【摘要】 目的 探讨机械敏感性离子通道Piezo1对血管内皮生长因子(VEGF)、血小板源性生长因子(PDGF)、转化生长因子-β(TGF-β)在静脉畸形(Venous malformations, VMs)细胞模型中促血管生成的作用。方法 通过慢病毒转染人脐静脉内皮细胞系(HUVEC),构建TIE2-L914F与TIE2-WT过表达的细胞模型，分别为L914F组和WT组，未转染的HUVEC细胞设为Control组。通过CCK-8实验检测YODA1(Piezo1的激动剂)和GsMTx4(Piezo1的抑制剂)药物对细胞毒性的影响。采用Western blot检测Piezo1、VEGF、PDGF和TGF-β蛋白的表达水平。结果 CCK-8结果显示，在0.1～6μmol/L YODA1浓度和0.5～3μmol/L GsMTx4浓度下，干预12 h各组细胞增殖作用能够持续，细胞存活率均在80%以上，最终确定后续实验YODA1干预浓度为6μmol/L,GsMTx4干预浓度为3μmol/L。Western blot结果显示，在WT组与L914F组中，Piezo1蛋白表达低于Control组(P<0.05),GsMTx4(3μmol/L)干预12 h后，PDGF、VEGF和TGF-β蛋白表达水平均较未干预组降低(P<0.05),YODA1(6μmol/L)干预12 h后，VEGF、PDGF和TGF-β蛋白表达水平均较未干预组升高(P<0.05)。结论 VMs细胞模型中Piezo1表达低于Control组，提示Piezo1参与VMs的发生，通过YODA1与GsMTx4的干预，提示Piezo1参与调控血管生长因子VEGF、PDGF和TGF-β的表达，从而促进VMs疾病的发展。更多还原

【Abstract】 Objective To investigate the role of mechanosensitive ion channel Piezo1 on the angiogenesis of vascular endothelial growth factor(VEGF), platelet-derived growth factor(PDGF), and transforming growth factor-β(TGF-β) in the venous malformation(VMs) cell model. Methods Construction of a cellular model of TIE2-L914F and TIE2-WT overexpression by lentiviral transfection of human umbilical vein endothelial cells(HUVEC), which were named L914F group and WT group respectively. UntransfectedHUVEC cells were set up as Control group.The cytotoxicity effects of YODA1(an agonist of Piezo1) and GsMTx4(an inhibitor of Piezo1) were detected by CCK-8 assay. The expression levels of Piezo1, VEGF, PDGF and TGF-β proteins were detected by Western blot. Results The CCK-8 assay results showed that 0.1-6 μmol/L concentration of YODA1 and 0.5-3 μmol/L concentration of GsMTx4, the proliferation of cells in each group could be sustained for 12 hours, with cell survival rates all above 80%. The final determination of the subsequent experiments was that the intervention concentration of YODA1 was 6 μmol/L, and the intervention concentration of GsMTx4 was 3 μmol/L.Western blot results showed that the protein expression levels of Piezo1 in L914F and WT groups were lower than that in Control group(P<0.05), and the protein expression levels of PDGF, VEGF, and TGF-β were lower than those in the non-intervention group after 12 h of GsMTx4(3 μmol/L) intervention(P<0.05), the levels of VEGF, PDGF and TGF-β proteins were higher than those in the non-intervention group after 12 h of YODA1(6 μmol/L) intervention(P<0.05). Conclusion The expression of Piezo1 in the VMs cell model is lower than that in Control group, suggesting that Piezo1 is involved in the occurrence of VMs, and by the intervention of YODA1 and GsMTx4, it is suggested that Piezo1 is involved in the regulation of the expression of vascular growth factors, VEGF, PDGF and TGF-β, which promotes the development of VMs disease.更多还原