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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 the function of dendritic cells and subsets induced by lipopolysaccharide. Methods miRNA let-7i inhibitor was used to intervene in LPS-induced DC maturation. The expression of miRNA let-7i was detected by RT-PCR. The phenotype of DC was analyzed by flow cytometry. The let- 7i inhibitor-treated DCs were divided into two groups: CD86 + DC and CD86-DC, which were respectively co-cultured with T cells to observe the effect of CD86 + DC and CD86-DC on T cell proliferation; CD86 + DC and CD86- DC Secretion of cytokines IL-10, IL-12 and TNF-α levels. Results Transfection of miRNA let-7i inhibitor significantly reduced the expression of CD80 and CD86 on the surface of LPS-stimulated DCs. LPS-stimulated DCs can be divided into two subgroups with down-regulation of let-7i: CD86 + DC and CD86-DC, and CD86-DC can induce more regulatory T cells and anti-inflammatory cytokines more effectively Pro-inflammatory cytokines decrease. Conclusion Inhibition of miRNA let-7i induces the expression of CD86-DCs in DCs, leading to immune tolerance.