沉默 HOXA11-AS 靶向 miR-766-3p 对 ox-LDL 诱导的血管内皮细胞损伤的影响

目的 探讨长链非编码 RNA (lncRNA) 同源异形盒基因 A11 反义 RNA (HOXA11-AS) 靶向微小RNA-766-3p (miR-766-3p) 对氧化低密度脂蛋白 ( ox-LDL) 诱导的血管内皮细胞损伤的影响。 方法 以100 μg / mL 的 ox-LDL 处理人脐静脉血管内皮细胞 (human umbilical vein endothelial cell, HUVEC) 24 h 建立细胞损伤模型。 将 HUVEC 分为对照 (con) 组、 ox-LDL 组、 ox-LDL + si-NC 组、 ox-LDL + si-HOXA11-AS 组、ox-LDL + miR-NC 组、 ox-LDL + miR-766-3p 组、 ox-LDL + si-HOXA11-AS + anti-miR-NC 组、 ox-LDL + si-HOXA11-AS + anti-miR-766-3p 组。 RT-qPCR 检测 HOXA11-AS 和 miR-766-3p 表达。 流式细胞术分析细胞凋亡。试剂盒检测 LDH 释放量、 胞内 SOD 活性。 ELISA 法检测培养液中 TNF-α、 IL-1β 水平。 双荧光素酶报告实验确定 HOXA11-AS 和 miR-766-3p 的靶向关系。 结果 与 con 组比较, ox-LDL 组 HUVEC 细胞凋亡率、 LDH释放量、 HOXA11-AS 表达量以及培养液中 TNF-α、 IL-1β 水平显著升高 (P< 0. 05), SOD 活性、 miR-766-3p表达量显著降低 (P< 0. 05)。 与 ox-LDL + si-NC 组比较, ox-LDL + si-HOXA11-AS 组 HUVEC 细胞凋亡率、LDH 释放量以及培养液中 TNF-α、 IL-1β 水平显著降低 (P< 0. 05), SOD 活性显著升高 (P< 0. 05)。 与 ox-LDL + miR-NC 组比较, ox-LDL + miR-766-3p 组 HUVEC 细胞凋亡率、 LDH 释放量以及培养液中 TNF-α、 IL-1β 水平显著降低 (P< 0. 05), SOD 活性显著升高 (P< 0. 05)。 miR-766-3p 是 HOXA11-AS 的直接靶点。 与ox-LDL + si-HOXA11-AS + anti-miR-NC 组比较, ox-LDL + si-HOXA11-AS + anti-miR-766-3p 组 HUVEC 细胞凋亡率、 LDH 释放量以及培养液中 TNF-α、 IL-1β 水平显著升高 (P< 0. 05), SOD 活性显著降低 (P< 0. 05)。 结论 沉默 HOXA11-AS 通过靶向上调 miR-766-3p 表达能够抑制 ox-LDL 诱导的血管内皮细胞凋亡、 氧化应激和炎性反应。

ct of Silencing HOXA11-AS on ox-LDL-induced Vascular Endothelial Cell Injury by Targeting miR-766-3p

Objective To investigate the effect of long non-coding RNA ( lncRNA) homologous anomalous box gene A11 antisense RNA (HOXA11-AS) on oxidative low density lipoprotein(ox-LDL) induced vascular endothelial cells injury by targeting micrornA-766-3p (miR-766-3p). Methods Human umbilical vein endothelial cells (HUVEC) were treated with 100 μg / mL ox-LDL for 24 hours to establish a cell injury model. HUVEC were divided into the control ( con)group, ox-LDL group, ox-LDL + si-NC group, ox-LDL + si-HOXA11-AS group, ox-LDL + miR-NCgroup, ox-LDL + miR-766-3p group, ox-LDL + si-HOXA11-AS + anti-miR-NC group, and the ox-LDL + si-HOXA11-AS + anti-miR-766-3p group. Expression of HOXA11-AS and miR-7666-3p was assessed by RT-qPCR. Cell apoptosis was detected using flow cytometry. The kits detected LDH releaseand intracellular SOD activity. ELISA method was used to calculate the levels of TNF-α and IL-1β inthe culture medium. Dual luciferase reporter assay was used to confirm the targeting relationship between HOXA11-AS and miR-766-3p. Results Compared with the control group, the HUVEC cellapoptosis rate, LDH release, HOXA11-AS expression, and the levels of TNF-α and IL-1β in theculture medium of the ox-LDL group were significantly increased (P< 0. 05), the SOD activity andthe miR-766-3p expression level were significantly reduced (P< 0. 05). Compared with the ox-LDL+ si-NC group, the HUVEC cell apoptosis rate, LDH release, and the levels of TNF-α and IL-1βin the culture medium of the ox-LDL + si-HOXA11-AS group were significantly reduced (P< 0. 05),the SOD activity was significantly increased ( P < 0. 05). Compared with the ox-LDL + miR-NCgroup, the HUVEC cell apoptosis rate, LDH release, and the levels of TNF-α and IL-1β in theculture medium of the ox-LDL + miR-766-3p group were significantly reduced ( P < 0. 05), theSOD activity was significantly increased ( P < 0. 05 ). miR-766-3p is the a target of HOXA11-AS. Compared with the ox-LDL + si-HOXA11-AS + anti-miR-NC group, the HUVEC cell apoptosisrate, LDH release, and levels of TNF-α and IL-1β in the culture medium of the ox-LDL + si-HOXA11-AS + anti-miR-766-3p group were significantly increased (P< 0. 05), and the SOD activity was significantly decreased (P< 0. 05). Conclusion Silencing HOXA11-AS inhibits ox-LDL-induced vascular endothelial cell apoptosis, oxidative stress and inflammation by up-regulating the expression ofmiR-766-3p.