CAJ | 학술논문

目的观察转化生长因子-β1(TGF.β1)转基因对大鼠移植心脏缺血-再灌注损伤的影响.方法利用Cuff技术建立心脏移植模型,转基冈组供心离体灌注携带mTGF-β1基因腺病毒颗粒;空白对照组灌注停跳液;空载体组灌注空白载体腺病毒颗粒.观察移植物超微结构变化;RT-PCR检测心肌组织mTGF-β1mRNA的转录强度;测定心肌组织中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量、髓过氧化物酶(MPO)活性.结果转基因组心肌组织有外源性mTGF-β1基凶及蛋白表达,心肌细胞凋亡显著减少、SOD活性升高、MDA含量和MPO活性下降.结论 TGF-β1基因能减轻移植心脏缺血.再灌注损伤.
목적 관찰전화생장인자-β1(TGF.β1)전기인대대서이식심장결혈-재관주손상적영향.방법 이용Cuff기술건립심장이식모형,전기강조공심리체관주휴대mTGF-β1기인선병독과립;공백대조조관주정도액;공재체조관주공백재체선병독과립.관찰이식물초미결구변화;RT-PCR검측심기조직mTGF-β1mRNA적전록강도;측정심기조직중초양화물기화매(SOD)활성、병이철(MDA)함량、수과양화물매(MPO)활성.결과 전기인조심기조직유외원성mTGF-β1기흉급단백표체,심기세포조망현저감소、SOD활성승고、MDA함량화MPO활성하강.결론 TGF-β1기인능감경이식심장결혈.재관주손상.
Objective Isohemia-reperfusion injury oecurred during heart transplantation may result in failure of grafts and the death of receivers perioperatively. Over expression of TGF-β1 in the myocardium therapeutically was shown to be help-ful in limiting the reperfusion injury to the grafts. The study was designed to investigate the role of Ad. mTGF-β1 gene transfec-tion during ischemia-reperfusion injury in vitro after heart transplantation in rats and the possible mechanisms. Methods The model of heterotopic cardiac transplantation was established by Heron's technique with cuff vessel anastomosis. Animals were divided into 3 groups: in group A ( n =6, control group), the donor hearts were perfusod with 6 ml of Stanford University cardio-plegic solution via coronary arteries at 4℃ for about 40 minutes; in group B ( n =6, vector alone group), the donor hearts were perfused with 6 ml of Stanford University cardioplegic solution containing 5 × 10~9 plaque-forming units( pfu)/gram of the vec-tor, and in group C (study group), the donor hearts were perfused with the solution containing 5 × 10~9 pfu/gram vector with mTGF-β1. The donor hearts were observed with an electro-microscope. The expression of mTGF-β1 in the grafts was identified with immunohistochemical staining. Gene products expressed in tissues were quantified by one step RT-PCR. Activities of SOD ,MDA ,MPO in the grafts were measured. Results At 8 hours after transplantation, mTGF-β1 and its expression were de-tected by means of RT-PCR and immunohistochemical staining in the rats of group C. Expression scores of foreign gene were significantly higher in groups A and B. The apoptotic index of the myocardial cells in group C was lower than those in groups A and B. The activity of SOD was higher in group C than those in groups A and B, though the activities of MDA and MPO were decreased. Conclusion The study demonstrated that gent transfer in vitro via coronary artery was effective. Ad. mTGF-β1 gene transfection in vitro ameliorates the myocardial ischemia-reperfusion injury in rats, for which heart transplantation was per formed, increases the activity of SOD, and decreases the activities of MDA, MPO.