【摘要】 目的探讨共聚物P85与黏附增强型绿色荧光蛋白(enhanced green fluoresent protein,EGFP)质粒的微泡造影剂(microbubble,MB)结合后联合一定强度的超声(ultrasound,US)辐照,是否增强人肝癌细胞(HepG2)的质粒转染及表达。方法以人肝癌细胞(HepG2)为研究对象,质粒DNA是可表达绿色荧光蛋白的pEGFP,添加微泡造影剂或共聚物P85后进行脉冲多普勒超声辐照(频率1MHz,声强1W/cm~2,工作周期20%,时间20s)。24h后台盼蓝染色评价细胞存活率,荧光显微镜和流式细胞仪评价细胞的基因转染率。结果有超声辐照组的pEGFP转染率明显高于无超声辐照组(P<0.01),有超声辐照组中pEGFP+30%MB+P85+US组转染效率(22.14±3.06)%优于单独使用微泡组(11.34±2.2)%或P85组(9.72±1.21)%(P<0.01)。结论微泡造影剂与共聚物P85结合同时给予超声辐照可增强基因转染效率,在基因治疗上有待于进一步关注和研究。更多还原

【Abstract】 Objective To investigate whether Pluronic P85 and plasmid EGFP which adhered on the microbubble (MB) could enhance the gene transfection and expression to HepG2 cell under ultrasound(US) irradiation.Methods Plasmid encoding enhanced green fluorescent protein(pEGFP) was used as a report gene.HepG2 cell and plasmids DNA with or without microbubble/P85 were exposed to ultrasound(US parameters:1 MHz,1.0 W/cm~2,20 s,20% duty cycle) and after 24 h transfection rates and cell viability were assessed by fluorescence microscopy,fluorescence activated cell sorting(FACS) analysis,and trypan blue exclusion.Results The gene transfection to HepG2 cell under ultrasound irradiation was much higher compared to that without ultrasound irradiation.When they were exposed to ultrasound,microbubble alone or P85 alone expressed lower green fluorescent protein,microbubble plus P85 obtained the higher expression efficiency at 30%microbubble concentration without obvious decreasing of survival rate(55.73±3.32)%.Conclusions Microbubble with pluronic P85 could enhance the gene transfection under ultrasound irradiation, and it could deserve further attention and investigation.更多还原