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目的 :探讨腺病毒介导性基因导入血管平滑肌细胞 ( SMC)的有效性。方法 :选用含有细菌 L ac Z基因的重组腺病毒及脂质体载体导入培养主动脉 SMC并用组织化学法定性定量检测基因表达的产物。结果 :腺病毒与 SMC接触几分钟就可使 SMC转染 ,2 h即可使 10 0 %的细胞受其转染并表达 L ac Z基因 ;基因表达的程度与病毒滴度呈剂量依赖关系 ;转染后 2 4h即可在 SMC中检出外源基因的表达 ,3~ 5 d达高峰 ;脂质体转导法仅可使不足 1%的 SMC表达外源基因 ;腺病毒介导性基因导入法约为脂质体转导法的 2 5 0倍。结论 :本研究获得的腺病毒介导性基因导入 SMC的多种参数 ,可为其未来的研究和治疗应用提供依据
Objective: To investigate the effectiveness of adenovirus-mediated gene transfer into vascular smooth muscle cells (SMCs). Methods: The recombinant adenovirus containing Lactobacillus plantarum and liposome were introduced into cultured aortic SMC and qualitatively and quantitatively detected the gene expression products by histochemistry. Results: Adenovirus could be transfected into SMC in a few minutes after contact with SMC. After 2 h, 100% of cells were transfected with Lac Z gene. The extent of gene expression was dose-dependent with virus titer. The expression of exogenous gene was detected in SMC 24 h after transfection, reaching the peak at 3 ~ 5 d. The liposome transduction method only allowed the expression of foreign genes in less than 1% of SMCs. The adenovirus-mediated gene The introduction method is about 25O times of the liposome transduction method. Conclusion: The various parameters of adenovirus-mediated gene transfer into SMC obtained in this study may provide the basis for its future research and therapeutic application