外源性基因在血管球囊损伤后新生内膜中的表达

In Vivo Gene Transfer and Expression in Thickening Neointima of Balloon-injured Blood Vessels

目的：应用重组腺病毒载体介导的β-半乳糖苷酶基因（LacZ）标记基因，探讨外源性基因在股动脉血管球囊损伤后，新生内膜中的表达。方法：用犬的股动脉内皮损伤模型，4周后经犬颈外动脉插管至股动脉内皮损伤处，并导入重组腺病毒载体介导的LacZ标记基因。转基因7天后，处死动物。β-半乳糖苷酶表达由x-Gal染色评价。经显微镜观察：在血管球囊损伤后，新生内膜中所见的蓝染细胞即为β-半乳糖苷酶表达阳性。结果：β-半乳糖苷酶表达出现在新生内膜、部分中膜和部分修复的内皮细胞。血管损伤后，新生内膜中平滑肌细胞转染率为100％。对照血管和远处脏器未显示β-半乳糖苷酶的表达。结论：在重组腺病毒载体的介导下，外源性基因可以在新生内膜中高效率地表达，新生内膜中的平滑肌细胞为基因治疗的靶细胞。

Objective: To study the expression of exogenous gene in thickening neointima of balloon-injured blood vessels,we transferred the marker gene LacZ to balloon-injured femoral artery of dogs through the mediation of recombinant adenovirus.Methods:At first,a balloon-injured femoral artery model of dogs was made. Four weeks later,we inserted the doubleballoon catheter through external carotid artery to injured sites of femoral artery. While the balloon was inflatedadenovirusmediated LacZ (Adex1SRLacZL) gene was transferred to the injured vessels. Seven days later, the anima is were sacrificed.x-Gal blue stain was used to evaluate the expression of fygalactosidase. The specimen was observed by microscope,the bluestain represents positive results.Results:The expression of Pgalactosidase appears in thickening neointima,some media and some repaired endothelium.cells. It is expressed in 100% of smooth muscle cells of the neointima,while the control group and distant organs has none ofthe expression of β-galactosidase.Conclusion: Through the mediation of recombinant adenovirus,Adex1SRLacZL gene can be expressed with high transferring efficiency in thickening neointima,which is good target cells for gene therapy.