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目的:研究在细菌脂多糖(lipopolysaccharide,LPS)刺激肥大细胞炎症因子分泌过程中miRNA的表达差异。方法:肥大细胞P815培养后,用LPS(1μg/ml)激发肥大细胞,16 h后终止反应,通过酶联免疫吸附实验(ELISA)检测上清中肿瘤坏死因子α(TNF-α)、白介素-6(IL-6)的水平。再收集细胞,通过实时荧光定量PCR法检测miRNA的改变。结果:在LPS刺激肥大细胞后,TNF-α、IL-6的分泌与基础分泌量相比明显升高,差异有统计学意义。同时在肥大细胞中干预组miRNA-126的表达是正常组的1.3倍(P<0.05),干预组miRNA-155的表达是正常组的1.2倍(P<0.05),干预组miRNA-223的表达是正常组的1.7倍(P<0.05),干预组miRNA-221的表达是正常组的1.2倍(P<0.05),干预组miRNA-192的表达是正常组的1.4倍(P<0.05)。结论:miRNA在炎症因子LPS刺激肥大细胞P815后存在差异性表达,而这些差异表达的miRNA可能与肥大细胞的功能调节存在相关性。
Objective: To study the miRNA expression in the process of stimulating the secretion of inflammatory cytokines by bacterial lipopolysaccharide (LPS). Methods: Mast cells were stimulated with LPS (1μg / ml) and then stopped after 16 h. The expression of tumor necrosis factor - α (TNF - α), interleukin - 6 (IL-6) levels. Cells were harvested and miRNA changes detected by real-time fluorescence quantitative PCR. Results: After LPS stimulated mast cells, the secretion of TNF-α and IL-6 was significantly increased compared with the basal secretion, the difference was statistically significant. At the same time, the expression of miRNA-126 in the intervention group was 1.3-fold more than that in the normal group (P <0.05), and the expression of miRNA-155 in the intervention group was 1.2 times that of the normal group (P <0.05) (P <0.05). The expression of miRNA-221 in the intervention group was 1.2 times more than that in the normal group (P <0.05). The expression of miRNA-192 in the intervention group was 1.4 times more than that in the normal group (P <0.05). CONCLUSIONS: miRNAs are differentially expressed in the mast cells P815 stimulated by inflammatory cytokines LPS, and these differentially expressed miRNAs may be related to the functional regulation of mast cells.