hCAP-18/LL-37基因转染对RAW264.7细胞活化功能的影响

Effect of hCAP-18/LL-37 Gene Transfection on Activation of RAW264.7 Cells

目的探讨抗菌肽hCAP-18/LL-37基因转染对巨噬细胞RAW264.7活化功能的影响。方法将重组质粒pcD-NA4/Myc-His-hCAP-18/LL-37瞬时转染RAW264.7细胞,MTT法检测细胞的增殖活性;中性红吞噬试验检测细胞的吞噬能力;RT-PCR法分析细胞活化相关分子CD80、CD86、TLR4及细胞因子IL-1β、TNF-αmRNA的转录。结果 hCAP-18/LL-37基因转染可促进经脂多糖(Lipopolysaccharide,LPS)处理的RAW264.7细胞的增殖活性与吞噬能力(P<0.05);可上调RAW264.7细胞CD80、CD86、TLR4、IL-1β、TNF-α基因mRNA的转录水平。结论 hCAP-18/LL-37基因转染可通过促进RAW264.7细胞增殖活性、吞噬能力及活化相关分子表达,调控巨噬细胞的活化功能。

Objective To investigate the effect of hCAP-18 / LL-37 gene transfection on the activation of macrophage RAW 264.7.Methods RAW264.7 cells were transiently transfected with recombinant plasmid pcDNA4 / Myc-His-hCAP-18 / LL-37,then determined for proliferation activity by MTT method,for phagocytosis ability by neutral red phagocytosis test,and for transcriptions of mRNAs of cell activation-related molecules CD80,CD86 and TLR4 as well as cytokines IL-1β and TNF-α.Results The transfection with hCAP-18 / LL-37 gene enhanced the proliferation activity and phagocytosis ability of RAW264.7 cells after treatment with lipopolysaccharide（LPS）（P 0.05）,and up-regulated the transcriptions of CD80,CD86,TLR4,IL-1β and TNF-α mRNAs.Conclusion The transfection with hCAP-18 / LL-37 gene regulated the activation of macrophage by enhancing the proliferation activity and phagocytosis ability of RAW264.7 cells as well as the expressions of activation-related molecules.