腺病毒介导性基因导入血管平滑肌细胞

Adenovirus-mediated gene transfer into vascular smooth muscle cells

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 。

Objective:The efficiency of adenovirus mediated gene transfer into canine arterial smooth muscle cell (SMC) in culture were investigated.Method:A replication defective adenovirus expressing bacterial LacZ (Adexl SRLacZ)was introduced into SMC and β galatosidase activity in cell lysates was quantified.Result:The LacZ expression could be detectable by even a few minutes incubation with AdexlSRLacZ and gradually increased with the exposure time up to 2 hours.A dose dependent increase of LacZ expression was maintained up to moi 100,and the expression reached submaximal at moi 100.The time course of LacZ expression showed gradual increase and a plateau in 3 to 5 days.The gene transduction by AdexlSRLacZ induced more than 250 times higher LacZ expression than that by lipofection.Almost 100% of cells exposed to AdexlSRLacZ showed characteristic blue staining with X gal,which only a few percentage of cells subjected to DNA lipofection complexes were stained under microscopic examination.Conclusion:The results indicated that adenovirus mediated gene transfer showed rapid infection kinetics and induced high efficiency of LacZ expression in SMC.