论文部分内容阅读
目的探索血管内皮生长因子(VEGF)对间充质干细胞(MSC)增殖的影响及其信号机制。方法采用经典的全骨髓贴壁法培养MSC,通过成骨、成脂肪等多向诱导分化以及流式细胞仪分析其表面标记(CD45、CD34、CD90、CD29)等鉴定MSC特征;以P3MSC为实验材料,采用MTT法分析VEGF对MSC增殖的影响。随后以50 nmol/L Wortmannin、50μmol/LPD98059、30μmol/L SB203580、10μmol/L H89、20μmol/L Y27632、1μmol/L Rapamycin、10μmol/L Straurosporine、6nmol/L Gǒ6976、50μmol/L Pseudo Z等分别处理P3MSC,观察VEGF影响MSC增殖的信号机制。结果培养的P3MSC呈现出CD90、CD29强阳性,具有成骨、成脂肪等多向分化能力;VEGF促进MSC增殖,Y27632、PD98059、SB203580、Gǒ6976、Straurosporine可抑制VEGF促进MSC增殖的效应。结论 VEGF引起的MSC增殖效应与AKT-mTOR-PKC途径密切相关,且PKCα处于中间环节。
Objective To explore the effect of vascular endothelial growth factor (VEGF) on proliferation of mesenchymal stem cells (MSCs) and its signal mechanism. Methods MSCs were cultured by classical whole bone marrow adherent method. MSCs were identified by multi-directional induction of osteogenic and adipogenic differentiation and surface markers (CD45, CD34, CD90, CD29) by flow cytometry. Materials, the use of MTT assay of VEGF on the proliferation of MSC. Followed by 50 nmol / L Wortmannin, 50 μmol / LPD98059, 30 μmol / L SB203580, 10 μmol / L H89, 20 μmol / L Y27632, 1 μmol / L Rapamycin, 10 μmol / L Straurosporine, 6 nmol / L Gǒ6976, 50 μmol / L Pseudo Z, P3MSC, observe the signal mechanism of VEGF affecting MSC proliferation. Results The cultured P3MSCs showed strong positive of CD90 and CD29, with the ability of osteoblastic and adipogenic differentiation. VEGF promoted the proliferation of MSCs. Strains of Y27632, PD98059, SB203580, Gǒ6976 and Straurosporine could inhibit the proliferation of MSC. Conclusion The proliferation of MSC induced by VEGF is closely related to the AKT-mTOR-PKC pathway, and PKCα is in the middle part.