缺氧诱导因子-1α与同源盒基因共转染在移植静脉重塑态的表达

Expression of hyopxia inducible factor-la and gax genes on the intimal hyperplasia for venous autografts in rat

目的探讨缺氧诱导因子（HIF-1α）和同源盒基因（gax）共转染在移植静脉组织中的表达状态。方法Wistar大鼠16只随机分为实验组和对照组，每组8只，均行自体静脉移植术，实验组移植静脉行缺氧诱导因子（HIF-1α）和同源盒基因（gax）共转染，对照组则未行基因转染，于术后14d取出移植的静脉，分别采用逆转录一聚合酶链反应（RT—PCR）、Western印迹（Western blotting）及透射电镜等技术检测，分析移植静脉中HIF-1α和gax基因的核酸及蛋白表达水平，血管内皮细胞（VEC）及血管平滑肌细胞（VSMC）表型的变化。结果HIF-1α的mRNA表达水平实验组为（93．1±22．9）bP，对照组为（42．7±15．9）bP，P〈0．01；gax基因的mRNA表达水平组（106．9±38．7）bP，对照组（50．7±25．8）bP，P〈0．01；HIF-1α的蛋白表达水平实验组（10．86±2．76）kD，对照组（4．52±1．90）kD，P〈0．01；gax基因的蛋白表达水平实验组（13．65±3．35）kD，对照组（5．40±2．26）kD，P〈0．01；VSMC合成表型实验组（21．3±5．4）％，对照组（41．2±8．6）％，P〈0．05。结论HIF-1α和gax基因联合转染可成功导入移植静脉组织并产生生物学效应。

Objective To explore the expression of （hyopxia inducible factor-a, HIF-1α） and gax genes on the intimal hyperplasia for venous autografts in rat. Methods Sixteen Wistar rats were randomLy divided into into 2 groups, a control group and an experimental group, 8 rats in each group. A rat model of venous autografts was established by transplanting the right external jugulars vein into right common carotid artery in 16 rats. The transplanted vein in the experimental group was immersed in solution with recombinant adeno-associated virus（rAAV） and HIF-1α by incubation for 45 minutes at 0℃-4℃ before anastomosis and coated using glue gel in which rAAV-gax added just after anastomosis, the control ones were immersed only in the physiological solution in which rAAV solution without HIF-1α gene added, nothing to be coated except for glue gel only. The vein grafts were taken at 14 days after operation, RT-PCR technique, Western blotting and electric microscopy were used to detect the expression of HIF-1α mRNA and protein, gax mRNA and protein, phenotypic switch of VSMC. Results The expression of HIF-1α mRNA, the experimental group （93.1 ± 22.9） bP, the con trol group（42.7 ± 15.9） bP,P 〈 0. 01 ; the exprossion of gax mRNA, the experimental group （ 106.9 ± 38.7） bP, the contrd group （50.7 ± 25.8 ） bP, P 〈 0.01 ; the exprossion of HIF-1α protein, the experimental group （ 10.86 ± 2.76） kD, the control group （4.52 ± 1.90） kD, P 〈 0.01 ; the expression of gax protein, the experimental group （ 13.65 ± 3.35 ） kD, the control group （5.40 -± 2.26） kD, P 〈 0.01 ; VSMC proliferation, the experinental group （21.3 ± 5.4）%, the control group （41.2 ± 8.6）%, P 〈 0.05. Conclusion The pres- ent study demonstrated that HIF-1α and gax genes transfection could inhibit proliferation of VSMC and their phenotypic conversion from contractile phenotype to synthetic phenotype in venous autografts.