【摘要】 目的 :研究经冠状动脉 (冠脉 )内导入腺病毒载体介导的重组 β gal基因后心肌的转染效率 ,以证明冠脉内导入途径是重组基因导入心脏的可行途径。方法 :将 1.5 3× 10 12 pfu重组 β gal腺病毒经冠脉造影导管由左冠脉分别导入到 7头雄性Yorkshire猪心脏中 ,于术后 1、2、4及 8周处死动物 ,取左右冠脉、3～ 5mm厚的不同部位的心肌 ,经 1.2 5 %戊二醛固定 2 0min ,用PBS洗涤 3次 ,在室温下用X gal染色液染色 16～ 2 4h ,脱水、石蜡包埋、切片及苏木精 伊红复染 ,光镜下计算出表达 β gal的心肌细胞百分数即为转染效率。 结果 :经冠脉内导入重组基因后 ,于术后 1周即有β gal表达 ,2周达高峰 ,心肌细胞的转染效率高达4 5 .5 7% ,左冠脉壁全层、心肌间质中均有β gal表达 ,4周 β gal表达下降 ,8周 β gal恢复至基线水平。结论 :经冠脉内导入重组腺病毒 ,心肌细胞及冠脉壁重组基因表达效率高 ,说明冠脉内导入重组目的基因可能是冠心病基因治疗中可行有效的方法更多还原

【Abstract】 Objective:To demonstrate the efficiency of trans catheter intracoronary infusion of recombinant adenovirus coding for β galactosidase gene. Methods:Left coronary artery was infused with 1.53 ×10 12 pfu recombinant adenovirus vector coding for β galactosidase gene via an intracoronary catheter in 7 male Yorkshire porcines. The porcines were rilled at one week, two weeks, four weeks and eight weeks after intracoronary infusion, and their hearts were harvested. β galactosidase gene expression was examined by X gal staining histochemically in these heart tissues. The efficiency of β galactosidase gene expression was estimated by determining the fraction of cells expressing β galactosidase gene in 10 high power microscopial fields.Results:β galactosidase gene expression was detected in both the myocardium and coronary arteries 1 week following the recombinant adenovirus vector infussion. The peak value of β galactosidase gene expression were detected in both the myocardium and coronary arteries 2 weeks after the vectors administration, which were 45.57 % in the myocardium and 26.11 % in the coronary arteries. The β gal expression was decreased 4 weeks later, and recovered to baseline at the end of eighth week.Conclusion:The experimental study indicates that efficiency of recombinant adenovirus mediated β gal gene expression was high in both the myocardium and coronary arteries. The approach of intracoronary infusion will be effective and feasible for gene therapy of coronary artery disease.更多还原