MicroRNA let-7i对脂多糖诱导的树突状细胞及亚群功能的影响  

Effect of inhibiting miRNA let-7i on the subsets and function of LPS-induced DC

作　　者：李立群[1] 吴健[2,3] 张毛毛[2,3] 刘芳[2,3] 韩晓丽[2,3] 谭妙欣[2,3] 史呈伟[1] 刘心平[1] 王菲[1] 于丹丹[1] 于波[2,3] 

机构地区：[1]哈尔滨医科大学科研处 [2]哈尔滨医科大学附属第二医院心血管内科 [3]哈尔滨医科大学心肌缺血机理与诊疗技术教育部共建重点实验室,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2012年第1期5-9,共5页Journal of Harbin Medical University

基　　金：黑龙江省教育厅资助课题(11551234);黑龙江省青年科学基金资助(QC2010048);哈尔滨市科技局科技创新人才研究专项资金(2009RFQXS226)

摘　　要：目的探讨miRNA let-7i对脂多糖诱导的树突状细胞及亚群功能的影响。方法在LPS诱导DC成熟过程中,用miRNA let-7i抑制剂进行干预,然后用RT-PCR检测miRNA let-7i的表达水平;流式细胞仪分析DC的表型;用免疫磁珠将let-7i抑制剂处理的DCs分成两组:CD86+DC和CD86-DC,分别与T细胞共培养,观察CD86+DC和CD86-DC对T细胞增殖的影响;通过ELISA检测CD86+DC和CD86-DC分泌的细胞因子IL-10,IL-12和TNF-α的水平。结果转染miRNA let-7i抑制剂明显降低了LPS刺激的DC表面CD80和CD86的表达。LPS刺激的DCs伴随let-7i下调可分为两个亚群:CD86+DC和CD86-DC,且CD86-DC能更有效地诱导调节性T细胞的增多和抗炎症细胞因子的增多,并使促炎症细胞因子减少。结论抑制miRNA let-7i可以诱导DCs中CD86-DCs的表达,进而导致免疫耐受。Objective To investigate the effect of miRNA let-7i on CD86 - DC subpopulations of DC stimulated by lipopolysaceharide（LPS）. Methods miRNA let-7i inhibitor was used to in- terfere LPS-indueed DC maturation, the levels of miRNA let-7i in cocuhure system were detec- ted by RT-PCR; DC phenotype were analyzed by using flow cytometry during LPS-induced DC maturation;miRNA let-Ti inhibitor transfeeted DC were divided into CD86 ＋ DCs and CD86- DCs by using immunomagnetic beads, cocultured with T cells and observed the effect of CD86 ＋ DC and CD86 - DC on T cell proliferation. The levels of IL-10, IL-12, and TNF-α secreted byCD86 ＋ DC and CD86- DC were detected by ELISA. Results Transfecting with miRNA let-7i inhibitor significantly impeded expression of CDS0 and CD86 on DC surface and the secretion of pro-inflammatory cytokine. There were two subpopulations of LPS-stimulated DCs with down- regulated miRNA let-7i, CD86 ＋ DC and CD86 - DC, and it was the CD86 - DCs that were more effective in inducing Treg cell increase and the secretion of anti-inflammatory increase, and re- duced the secretion of pro-inflammatory cytokine. Conclusion Inhibiting miRNA let-7i can induce the expression of CD86 - DCs in the overall DCs group, thereby leading to immune toler- ance.

关 键 词：树突状细胞 miRNA let-7i T细胞 细胞因子 

分 类 号：R392.1[医药卫生—免疫学]