南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (04): 499-505.doi: 10.12122/j.issn.1673-4254.2020.04.08

• • 上一篇    下一篇

奥美沙坦酯通过上调 miR-3133 抑制衰老相关血管平滑肌细胞 的迁移和侵袭

张 艺，盛 帅，梁庆阳，张 莉   

  • 出版日期:2020-04-30 发布日期:2020-04-20
  • 基金资助:

Olmesartan inhibits age-associated migration and invasion of human aortic vascular smooth muscle cells by upregulating miR-3133 axis

  

  • Online:2020-04-30 Published:2020-04-20

HTML

PDF

900

摘要： 目的 探讨奥美沙坦酯（OMST）抑制衰老相关人主动脉血管平滑肌细胞（HA-VSMCs）迁移和侵袭能力的作用及微小RNA（miRNA）参与的机制。方法 HA-VSMCs体外培养，分别经博莱霉素（BLM）诱导衰老、OMST干预及瞬时转染miR-3133 NC/miR-3133 inhibitor。分别设置对照组：HA-VSMCs 正常培养；BLM 组：HA-VSMCs+BLM；OMST 组：HA-VSMCs+BLM+OMST；OMST+miR-3133 NC组：HA-VSMCs转染miR-3133 NC+BLM+OMST；OMST+miR-3133 inhibitor组：HA-VSMCs转 染miR-3133 inhibitor+BLM+OMST。划痕实验和Boyden小室侵袭实验分别测定HA-VSMCs迁移和侵袭力。明胶酶谱分析测定HA-VSMCs中金属基质蛋白酶-2（MMP-2）分泌。MiRNA生物芯片测定差异表达miRNA并行qRT-PCR验证。结果 与对照组相比，BLM组划痕实验和侵袭实验显示细胞迁移和侵袭力增加（[ 78.43±12.76）% vs（42.47±7.22）%；33.33±5.51 vs 13.00± 4.36，P<0.05]，MMP-2分泌水平增加[1.66±0.27 vs 0.87±0.13，P<0.05]；而OMST干预可抑制增加的细胞迁移/侵袭力和MMP-2分泌。生物芯片检测显示，以上3组存在差异miRNAs表达。qRT-PCR证实BLM组miR-3133表达下调，而OMST组miR-3133表达上调。与OMST组相比，OMST+miR-3133 inhibitor组HA-VSMC迁移和侵袭能力增加（[ 85.87±7.39）% vs（49.77±3.05）%；34.67±2.31 vs 20.00±4.58，P<0.05]；MMP-2分泌水平也增加（1.76±0.19 vs 0.94±0.10，P<0.05）。结论 OMST能抑制衰老HAVSMC的迁移和侵袭能力，这一过程可能是通过上调miR-3133表达实现的。

Abstract: Objective To explore the effects of olmesartan on age-associated migration and invasion capacities and microRNA (miRAN) axis in human aortic vascular smooth muscle cells (HA-VSMCs). Methods Cultured HA-VSMCs were divided into control group, bleomycin-mediated senescence (BLM) group and bleomycin + olmesartan treatment group. Wound-healing assay and Boyden chambers invasion assay were used to assess the changes in migration and invasion of the cells, gelatin zymography was used to analyze matrix metalloproteinase-2 (MMP-2) activation in the cells. The differentially expressed miRNAs were identified by miRNA microarray assay and validated by quantitative real-time PCR. MiR-3133 inhibitor was used to examine the effects of molecular manipulation of olmesartan on age-associated migration and invasion and MMP-2 activation in the cells. Results Compared with those of the control group, the percentage of the repopulated cells and the number of cells crossing the basement membrane increased significantly in BLM group [(78.43±12.76)% vs (42.47±7.22)%, P< 0.05; 33.33±5.51 vs 13.00±4.36, P<0.05]. A significant increase of MMP-2 activation was found in BLM group as compared with the control group (1.66 ± 0.27 vs 0.87 ± 0.13, P<0.05). Olmesartan significantly inhibited BLM-induced enhancement of cell migration and invasion and MMP-2 secretion in the cells. MiR-3133 was significantly downregulated in BLM group and upregulated in olmesartan group. Transfection with miR-3133 inhibitor significantly reversed the effects of olmesartan on age-associated migration and invasion of the cells [(85.87±7.39)% vs (49.77±3.05)%; 34.67±2.31 vs 20.00±4.58, P<0.05] and MMP-2 activation in the cells (1.76 ± 0.19 vs 0.94 ± 0.10, P<0.05). Conclusion Olmesartan inhibits the migration and invasion of ageassociated HA-VSMCs probably by upregulating of the miR-3133 axis.