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

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 m RNA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、siRNAALDH2 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。

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.