miR-145-5p通过靶向DUSP6抑制LPS诱导的血管内皮细胞氧化损伤和炎症

miR-145-5p inhibits LPS-induced oxidative damage and inflammation in vascular endothelial cells by targeting DUSP6

目的:研究miR-145-5p对脂多糖(LPS)诱导的血管内皮细胞氧化损伤和炎症的调节及相关机制。方法:LPS诱导的人脐静脉内皮细胞(HUVECs)作为炎症损伤模型。分为Control组,LPS组,LPS+mimic组,LPS+DUSP6组和LPS+mimic+DUSP6组。采用RT-qPCR检测细胞中miR-145-5p和DUSP6表达;流式细胞术检测细胞凋亡,Western blot检测Bax/Bcl-2比率;试剂盒检测氧化应激;RT-PCR结合ELISA检测细胞上清液中的炎症因子IL-10和iNOS水平。结果:miR-145-5p与DUSP6具有良好的靶向关系。与Control组相比,LPS组细胞中DUSP6表达显著升高(P<0.05)。与LPS组相比,LPS+DUSP6组中Bax/Bcl-2比率均显著升高,SOD含量显著降低而MDA含量与iNOS相对mRNA水平显著升高(P<0.05)。而与LPS+DUSP6组相比,LPS+mimic+DUSP6组中SOD与IL-10含量显著升高,而Bax/Bcl-2比率,MDA含量及iNOS相对mRNA水平均显著下降(P<0.05)。结论:miR-145-5p过表达可通过靶向DUSP6缓解LPS诱导的血管内皮细胞氧化损伤和炎症。

Objective:To study the regulation of miR-145-5p on oxidative damage and inflammation of vascular endothelial cells induced by lipopolysaccharide(LPS)and its related mechanisms.Methods:Human umbilical vein endothelial cells(HUVECs)induced by LPS were used as a model of inflammatory injury.Control group,LPS group,LPS+mimic group,LPS+DUSP6 group and LPS+mimic+DUSP6 group.RT-qPCR was used to detect the expression of miR-145-5p and DUSP6 in cells,flow cytometry was used to detect apoptosis,Western blot was used to detect the ratio of Bax/Bcl-2;kit was used to detect oxidative stress;RT-PCR combined with ELISA were used to detect cell supernatant levels of inflammatory factors IL-10 and iNOS in serum.Results:miR-145-5p has a good targeting relationship with DUSP6.Compared with the control group,the expression of DUSP6 in the cells of the LPS group was significantly increased(P<0.05).Compared with the LPS group,the ratio of Bax/Bcl-2 in the LPS+DUSP6 group was significantly increased,the SOD content was significantly reduced,MDA content and the relative mRNA levels of iNOS were significantly increased(P<0.05).However,compared with the LPS+DUSP6 group,contents of SOD and IL-10 in the LPS+mimic+DUSP6 group were significantly increased,the ratio of Bax/Bcl-2,MDA content and iNOS relative mRNA levels were significantly reduced(P<0.05).Conclusion:miR-145-5p overexpression can alleviate oxidative damage and inflammation of vascular endothelial cells induced by LPS by targeting DUSP6.