LncRNA SNHG15靶向miR-370-3p调控ox-LDL诱导的血管内皮细胞损伤

LncRNA SNHG15 targets miR-370-3p to regulate ox-LDL-induced HUVECs cell injury

目的探讨长链非编码RNA(long noncoding RNA,lncRNA)小核仁RNA宿主基因15(Small nucleolar RNA host gene 15,SNHG15)对氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)细胞损伤的影响及其分子机制。方法100 mg/L ox-LDL处理HUVECs细胞24 h,记为ox-LDL组,未经ox-LDL处理的HUVECs细胞记为Con组。将si-NC、si-SNHG15、miR-NC、miR-370-3p分别转染至ox-LDL组,记为ox-LDL+si-NC、ox-LDL+si-SNHG15、ox-LDL+miR-NC和ox-LDL+miR-370-3p组;将si-SNHG15分别与anti-miR-NC、anti-miR-370-3p共转染至ox-LDL组,记为ox-LDL+si-SNHG15+anti-miR-NC和ox-LDL+si-SNHG15+anti-miR-370-3p组。实时荧光定量PCR(RT-qPCR)检测lncRNA SNHG15和miR-370-3p的表达量;流式细胞术和蛋白质印迹(Western blot)法分别检测细胞凋亡和蛋白表达;酶联免疫吸附试验检测白细胞介素-6(Interleukin-6,IL-6)、白细胞介素-1β(Interleukin-1β,IL-1β)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)水平。双荧光素酶报告实验检测lncRNA SNHG15和miR-370-3p的靶向关系。结果与Con组比较,经ox-LDL诱导的HUVECs细胞损伤中,lncRNA SNHG15和miR-370-3p的表达水平分别显著升高和降低(P<0.05)。抑制lncRNA SNHG15和过表达miR-370-3p均显著抑制细胞凋亡以及炎性细胞因子IL-6、IL-1β、TNF-α的表达(P<0.05)。lncRNA SNHG15靶向调控miR-370-3p表达,下调miR-370-3p逆转了抑制lncRNA SNHG15对ox-LDL引发凋亡以及炎性细胞因子的影响(P<0.05)。结论抑制lncRNA SNHG15通过靶向上调miR-370-3p缓解ox-LDL诱导的HUVECs细胞损伤。

Objective To investigate the effect of long non-coding RNA Small nucleolar RNA host gene 15(SNHG15)(lncRNA SNHG15)on oxidative low-density lipoprotein(ox-LDL)induced human umbilical vein endothelial cells(HUVECs)cell injury and its molecular mechanism.Methods HUVECs cells were treated with 100 mg/L ox-LDL for 24,named as ox-LDL group,and HUVECs cells untreated with ox-LDL were used as Con group.si-NC,si-SNHG15,miR-NC and miR-370-3p were transfected into ox-LDL group,named as ox-LDL+si-NC,ox-LDL+si-SNHG15,ox-LDL+miR-NC and ox-LDL+miR-370-3p group,respectively.si-SNHG15 with anti-miR-NC or anti-miR-370-3p were co-transfected into ox-LDL group,named as ox-LDL+si-SNHG15+anti-miR-NC and ox-LDL+si-SNHG15+anti-miR-370-3p group,respectively.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of lncRNA SNHG15 and miR-370-3p.Cell apoptosis and protein levels were assessed using Flow cytometry and Western blot,respectively.Enzyme-linked immunosorbent assay was used to evaluate the levels of Interleukin(IL)-6,IL-1βand Tumor necrosis factor-α(TNF-α).The targeted relationship between lncRNA SNHG15 and miR-370-3p was detected by double luciferase reporting experiment.Results Compared with the Con group,the expression levels of lncRNA SNHG15 and miR-370-3p were significantly increased and decreased in ox-LDL-induced HUVECs cell injury,respectively(P<0.05).Inhibition of lncRNA SNHG15 and overexpression of miR-370-3p significantly reduced apoptosis and the expression of inflammatory cytokines IL-6,IL-1βand TNF-α(P<0.05).LncRNA SNHG15 targeted the expression of miR-370-3p,and down-regulation of miR-370-3p reversed the effect of inhibition of lncRNA SNHG15 on ox-LDL-induced apoptosis and the expression of inflammatory cytokines IL-6,IL-1βand TNF-α(P<0.05).Conclusion Inhibition of lncRNA SNHG15 alleviates ox-LDL-induced HUVECs cell injury through targeted up-regulating miR-370-3p expression.