[1]刘汝银,岳宗进,彭晓艳,等.血管内皮生长因子与转化生长因子的协同作用诱导兔骨髓间充质干细胞分化[J].第三军医大学学报,2016,38(15):1755-1761.
 Liu Ruyin,Yue Zongjin,Peng Xiaoyan,et al.Synergistic effect of VEGF and TGF-β1 in osteogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells[J].J Third Mil Med Univ,2016,38(15):1755-1761.
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血管内皮生长因子与转化生长因子的协同作用诱导兔骨髓间充质干细胞分化(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第15期
页码:
1755-1761
栏目:
基础医学
出版日期:
2016-08-15

文章信息/Info

Title:
Synergistic effect of VEGF and TGF-β1 in osteogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells
作者:
刘汝银岳宗进彭晓艳王新立冯仲锴
河南省中医院:脊柱科,护理部
Author(s):
Liu Ruyin Yue Zongjin Peng Xiaoyan Wang Xinli Feng Zhongkai

Department of Spine, Department of Nursing, Henan Provincial Hospital of Traditional Chinese Medicine, Zhengzhou, Henan Province, 450002, China

关键词:
血管内皮生长因子转化生长因子骨髓间充质干细胞椎间盘退变
Keywords:
vascular endothelial growth factor transforming growth factor-&beta1 bone marrow-derived mesenchymal stem cells degeneration of intervertebral disc
分类号:
R329.24; R331.22; R341
文献标志码:
A
摘要:

目的      研究血管内皮生长因子(vascular endothelial growth factor,VEGF)和转化生长因子(transforming growth factor-β1,TGF-β1)的协同作用对兔骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSCs)成骨分化的影响及其可能机制。      方法      将细胞分为4组:对照组、腺相关病毒(adeno-associated virus,AAV)-VEGF组、AAV-TGF-β1组和VEGF+TGF-β1组。穿刺法抽取兔骨髓液,分离培养原代BMSCs,分别构建AAV-VEGF和AAV-TGF-β1腺病毒载体转染BMSCs,Western blot检测各组细胞中VEGF、TGF-β1的蛋白表达。流式细胞术检测细胞凋亡情况,CCK8法检测细胞增殖能力,Western blot检测髓核细胞标志SOX-9、Ⅱ型胶原和蛋白聚糖的表达,以及P38MAPK信号通路相关蛋白P38、MAPKAPK2和HSP27的表达。加入P38MAPK的特异阻滞剂SB203580预处理BMSCs,检测VEGF和TGF-β1对BMSCs增殖、凋亡、分化及相关蛋白表达的影响。      结果      VEGF和TGF-β1可通过调节P38MAPK信号通路抑制BMSCs凋亡,促进BMSCs增殖,并向类髓核细胞分化,且在其协同作用下效果显著。抑制P38MAPK信号通路可反转VEGF和TGF-β1对BMSCs增殖分化能力的促进作用。      结论      VEGF和TGF-β1的协同作用能增强BMSCs增殖和分化的能力,提高胶原蛋白和蛋白聚糖等细胞外基质的表达,从而促进退变椎间盘的修复和再生。

Abstract:

Objective      To investigate the synergistic effect of vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) on osteogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells (BMSCs) and its possible mechanisms.       Methods      Cells were divided into 4 groups, i.e. control, adeno-associated virus (AAV)-VEGF group, AAV-TGF-β1 group and VEGF+TGF-β1 group. The bone marrow was extracted from rabbit femoral trochanter by puncturing and BMSCs were isolated. Primary-cultured BMSCs were transfected with AAV-VEGF and AAV-TGF-β1 adenoviral vectors, respectively. The protein levels of VEGF and TGF-β1 were measured by Western blotting. Cell apoptosis was detected by flow cytometry and cell proliferation was measured by CCK-8 assay. The expression of nucleus pulposus cell marker proteins (SOX-9, Collagen Ⅱ and Aggrecan) and P38MAPK signaling pathway-related proteins (P38, MAPKAPK and HSP27) were measured by Western blotting. BMSCs were pretreated with SB203580, a specific P38MAPK inhibitor. The effects of VEGF and TGF-β1 on the proliferation, apoptosis, differentiation of treated BMSCs and associated protein expressions were measured.       Results      VEGF or TGF-β1 inhibited BMSCs apoptosis, increased BMSCs proliferation and promoted their differentiation to nucleus pulposus-like cells through P38MAPK signaling pathway. Moreover, these effects were reinforced by synergistic action of VEGF and TGF-β1. Inhibiting P38MAPK pathway reversed the accelerating effect of VEGF and TGF-β1 on the proliferation and differentiation of BMSCs.       Conclusion      VEGF and TGF-β1 synergistically enhance BMSCs proliferation and differentiation, elevate the expression of extracellular matrix including collagen and proteoglycan, and thus promote the repair and regeneration of degenerative intervertebral discs.

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更新日期/Last Update: 2016-07-21