体外实验观察微泡-阳离子纳米脂质体复合物促进HGF基因转染

目的通过生物素-亲和素的特异结合,制备一种高效载肝细胞生长因子(HGF)基因的脂质微泡(LMB)-阳离子纳米脂质体复合物(LMB-CNLP),观察转染情况,探讨其对对数生长期的肝星状细胞株HSC-T6活性的影响。方法将HSC-T6细胞分为载HGF基因的LMB组、LMB-CNLP组和对照组(单纯质粒),超声辐照各组进行HGF转染。以荧光显微镜及流式细胞仪观测转染情况,ELISA试剂盒检测HGF蛋白表达情况,流式细胞仪检测HSC-T6细胞凋亡。结果 LMB-CNLP组HGF基因的转染率为41.29%,明显高于其他2组。转染后的细胞能持续表达HGF,且随着HGF蛋白增多,HSC-T6细胞凋亡率增高。结论超声辐照LMB-CNLP复合物能提高HGF基因在体外培养的HSC-T6细胞中的转染情况,促进HSC-T6凋亡,为基因治疗肝纤维化提供实验依据。

In vitro combination of lipid microbubbles and cationic nanoliposomes in enhancing HGF gene transfection

Objective To combine lipid microbubbles with cationic nanoliposomes by biotin-avidin system as a type of gene HGF vector, in order to investigate the transfection and the cytoactive of hepatic stellate cell (HSC-T6). Methods HSC-T6 was randomly divided into LMB group (given lipid microbubble and plasmid), LMB-CNLP group (given loaded-gene combination of lipid microbubbles with cationic nanoliposomes) and contrast group (given a dose of plasmid). Expression of the gene HGF was observed on fluorescent microscopy and quantified with FACS analysis. ELISA was used to detect the expression of HGF. The viability of HSC-T6 was measured with FACS analysis. Results Transfection efficiency in LMB-CNLP group was 41.29%, higher than that in the other two groups. The HSC-T6 in LMB-CNLP group expressed HGF protein persistently. With HGF protein increased, more HSC-T6 apoptosis was observed. Conclusion Combination of lipid microbubbles with cationic nanoliposomes can enhance the efficiency of gene HGF transfection in HSC-T6 and promote cell apoptosis of HSC-T6, offering a experimental basis for the treatment of liver fibrosis.