Navab M, Liao F, Hough G P, Ross L A, Van Lenten B J, Rajavashisth T B, Lusis A J, Laks H, Drinkwater D C, Fogelman A M
Department of Medicine, School of Medicine, University of California, Los Angeles 90024-1679.
J Clin Invest. 1991 May;87(5):1763-72. doi: 10.1172/JCI115195.
Medium from cocultures of human aortic endothelial cells (HAEC) and smooth muscle cells (HASMC) taken from the same donor contained approximately two- to fourfold more macrophage colony-stimulating factor, granulocyte/macrophage colony-stimulating factor, and up to 5.1-fold more transforming growth factor beta than could be accounted for by the sum of the activities of media from equivalent numbers of HAEC and HASMC cultured separately. After pulse labeling, immunoprecipitated [35S]fibronectin and [14C]collagen were also found to be substantially increased in the coculture compared to the sum of HAEC and HASMC cultured separately. The cocultivation of HAEC and HASMC resulted in a 2.7-fold increase in connexin43 messenger RNA. When direct physical contact between HAEC and HASMC was prevented by a membrane that was permeable to medium, the levels of [35S]fibronectin and [14C]collagen in the coculture were significantly reduced. Monocytes cultured alone contained low levels of [35S]fibronectin and [14C]collagen but when added to the coculture there was up to a 22-fold increase in [35S]fibronectin and a 1.9-fold increase in [14C]collagen compared to the coculture alone. The increase in fibronectin was prevented in the presence of neutralizing antibody to interleukin 1 and antibody to interleukin 6 by 45% and 67%, respectively. Addition of monocytes to cocultures also induced the levels of mRNA for connexin43 by 2.8-fold. We conclude that the interaction of HAEC, HASMC, and monocytes in coculture can result in marked increases in the levels of several biologically important molecules and that increased gap junction formation between the cells and interleukins 1 and 6 may be partially responsible for these changes.
取自同一供体的人主动脉内皮细胞（HAEC）和平滑肌细胞（HASMC）共培养的培养基中，巨噬细胞集落刺激因子、粒细胞/巨噬细胞集落刺激因子的含量比等量单独培养的HAEC和HASMC培养基活性总和高约两到四倍，转化生长因子β的含量则高达5.1倍。脉冲标记后发现，与单独培养的HAEC和HASMC总和相比，共培养中免疫沉淀的[35S]纤连蛋白和[14C]胶原蛋白也显著增加。HAEC和HASMC共培养使连接蛋白43信使核糖核酸增加了2.7倍。当用对培养基可渗透的膜阻止HAEC和HASMC之间的直接物理接触时，共培养中[35S]纤连蛋白和[14C]胶原蛋白的水平显著降低。单独培养的单核细胞含低水平的[35S]纤连蛋白和[14C]胶原蛋白，但加入共培养体系后，与单独的共培养相比，[35S]纤连蛋白增加了高达22倍，[14C]胶原蛋白增加了1.9倍。在存在白细胞介素1中和抗体和白细胞介素6抗体的情况下，纤连蛋白的增加分别被阻止了45%和67%。向共培养体系中添加单核细胞还使连接蛋白43的信使核糖核酸水平提高了2.8倍。我们得出结论，共培养中HAEC、HASMC和单核细胞之间的相互作用可导致几种生物学上重要分子的水平显著增加，细胞间缝隙连接形成增加以及白细胞介素1和6可能部分导致了这些变化。
