乙醛脱氢酶2对单核细胞-人主动脉内皮细胞黏附的影响及相关机制研究

Effect of aldehyde dehydrogenase 2 on monocyte-human aortic endothelial cells adhesion and its mechanisms

摘要:

目的： 探讨乙醛脱氢酶2（aldehyde dehydrogenase 2，ALDH2）对单核细胞（human acute monocytic leukemia cells，THP-1）-人主动脉内皮细胞（human aortic endothelial cells，HAECs）黏附的影响及相关机制。方法： 将HAECs分为对照（Control）组，ALDH2 NC组，ALDH2组，siRNA-ALDH2 NC组，siRNA-ALDH2-1、-2、-3组，Western blot及real-time PCR检测转染效果，细胞黏附实验检测THP-1-HAECs黏附能力，Western blot及real-time PCR检测核转录因子-κB（nuclear factor-kappa B，NF-κB）转录情况。结果： 与Control组、ALDH2 NC组比较，ALDH2组中ALDH2 mRNA及蛋白表达量上调（P<0.05）；与Control组、siRNA-ALDH2 NC组比较，siRNA-ALDH2-1、-2、-3组中ALDH2 mRNA及蛋白表达量下调（P<0.05），其中siRNA-ALDH2-2组中ALDH2 mRNA及蛋白表达量最低。与Control组、ALDH2 NC组比较，ALDH2组中THP-1-HAECs黏附能力降低（P<0.05），细胞核中NF-κB表达量下调（P<0.05），细胞质中NF-κB表达量上调（P<0.05），红色荧光减弱（P<0.05）；与Control组、siRNA-ALDH2 NC组比较，siRNA-ALDH2组中THP-1-HAECs黏附能力提高（P<0.05），细胞核中NF-κB表达量上调（P<0.05），细胞质中NF-κB表达量下调（P<0.05），红色荧光增强（P<0.05）。结论： ALDH2通过抑制NF-κB核转录进而抑制THP-1-HAECs黏附，最终缓解动脉粥样硬化。

Abstract:

Objective: To investigate the effect of aldehyde dehydrogenase 2 (ALDH2) on monocyte (THP-1) -human aortic endothelial cells (HAECs) adhesion and its mechanisms. Methods: HAECs was divided into the Control group, ALDH2 NC group, ALDH2 group, siRNA-ALDH2 NC group, siRNA-ALDH2-1, -2, -3 groups. The expression of ALDH2 protein and mRNA was detected by Western blot and real-time PCR. The adhesive ability between THP-1 and HAECs was detected by cell adhesion assay. The transcription of nuclear factor-kappa B (NF-κB) was detected by Western blot and immunofluorescence. Results: Compared with the Control group and the ALDH2 NC group, the expression of ALDH2 was up-regulated in the ALDH2 group (P<0.05). Compared with the Control group and the siRNA-ALDH2 NC group, the expression of ALDH2 was down-regulated in the siRNA-ALDH2-1, -2, -3 groups (P<0.05), and the expression of ALDH2 was the lowest in the siRNA-ALDH2-2 group. Compared with Control group and ALDH2 NC group, cell adhesive ability, the expression of NF-κB in nucleus and the red fluorescence was decreased (P<0.05), and the expression of NF-κB in cytoplasm increased in ALDH2 group (P<0.05). Compared with Control group and siRNA-ALDH2 NC group, cell adhesive ability, the expression of NF-κB in nucleus and the red fluorescence was increased (P<0.05), the expression of NF-κB in cytoplasm decreased in siRNA-ALDH2 group (P<0.05). Conclusion: ALDH2 could inhibit the adhesion between THP-1-HAECs by inhibiting NF-κB nuclear transcription, and finally alleviate atherosclerosis.