摘要目的 探讨miR-34b-3p对衰老血管内皮细胞的增殖、迁移和管腔形成功能的调控作用.方法 体外培养原代人脐静脉内皮细胞(HUVECs),通过传代计算细胞群体倍增水平(PDLs),建立年轻内皮细胞模型(PDL8)和衰老内皮细胞模型(PDL44).应用逆转录实时荧光定量聚合酶链反应(RT-qPCR)检测miR-34b-3p在PDL8和PDL44 HUVECs中的表达情况.分别在PDL8 HUVECs、PDL44 HUVECs中过表达和抑制miR-34b-3p,采用细胞增殖-毒性检测试剂盒-8(CCK-8)、迁移和管腔形成实验检测HUVECs的增殖、迁移和管腔形成功能.结果 miR-34b-3p在PDL44 HUVECs中的表达量较PDL8 HUVECs中的表达量上调约4.3倍,差异有统计学意义(t=-4.528,P＜0.05).PDL8HUVECs空白转染组内皮细胞的增殖率、迁移率、管腔总长度和管腔节点数分别较PDL44 HUVECs空白转染组升高1.2倍(0.67/0.57)、1.2倍(106/86)、1.4倍(10 605/7 735)和1.3倍(41/31),差异均有统计学意义(t=3.237、3.564、5.165、3.487,P＜0.05或P＜0.01);在PDL8 HUVECs中过表达miR-34b-3p可以使内皮细胞的增殖率、迁移率、管腔总长度和管腔节点数分别降低2.2倍(0.67/0.30)、2.3倍(106/46)、1.6倍(10 605/6 652)和1.9倍(41/22),差异均有统计学意义(F=145.898、53.026、41.997、36.341,均P＜0.01);在PDL44 HUVECs中抑制miR-34b-3p可以使内皮细胞增殖率、迁移率、管腔总长度和管腔节点数分别升高1.4倍(0.77/0.57)、2.3倍(198/86)、1.7倍(13 073/7 735)和2.3倍(71/31),差异均有统计学意义(F=14.815、42.970、167.063、258.340,均P＜0.01).结论 miR-34b-3p在衰老HUVECs中的高表达可以导致衰老血管内皮细胞增殖、迁移和管腔形成功能障碍.

abstractsObjective To investigate the effect of miR-34b-3p on the proliferation,migration and tube formation of senescent endothelial cell.Methods Primary human umbilical vein endothelial cells (HUVECs) were cultured in vitro,and population doubling levels (PDLs) were calculated by passage.The young endothelial cell was defined as PDL8.The senescent endothelial cell was defined as PDL44.Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was applied to detect the expression of miR-34b-3p in PDL8 and PDL44 HUVECs.miR-34b-3p mimic and inhibitor were transfected into PDL8 and PDL44 HUVECs.Then,cell counting kit-8 (CCK-8),transwell and tube formation assays were used to determine the proliferation,migration and tube formation of HUVECs,respectively.Results miR-34b-3p was significantly up-regulated approximately 4.3 times in PDL44 HUVECs than that in PDL44 HUVECs (t =-4.528,P ＜ 0.05).The proliferation,migration,total tube length and branch points of miR-34b-3p in PDL8 HUVECs group were significantly higher approximately 1.2 (0.67/0.57),1.2 (106/86),1.4 (10 605/7 735) and 1.3 (41/31) times than that in PDL44 HUVECs group,respectively (t =3.237,3.564,5.165,3.487,P ＜ 0.05 or P ＜ 0.01).Overexpression of miR-34b-3p had significantly inhibited proliferation,migration,total tube length and branch points approximately 2.2 (0.67/0.30),2.3(106/46),1.6 (10 605/6 652) and 1.9 (41/22) times in PDL8 HUVECs,respectively (F=145.898,53.026,41.997,36.341,all P ＜ 0.01).Repression of miR-34b-3p had significantly increase proliferation,migration,total tube length and branch points approximately 1.4 (0.77/0.57),2.3 (198/86),1.7 (13 073/7 735) and 2.3 (71/31) times in PDL44 HUVECs,respectively (F =14.815,42.970,167.063,258.340,all P ＜0.01).Conclusion The high expression of miR-34b-3p in senescent HUVECs could impair the proliferation,migration and tube formation of senescent endothelial cell.