p53基因转移至移植心脏的安全性

Security for adenovirus-mediated p53 gene transfer to the donor heart

背景:课题组前期实验表明野生型p53基因具有抑制移植心脏冠状动脉内膜增厚的作用。目的:研究腺病毒介导的野生型p53基因转移至移植心脏的安全性。方法:以Wistar大鼠为供体,SD大鼠为受体建立大鼠腹腔异位心脏移植模型,在取出供心后,经供心冠状动脉分别注射携带野生型p53基因的重组腺病毒液、携带β-半乳糖酐酶基因的重组腺病毒液和生理盐水800μL,4℃静置30min后进行心脏移植。结果与结论:移植后5d,重组腺病毒液组的供心冠状动脉组织可见野生型P53蛋白的表达。移植后28d,未见受体大鼠血液生化学指标异常和重要脏器的病理性改变,RT-PCR扩增未见腺病毒E1A区产物。说明在心脏移植中,将腺病毒介导的野生型p53基因转移至移植心脏是安全的。

BACKGROUND： Wild-type p53 gene transfer to the donor heart can greatly inhibit graft coronary artery intima hyperplasia and lumen narrowness. OBJECTIVE： To study the security of adenoviral-mediated wild-type p53 gene transfer to the donor heart after heart transplantation. METHODS： Rat models of heterotopic （abdomen） heart transplantation were developed. Wistar rats served as donors and SD rats as recipients. After donor hearts were removed, 800 ？L adenoviral vector encoding the wild-type p53 gene （Ad-p53 group）, adenoviral vector encoding the β-galactosidase gene （LacZ） （Ad-LacZ group） or saline （control group） were infused into the donor heart respectively before transplantation. The donor heart was stored in the 4 ℃ saline for 30 minutes before heart transplantation. At 5 days after operation, P53 protein expressions in coronary artery of donor hearts were tested by western blot analysis. At 28 days after transplantation, the serum specimen was collected for the biochemical indicators, and the major organs of the recipients were tested by the histopathological analysis and the reverse transcription polymerase chain reaction of the adenoviral E1A sequences. RESULTS AND CONCLUSION： The expression of P53 protein was found in donor hearts in Ad-p53 group at 5 days after operation, and no expression in Ad-LacZ group and control group. At 28 days after operation, rat serum biochemistry values in three groups was normal, the major organs of the recipients were not affected seriously, no virus spread to other organs in this experimental protocol. The results confirmed that the ex vivo adenoviral-mediated gene transfer to the donor heart via the coronary artery during the heart transplantation is safe.