Study of Early Growth Response Gene-1 DNA Enzyme on Preventing Stenosis and Occlusion of Autogenous Vein Graft:An Experiment with Rats

Objective To detect the inhibitory effect of early growth response gene-1 DNA enzyme(EDRz) on proliferation of vascular smooth muscle cell(VSMC) and intimal hyperplasia,and confirm the effect of gene therapy on stenosis and occlusion after vein transplantation. Methods EDRz was constructed,and autogenous vein graft model was established with Wistar rats,transplanting the right jugular vein to infra renal abdominal aorta by microsurgical technique.EDRz was transfected to the graft veins and the vein graft samples were harvested at hour 1,2,6,24 and on day 3,7,14,28,42 after grafting,10 Wistar rats were randomly selected in every time.The expression of EDRz in transfected vein graft was detected by fluorescent microscope.Egr-1 mRNA was measured by reverse transcription-PCR(RT-PCR) and in situ hybridization,respectively.The protein expression of Egr-1 was detected by Western blot and immunohistochemistry,respectively.HE stained vein grafts were observed under microscope. Results ① The results of EDRz transfected vein graft: At hour 1 after grafting,EDRz was mainly located in adventitia,tunica media,and partial endothelial cells of vein graft;At hour 2,6,and 24,EDRz was located in tunica media of vein graft;and on day 7,it was mainly located in intima of vein graft.There wasn't EDRz in vein grafts on day 14,28,and 42.② The results of expression of Egr-1 mRNA: Detection by RT-PCR: At hour 1 after transfecting,the expression of Egr-1 mRNA arrived at the peak,and declined at hour 2,6,and 24.The expression was tenuity on day 3.Egr-1 mRNA expression was not found on day 7,14,28,and 42.The expression of Egr-1 mRNA at hour 1 was significantly higher than that of the other time point(P0.01).The result of in situ hybridization was coincident with RT-PCR.③ The results of expression of Egr-1 protein: The result of Western blot: There was no expression of Egr-1 protein in normal veins.At hour 2 after grafting,expression of Egr-1 protein was found,and declined at hour 6,24,and on day 3.There was no expression of Egr-1 protein at hour 1,and on day 7,14,28,and 42.The expression of Egr-1 protein at hour 2 was significantly higher than that of the other time point(P0.01).The result of immunohistochemistry was coincident with Western blot.④The degree of VSMC hyperplasia and intimal thickness were lighter in EDRz transfected vein grafts than that in not-transfected vein grafts contemporarily. Conclusions EDRz could reduce the expression of Egr-1 in autogenous vein graft,and could effectively restrain VSMC proliferation and intimal hyperplasia,and prevent vascular stenosis and occlusion after vein grafting.