微小RNA-4804对人单核细胞株THP-1细胞增殖与分泌细胞因子的影响研究

Effects of mi RNA-4804 on proliferation and cytokines secreted function in human monocyte line

目的探讨miRNA-4804对人单核细胞增殖和分泌细胞因子功能的影响。方法实时定量PCR检测人单核细胞株THP-1中miRNA-4804和细胞因子m RNA的表达,流式细胞仪与CCK8检测miRNA-4804对THP-1细胞增殖的影响,酶联免疫吸附测定THP-1细胞培养上清中细胞因子的表达。结果与对照组比较,加入miRNA-4804模拟剂,IFN-γ诱导的THP-1细胞增殖、HLA-DR、CD86分子的表达明显受到抑制,CD80表达无明显变化;而加入miRNA-4804抑制剂,THP-1细胞的增殖,HLA-DR、CD86分子的表达明显增加,CD80表达也明显增加。IFN-γ刺激后,THP-1细胞和培养上清中IL-6、TNF-α和IL-12的表达明显增强,IL-10的表达明显下降;miRNA-4804模拟剂处理后,能拮抗IFN-γ诱导THP-1细胞表达IL-6、TNF-α和IL-12的作用,且促进THP-1细胞表达IL-10。反之,miRNA-4804抑制剂处理后,能促进IFN-γ诱导THP-1细胞表达IL-6、TNF-α和IL-12的作用,而抑制THP-1细胞表达IL-10,差异均具有统计学意义(P<0.05)。结论 miRNA-4804能够影响单核细胞的增殖和分泌细胞因子的功能,可能在慢性乙型肝炎免疫稳态机制中发挥作用。

Objective To investigate the effect of miRNA-4804 on proliferation and cytokines secreted function of human monocyte line. Methods The expression of miRNA-4804 and cytokine m RNA in human monocytic cell line THP-1 were detected by real-time quantitative PCR. Detection of THP-1 cell proliferation and surface molecule were used by flow cytometry and CCK8. The expression of cytokines in the supernatant of cell culture were detected by enzyme linked immunosorbent assay. Results Compared with control group, the proliferation and expression of HLA-DR, CD86 molecules of THP-1 cell induced with IFN-γ were significantly inhibited by the miRNA-4804 agonist. The expression of CD80 had no obvious change. THP-1 cell proliferation and the expression of HLA-DR, CD86, CD80 molecules were significantly increased by the addition of the miRNA-4804 inhibitor. The expression of IL-6, TNF-α and IL-12 increased, IL-10 significantly decreased in culture supernatant of THP-1 cells upon IFN-γ stimulation. The expression of IL-6, TNF-α and IL-12 in THP-1 cells induced with IFN-γ were antagonistic after treatment with miRNA-4804 mimics. The expression of IL-10 in THP-1 cells were promoted. On the contrary, the expression of IL-6, TNF-α and IL-12 in THP-1 cells induced with IFN-γ were promoted and IL-10 were inhibited after treatment with miRNA-4804 inhibitor. Conclusion miRNA-4804 can influence the proliferation and cytokine secretion of monocytes and might be involved in the regulation of the immune homeostasis of chronic hepatitis B.