紫铆花素通过下调miR-34a对氧化低密度脂蛋白诱导血管内皮细胞损伤的影响

Effects of butein on oxidized low-density lipoprotein-induced vascular endothelial cell injury by down-regulating miR-34a

目的探讨紫铆花素对氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉血管内皮细胞(HUVECs)损伤的影响。方法体外培养的HUVECs,将其分为对照组、模型组、低、中、高剂量紫铆花素组、anti-miR-NC组、anti-miR-34a组、高剂量紫铆花素加miR-NC组、高剂量紫铆花素加miR-34a组;分别进行RT-qPCR、流式细胞术和Western blot检测miR-34a表达水平、细胞凋亡率和相关蛋白表达;ELISA检测白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平。结果与对照组比较,模型组细胞凋亡率、炎性细胞因子IL-6、IL-1β和TNF-α表达水平均显著升高(P<0.05);紫铆花素处理或下调miR-34a均可显著降低细胞凋亡率以及炎性细胞因子IL-6、IL-1β和TNF-α的表达水平(P<0.05);紫铆花素可显著降低miR-34a的表达水平(P<0.05);且上调miR-34a逆转了紫铆花素对ox-LDL处理HUVECs凋亡和炎性细胞因子的影响。结论紫铆花素可能通过下调miR-34a表达缓解ox-LDL诱导的HUVECs损伤。

Objective To investigate the effect of butein on the injury of human umbilical vein endothelial cells(HUVECs)induced by oxidized low-density lipoprotein(ox-LDL).Methods HUVECs were cultured in vitro and divided into control group,model group,low,medium and high dose butein group,anti-miR-NC,anti-miR-34a,high dose butein combined with miR-NC group,high dose butein combined with miR-34a group.RT-qPCR,flow cytometry and Western blot were used to detect the expression of miR-34a,apoptosis and the expression of related proteins.ELISA was used to evaluate the levels of interleukin-6(IL-6)、interleukin-1β(IL-1β)、tumor necrosis factor-α(TNF-α).Results Compared with the control group,the apoptosis rate and the expression levels of inflammatory cytokines IL-6,IL-1βand TNF-αin model group significantly increased(P<0.05).Butein treatment or down-regulation of miR-34a expression could significantly reduce the apoptosis rate and the expression levels of inflammatory cytokines IL-6,IL-1βand TNF-α(P<0.05).Butein could significantly reduce the expression level of miR-34a(P<0.05).Up-regulation of miR-34a reversed the effects of butein on the apoptosis and inflammatory cytokines of ox-LDL-treated HUVECs.Conclusion Butein may alleviate ox-LDL-induced HUVECs injury by down-regulating miR-34a expression.