Effect of HUVEC on human osteoprogenitor cell differentiation needs heterotypic gap junction communication
Effect of HUVEC on human osteoprogenitor cell differentiation needs heterotypic gap junction communication
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DOI：
10.1152/ajpcell.00310.2001
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发表时间：
2002-04-01
影响因子：
5.5
通讯作者：
Amédée, J
中科院分区：
文献类型：
--
作者：
Villars, F;Guillotin, B;Amédée, J
骨发育和重塑依赖于骨形成成骨细胞和骨微环境中存在的其他细胞之间的复杂相互作用，特别是血管内皮细胞，其可能是骨中复杂的交互式通信网络的关键成员。本研究旨在探讨人脐静脉内皮细胞（HUVEC）与人骨髓基质细胞（HBMSC）之间的相互作用。不同细胞培养模型的细胞分化分析显示，碱性磷酸酶活性和I型胶原合成增加，只有HUVEC与HBMSC的直接接触。这种“阿曲他克林信号传导”可能涉及许多不同的异型连接，这些连接需要粘附分子或间隙连接。荧光黄染料偶联试验证明HUVEC和HBMSC之间存在功能性偶联。免疫细胞化学显示，连接蛋白43（Cx43），一个特定的间隙连接蛋白，不仅在HBMSC，但也在内皮细胞网络中表达，这两种细胞类型可以通过至少由Cx43构成的间隙连接通道进行通信。此外，通过18 α-大黄酸处理的差距连接的功能抑制或用寡脱氧核苷酸反义抑制Cx43合成降低了HUVEC共培养物对HBMSC分化的作用。这种刺激可以通过渗透连接通道的信号分子的细胞间扩散来介导。
Bone development and remodeling depend on complex interactions between bone-forming osteoblasts and other cells present within the bone microenvironment, particularly vascular endothelial cells that may be pivotal members of a complex interactive communication network in bone. Our aim was to investigate the interaction between human umbilical vein endothelial cells (HUVEC) and human bone marrow stromal cells (HBMSC). Cell differentiation analysis performed with different cell culture models revealed that alkaline phosphatase activity and type I collagen synthesis were increased only by the direct contact of HUVEC with HBMSC. This "juxtacrine signaling" could involve a number of different heterotypic connexions that require adhesion molecules or gap junctions. A dye coupling assay with Lucifer yellow demonstrated a functional coupling between HUVEC and HBMSC. Immunocytochemistry revealed that connexin43 (Cx43), a specific gap junction protein, is expressed not only in HBMSC but also in the endothelial cell network and that these two cell types can communicate via a gap junctional channel constituted at least by Cx43. Moreover, functional inhibition of the gap junction by 18alpha-glycyrrhetinic acid treatment or inhibition of Cx43 synthesis with oligodeoxyribonucleotide antisense decreased the effect of HUVEC cocultures on HBMSC differentiation. This stimulation could be mediated by the intercellular diffusion of signaling molecules that permeate the junctional channel.