胆固醇衍生物阳离子脂质体复合EGFP—IL-1Ra质粒体外转染HELA细胞系的研究

目的研究胆固醇衍生物阳离子脂质体复合EGFP—IL—IRa质粒体外转染HELA细胞系的效率，为类风湿性关节炎治疗提供新的选择。方法制备新型胆固醇衍生物脂质体及EGFP—IL-1Ra质粒，两者结合转染HELA细胞系，通过体外凝胶阻滞试验检测阳离子脂质体与质粒DNA的最佳配比关系，MTT法检测脂质体细胞毒性，荧光免疫法检测HELA细胞的转染效率。结果实验中所用脂质体的IC50为650μg／mL，较对照组有更低的细胞毒性。脂质体／DNA质量比为50：1作为最适比例，荧光显微镜观测脂质体结合EGFP—IL-1Ra质粒转染HELA荧光强度高于单纯EGFP—IL-1Ra质粒转染组。结论胆固醇衍生物阳离子脂质体复合EGFP—IL-1Ra质粒可高效的转染HELA细胞系，有望成为类风湿性关节炎治疗的新方法。

The transgene effect of cholesterol derivative-cationic liposome EGFP-IL-1Ra plasmid complex in HELA cell line

Objective To observe the transgene effect of cholesterol derived cation liposome combine EGFP-IL-1Ra plasmid in HELA cell line,exploring new therapeutic methods for the treatment of rheumatoid arthritis.Methods Cholesterol derived cation liposome as well as EGFP-IL-1Ra plasmid were prepared and combined to transfer the HELA cell line.Gel retarding was performed to select the appropriate liposome/DNA ratio,MTT was used to observe the cell toxicity of this complex.The transfer efficiency was observed using fluorescence microscope.Results The IC50 of theliposome was 650 μg/mL with less cell toxicity compared with the control.The 50 ∶1 was the best liposome/NDA ratio for transferation and the cholesterol derived cation liposome complex group inidicated higher fluorescence intensity compared with the EGFP-IL 1Ra loposome group.Conclusion Cholesterol derived cation liposome combine EGFP-IL-1Ra plasmid can tranfer the HELA cell line with high efficiency,which has potential therapeutic effect on RA.