微小RNA-29a及细胞基质金属蛋白酶2在脓毒症中的作用

Effects of microRNA-29a and matrix metalloproteinase-2 on sepsis

目的探讨微小RNA(miR)-29a及细胞基质金属蛋白酶2(MMP-2)在脓毒症中的作用。方法 (1)纳入脓毒症患者(脓毒症组)及健康者(对照组)各40例,检测其血清miR-29a和MMP-2表达水平,分析脓毒症患者血清miR-29a和MMP-2水平的相关性。应用TargetScan7. 1及miRanda软件预测MMP-2靶基因。(2)取对数生长期的RAW 264. 7细胞分为miR-29a对照物组和miR-29a模拟物组,转染后24 h、48 h、72 h、96 h检测细胞增殖情况。(3)取浓度为0μg/ml、1μg/ml、5μg/ml、10μg/ml脂多糖分别刺激RAW 264. 7细胞24 h,检测MMP-2蛋白的表达水平。(4)取RAW 264. 7细胞分为脂多糖组、脂多糖+miR-29a对照物组、脂多糖+miR-29a抑制物组和脂多糖+miR-29a模拟物组,转染6 h后加入10μg/ml脂多糖刺激24 h,检测MMP-2蛋白的表达。结果 (1)与对照组比较,脓毒症组血清miR-29a表达水平降低而MMP-2蛋白表达水平升高(P <0. 05)。脓毒症组miR-29a和MMP-2水平呈负相关(P <0. 05)。(2)转染后72 h miR-29a模拟物组的细胞增殖水平低于miR-29a对照物组(P <0. 05)。(3) 0μg/ml脂多糖、1μg/ml脂多糖、5μg/ml脂多糖、10μg/ml脂多糖刺激下RAW 264. 7细胞MMP-2蛋白相对表达水平依次升高(均P <0. 05)。(4)与脂多糖组和脂多糖+miR-29a对照物组相比,脂多糖+miR-29a模拟物组MMP-2表达降低,脂多糖+miR-29a抑制物组MMP-2表达升高(均P <0. 05)。结论 MMP-2及miR-29a均与脓毒症的发生密切相关,miR-29a可能通过调控MMP-2水平参与脓毒症的进程。

Objective To investigate the effects of microRNA(miR)-29a and matrix metalloproteinase-2(MMP-2) on sepsis.Methods (1)Forty patients with sepsis(sepsis group) and 40 healthy individuals(control group) were enrolled,and the serum miR-29a and MMP-2 expression levels were detected,then the correlation between serum miR-29a and MMP-2 levels was analyzed.The target genes of MMP-2 were predicted using TargetScan7.1 software and miRanda software.(2)The RAW 264.7 cells at logarithmic growth phase were collected and divided into miR-29a NC group oRmiR-29a mimic group,then the proliferation of the cells was detected 24 h,48 h,72 h and 96 h afteRtransfection separately.(3)The RAW 264.7 cells were stimulated by lipopolysaccharides(LPS) with the concentrations of 0 μg/ml,1 μg/ml,5 μg/ml and 10 μg/ml foR24 h separately,then the expression level of MMP-2 protein was detected.(4)The RAW 264.7 cells were divided into LPS group,LPS+miR-29a NC group,LPS+miR-29 ainhibitoRgroup,oRLPS+miR-29a mimic group.The cells were stimulated by 10 μg/ml LPS 6 h afteRtransfection foR24 h,then the expression level of MMP-2 protein was detected.Results (1)The serum miR-29a expression level decreased but MMP-2 protein expression level increased in the sepsis group than in the control group( P <0.05).The miR-29a level negatively correlated with MMP-2 level in the sepsis group( P <0.05).(2)72 h afteRtransfection,the cell proliferation of the miR-29a mimic group was loweRthan that of the miR-29a NC group( P <0.05).(3)The related expression level of MMP-2 protein increased successively in the cells stimulated by 0 μg/ml LPS,1 μg/ml LPS,5 μg/ml LPS and 10 μg/ml LPS(all P <0.05).(4)Compared to LPS group and LPS+miR-29a NC group,LPS+miR-29a mimic group had a loweRMMP-2 expression level ,while the LPS+miR-29a inhibitoRgroup had a higheRMMP-2 expression level(all P <0.05).Conclusion Both MMP-2 and miR-29a are closely related to the occurrence of sepsis,and miR-29a probably participates in the progress of sepsis by regulating MMP-2 level.