目的 探讨miRNA-24-3P(miR-24-3P)在急性呼吸窘迫综合征(ARDS)过程中的作用机制.方法 在RAW264.7细胞中分别转染miR-24-3P mimics、miR-24-3P inhibitor及各自阴性对照,检测各组细胞中miR-24-3P的相对表达水平,检测各组细胞中肿瘤坏死因子-a(TNF-α)、白细胞介素-6(IL-6)、SP1、核因子-κB(NF-κB)的相对表达水平,并通过双荧光素酶报告基因检测系统验证miR-24-3P对SP1的靶向作用.结果 TNF-α mRNA、IL-6 mRNA在各组细胞中的表达水平差异有统计学意义(P＜0.05).SP1 mRNA、NF-κBmRNA在各组细胞中的相对表达水平差异无统计学意义(P＞0.05).SP1在各组细胞中的表达水平差异有统计学意义(P＜0.05).NF-κB在各组细胞中的相对表达水平差异无统计学意义(P＞0.05).双荧光素酶报告基因分析表明,共同转入miR-24-3P与SP1野生型会显著抑制HEK293细胞中的荧光酶表达(P＜0.05).结论 miR-24-3P可通过影响SPI基因的翻译进而影响炎性介质的释放,在ARDS过程中发挥作用.%Objective To discuss the mechanism of miR-24-3P in the process of acute respiratory distress syndrome (ARDS).Methods miR-24-3P mimics,miR-24-3P inhibitor and their negative controls were transfected in RAW264.7 cells respectively:The relative expression of miR-24-3P in each cell was detected.The relative expression of TNF-α,IL-6,SP1 and NF-κB were detected in each cell,and the regulation of miR-24-3P on the target gene SP1 was detected by dual luciferase reporter gene.Results The expression level of TNF-α mRNA and IL-6 mRNA in each group was statistically significant (P＜0.05).The relative expression level of SP1 mRNA and NF-κB mRNA in each group was not statistically significant (P＞0.05).The expression of SP1 in each group was statistically significant (P＜0.05).The gene analysis of dual luciferase reporter showed that the fluorescence activity was significantly inhibited after cotransfection of miR-24-3P and SP1 in HEK293 cells(P＜0.05).Conclusion miR-24-3P can play an important role in the process of ARDS by influencing the translation of SP1 gene and affecting the release of inflammatory mediators.
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