微小RNA-34a-5p通过靶向鼠双微体4抑制巨噬细胞氧化应激损伤的作用

Micro RNA-34a-5p inhibits oxidative stress damage in macrophages by targeting murine double minute 4

目的探讨微小RNA-34a-5p(miR-34a-5p)通过靶向鼠双微体4(MDM4)抑制巨噬细胞氧化应激损伤的作用。方法THP-1人单核细胞复苏后,培养分化为巨噬细胞。设立空白对照组。剩余细胞加入人源氧化低密度脂蛋白,孵育48 h,分化为巨噬泡沫细胞。设立动脉粥样硬化(AS)对照组。miR-34a-5p质粒转染,设为miR-34a-5p质粒转染组。每组24个重复。比较三组巨噬细胞凋亡情况,丙二醛(MDA)、超氧化物歧化酶(SOD)蛋白及活性氧(ROS)水平,MDM4阳性率。结果空白对照组凋亡率为(2.55±0.32)%,AS对照组凋亡率为(28.16±4.23)%,miR-34a-5p转染组为(13.48±1.66)%,三组比较差异有统计学意义(P<0.05)。AS对照组高于空白对照组与miR-34a-5p转染组,miR-34a-5p转染组高于空白对照组,差异有统计学意义(P<0.05)。三组MDA、SOD蛋白及ROS水平比较,差异均有统计学意义(P<0.05);AS对照组MDA、ROS水平最高,SOD水平最低,其次为miR-34a-5p转染组,组间比较差异均有统计学意义(P<0.05)。空白对照组MDM4阳性率为(8.33±0.35)%,AS对照组为(29.26±3.48)%,miR-34a-5p转染组为(14.26±1.59)%,三组比较差异有统计学意义(P<0.05)。结论miR-34a-5p可下调MDM4,进而抑制巨噬细胞氧化应激损伤。

Objective To investigate the inhibitory effect of micro RNA-34a-5p(miR-34a-5p)on oxidative stress damage of macrophages by targeting murine double minute 4(MDM4).Methods THP-1 human mononuclear cells were cultured and differentiated into macrophages after resuscitation.A blank control group was set up.The remaining cells were added to human oxidized low-density lipoprotein,incubated for 48 h,and differentiated into macrophage foam cells.AS control group was set up.miR-34a-5p plasmid transfection group was set as miR-34a-5p plasmid transfection.There were 24 replicates per group.The apoptosis of macrophages,the levels of MDA,SOD protein and ROS,and the positive rate of MDM4 were compared among the three groups.Results The apoptosis rate was(2.55±0.32)%in the blank control group,(28.16±4.23)%in the AS control group,and(13.48±1.66)%in the miR-34a-5p transfection group,the difference among the three groups was statistically significant(P<0.05).AS control group was higher than blank control group and miR-34a-5p transfection group,and miR-34a-5p transfection group was higher than blank control group,with statistical significances(P<0.05).There were statistically significant differences in MDA,SOD protein and ROS levels among the three groups(P<0.05).MDA and ROS levels were the highest and SOD levels were the lowest in AS control group,followed by miR-34a-5p transfection group,with statistical significances(P<0.05).MDM4 positivity rate was(8.33±0.35)%in the blank control group,(29.26±3.48)%in the AS control group,and(14.26±1.59)%in the miR-34a-5p transfection group,the difference among the three groups was statistically significant(P<0.05).Conclusion MiR-34a-5p can down-regulate MDM4 and inhibit the oxidative stress damage of macrophages.