 18)according to the random digital table.The sham operation group only undergo laparotomy,the heart transplant group undergo abdominal heterotopic heart transplantation,cardiac transplant+EPO Group undergo heterotopic heart transplantation and EPO intervention,and the bloods and hearts in each group are collected at 1d and 5d post-operation.The blood samples are used to detect the levels of myocardial enzymes,IL-10,TGF-?,IL-2,and IL-8,and the myocardial tissue specimens are used to observe the infiltration and distribution of inflammatory cells through HE staining.2.Animal Model : donor recumbent in constant operation bed after ether anesthesia,are intravenously injected with heparin saline 0.5 ml(20 iu/ml)to make the whole body heparinization through vena cava after thoracotomy.Dissociate and snip the thoracic aorta and pulmonary artery,then clip the rest blood vessels,and the heart preserves in the ice cold salt water.Receptor is cut open in the abdominal midline after anesthesia,fully exposing abdominal aorta and inferior vena cava for at least 4 mm,and blocking microvascular clip after all the cut out appropriate size anastomotic.Remove the donor heart,the aorta and recipient abdominal aorta,pulmonary artery and inferior vena cava respectively match in a row after the completion of the slowly restoring blood supply.The transplanted heart beat recovery in several minutes.Then close abdomen and supply the cage with water and food,and keep the surgery rats in constant temperature preservation until awakening.Results:1.Establishment of rats transplantation model:(1)Two cases die of intestinal obstruction,and one case dies of anesthesia accident,and there is no significant difference in the survival rate of each group at each point(P>0.05).(2)Heart beat: There is no significant changes in the beating strength at 1d in group B and C.The heart beats in Group B decreased markedly comparing with group C at day 5;(3)Myocardial enzymes: Compared with group A,the activities of serum creatine kinase and lactate dehydrogenase phosphate increased significantly in group B at day 1 and 5 after transplantation(P< 0.05).Compared with group C,the activities of serum creatine kinase and lactate dehydrogenase phosphate increased significantly in group B at day 1 and 5 after transplantation(P < 0.05).2.HE staining: In Group A,group B,and Group C,the structure of the myocardial cells are intact at day 1,with regular myocardial fiber arrangement but a bit of inflammatory cell infiltration,without local bleeding and necrosis.In group B at day 1 after transplantation,myocardial cells are swelling and badly damaged,with irregular muscle fiber arrangement and increasing inflammatory cell infiltration.In Group C,there are a small number of muscle fibers rearranged and inflammatory cell infiltration at day 1 after operation.In group B at day 5 after transplantation,both of inflammatory cell infiltration and myocardial cell necrosis increase,with myocardial fiber structure disorder.However,in group B at day 5 after transplantation,there is only a bit of increasing of inflammatory cell infiltration,and the structure of myocardium is still regular.3.The levels of inflammatory cytokines in IL-10,TGF-?,IL-2 and IL-8: At day 1 and 5after heart transplantation,compared with group C,the serum IL-2 and IL-8 levels increased significantly in group B;compared with group C,the levels of IL-10 and TGF-?significantly increased in Group C(P<0.05).Conclusion:EPO play the role of cardio-protection on heart transplantation via affecting the levels of serum cytokines that are IL-2,IL-8,IL-10 and TGF-?.