基因过氧化物酶体增殖物激活受体γ共激因子-1α／雌激素相关受体-α共修饰间充质干细胞对血管化的影响

The effects of peroxisome - proliferator - activated receptor - γ coactivator - 1α and estrogen - related receptor - α gene - modified mesenchymal stem cells on angiogenesis in vitro

目的观察基因过氧化物酶体增殖物激活受体γ共激因子-1α（PGC-1α）／雌激素相关受体-α（ERR-α）共修饰间充质干细胞（MSCs）对血管化的影响。方法利用基因PGC-1α和ERR-α腺病毒过表达载体修饰MSCs；反转录-聚合酶链反应（RT-PCR）或酶联免疫吸附试验（ELISA）检测MSCs基因和促血管化细胞因子的变化；凝胶成血管实验观察MSCs对脐静脉血管内皮细胞（HUVECs）成血管能力的影响。结果基因PGC-1α/ERR—α共修饰可将细胞PGC-1α的mRNA表达水平提高至原表达的6倍；PGC-1α／ERR-α共修饰MSCs的血管内皮生长因子（VEGF）、血管生成素-2、血小板源生长因子-B（PDGF—B）mRNA表达显著高于未修饰和PGC-1α修饰MSCs（VEGF：5．45±0．51，1．00±0．24，2．33±0．31，F=189．100，P=0．000；ANGPT-2：2．66±0．38，1．00±0．29，1．89±0．24，F=36．180，P=0．000；PDGF—B：4．87±0．66，1．00±0．17，2．13±0．45，F=89．060，P=0．000），其培养上清VEGF分泌量显著高于未修饰和PGC-1α修饰MSCs（F=343．700，P=0．000）；与PGC-1α／ERR-α共修饰MSCs共培养HUVECs形成条索样血管结构的总长度[（82580±3156）μm]显著高于与未修饰[（9130±1174）μm]或PGC-1α修饰MSCs[（18570±2087）μm]的HUVECs（P=0．000）。结论基因PGC-1α/ERR-α共修饰可有效的加强MSCs的促血管化能力，有望用于组织工程血管化。

Objective To investigate the effects of mesenchymal stem ceils （MSCs） modified with peroxisome - proliferator - activated receptor - γ coactivator - 1α （ PGC - 1 α） and estrogen - related receptor - α （ ERR - ct） on angiogenesis in vitro. Methods The overexpression of PGC - 1 α and ERR - α was performed by an adenoviral vector encoding PGC - 1α and ERR - α. Proangiogenic factor expression of mesenchymal stem cells （MSCs） modified with genes were examined by Real -time polymerase chain reaction and enzyme - linked immunosorbent assay. The angiogenic capability of human umbilical vein endothelial ceils （ HUVECs）, which co - cultured with MSCs, was observed using a matrigel - based in vitro angiogenesis assay. Results Co - transfection of PGC - 1 α and ERR - α into MSCs led to a sixfold induction of PGC - 1α mRNA. Expression of proangiogenic factors such as vascular endothelial growth factor （ VEGF）, angiopoietin - 2 （ ANGPT - 2）, platelet - derived growth factor - B （ PDGF - B） were significantly increased in MSCs modified with PGC - 1α/ERR - α when compared with MSCs or PGC - 1 α - modified MSCs （VEGF： 5.45 ± 0.51, 1.00 ± 0.24, 2.33 ± 0.31, F= 189.100, P= 0.000; ANGPT-2： 2.66±0.38, 1.00±0.29, 1.89±0.24, F=36.180, P=0.000; PDGF- B： 4.87 ± 0. 66, 1.00 ± 0. 17, 2. 13 ± 0.45, F = 89. 060, P = 0. 000）. Levels of VEGF secreted by MSCs modified with PGC - 1α/ERR - α from culture supernatant were higher than that of MSCs modified with or without PGC - 1α （ F = 343. 700, P = 0. 000）. HUVECs cocultured with PGC - 1 α/ERR -α - modified MSCs resuited in a marked increase in the length of their capillary - like tube （ P = 0. 000）. Conclusion Our resuits demonstrated that the proangiogenic capability of MSCs was enhanced by modification with PGC - 1 α/ ERR - α for the application of vascularized tissue engineering.