【摘要】 [目的]观察SDF1/CXCR4对髓核细胞(NPCs)诱导血管内皮细胞(VECs)成血管能力的影响。[方法]通过病毒转染上调原代髓核细胞中基质细胞衍生因子1(SDF1)表达,并根据表达量分为D组和UP组,将不同分组的髓核细胞(或条件培养基)与血管内皮细胞进行共培养,分别进行细胞计数试剂盒8(CCK8)、细胞迁移实验、管腔形成实验,以观察髓核细胞对于血管内皮细胞成血管能力的影响。[结果]在mRNA和蛋白水平检验证实SDF1表达在UP组上调成功。NPCs和VECs共培养后,UP组VECs中pAKT水平较D组显著升高,PTEN水平较D组下降,差异均具有统计学意义。后续的共培养体系中,UP组的VECs在细胞活力、迁移能力、形成管腔样结构能力上较D组均显著增强,差异具有统计学意义。加入MK-2206后,UP+MK-2206组在细胞活力、迁移能力、管腔样结构形成能力上较UP组均显著下降,差异具有统计学意义。加入SF1670后,UP+SF1670组的细胞活力、迁移能力、管腔样结构形成能力均较UP组进一步增强,差异具有统计学意义。[结论]本实验通过体外培养证明退变髓核细胞可通过SDF1/CXCR4信号轴来诱导血管内皮细胞成血管活动,并且这一活动在内皮细胞内通过PI3K/AKT通路调控。更多还原

【Abstract】 [Objective] To investigate the influence of SDF1/CXCR4 axis on angiogenesis of vascular endothelial cells(VECs) induced by nucleus pulposus cells(NPCs). [Methods] The expression of stromal cell derived factor 1(SDF1) in primary nucleus pulposus cells was up-regulated by adenovirus transfection. The NPCs were classified as D(degenerated) and UP(upregulation) groups according to the expression of SDF1.NPCs or NPCs conditional medium from different groups were co-cultured with VECs, MK-2206, the inhibitor of phosphorylated AKT(pAKT), and SF1670, the inhibitor of phosphate and tension homology deleted on chromsome ten(PTEN), were added in the subsequent co-culture. Cell count kit 8(CCK8), cell migration test, tube formation test were used to observe the influence of NPCs on angiogenesis of VECs. [Results] Tests in mRNA and protein level documented that SDF1 expression in UP group was successfully up-regulated. After co-cultur of NPCs with VECs,pAKT of VECs in UP group was higher than D group, PTEN in UP group was lower than D group, and both of the results had statistical difference. In the subsequent co-culture system, cell viability, migration and tube formation ability of VECs in UP group were significantly elevated and had a significant difference compared with D group. After adding MK-2206, cell viability, migration and tube formation ability of VECs in UP + MK-2206 group were significantly declined and had a significant difference compared with UP group. After adding SF1670, cell viability, migration and tube formation ability of VECs in UP + SF1670 group were significantly enhanced and had a significant difference compared with UP group. [Conclusion] This in vitro experiment demonstrates that NPCs could induce angiogenesis of VECs though SDF1/CXCR4 axis, and this VECs activity is regulated internally by PI3 K/AKT passway.更多还原