摘要
观察以多聚乙烯亚胺(polyethylenimine,PEI)为载体,介导活化并发生凋亡的淋巴细胞来源的DNA(凋亡DNA)的胞内递送,探讨凋亡DNA对原代巨噬细胞活化的影响。采用PEI为载体,介导凋亡DNA和正常淋巴细胞来源的DNA(正常DNA)进入胞内,以ELISA方法检测巨噬细胞IL-6、TNF-α的分泌;分析各种因素对PEI胞内递送效率的影响,并评价其对原代巨噬细胞的安全使用剂量。结果显示:PEI可介导DNA对原代巨噬细胞的胞内递送;凋亡DNA进入胞内后,可以刺激巨噬细胞活化,诱导炎症因子IL-6和TNF-α的分泌;相同的条件下,正常DNA却不能引起这些变化。进一步观察发现PEI与DNA的质量比为2或3时胞内递送效率最佳;小于10μg/ml的PEI用量并于转染6 h后换液对原代巨噬细胞毒性较小。上述结果表明,PEI作为载体,可以有效介导对原代巨噬细胞的基因(哺乳动物DNA或质粒)递送;进入胞内的凋亡DNA可促进巨噬细胞活化并分泌大量炎性细胞因子,这对于研究系统性红斑狼疮(SLE)的发病机制具有意义。
To explore the role of intracytoplasmic apoptotic DNA on the activation of primary macrophages, polyethylenimine （PEI）, a polycationic DNA carrier, was used to deliver apoptotic DNA into the cytoplasm of primary macrophages. The levels of IL-6 and TNF-α were quantified by ELISA assay, and the influence of various factors on the deliver efficiency of PEI was investigated to obtain the optimal working scheme. And the cytotoxicity of PEI was also evaluated. The experimental result showed that the highest delivery efficiency was achieved when PEI/DNA （w/w） ratio was 2 or 3 and no cytotoxicity was found with less than 10μg/ml PEI dosage which should be removed 6 hs after transfection. Intracytoplasmic delivery of apoptotic DNA induced production of abundances of IL-6 and TNF-α by the macrophage, while normal DNA failed to have the same effect. These results suggest that PEI is a useful vector for gene delivery in primary cells, and intraeytoplasmic apoptotic DNA has ability to activate the primary macrophages to produce inflammatory cytokines, which may facilitate our further understanding of the pathogenesis of systemic lupus erythematosus （SLE）.
出处
《现代免疫学》
CAS
CSCD
北大核心
2009年第2期95-99,共5页
Current Immunology
基金
国家自然科学基金资助项目(30671952)
上海市科委重点资助项目(07JC14004)
