Majack R A, Bornstein P
J Cell Biol. 1985 Feb;100(2):613-9. doi: 10.1083/jcb.100.2.613.
The effect of heparin on the biosynthetic phenotype of rat vascular smooth muscle cells (SMC) was investigated in vitro. Addition of heparin to the culture medium of early passage rat SMC resulted in a marked (3-15-fold) increase of a cell layer-associated Mr 60,000 protein that was sensitive to digestion by purified bacterial collagenase and contained significant amounts of hydroxyproline. Pulse-chase analysis of heparin-treated SMC revealed that the Mr 60,000 collagen was a primary and abundant product of these cells and was not processed extracellularly to a smaller form. The inductive effect of heparin could be mimicked by iota carrageenan or dextran sulfates but not by hyaluronic acid, dermatan sulfate, or chondroitin sulfates. The induction was concentration dependent with a maximal effect observed at a heparin concentration of 10 micrograms/ml. Synthesis of the Mr 60,000 collagen increased 18-24 h after addition of heparin to the cultures. Following induction and subsequent removal of heparin, synthesis of the protein remained maximal for at least 12 h and required 72 h to return to a basal level. These data demonstrate that the biosynthetic phenotype of vascular SMC in vitro can be controlled, at least in part, by heparin and related polyanions and suggest a role for similar molecules endogenous to the vessel wall in the regulation of SMC function.
在体外研究了肝素对大鼠血管平滑肌细胞(SMC)生物合成表型的影响。向早期传代大鼠SMC的培养基中添加肝素,导致与细胞层相关的分子量为60,000的蛋白质显著增加(3至15倍),该蛋白质对纯化的细菌胶原酶消化敏感,并含有大量羟脯氨酸。对肝素处理的SMC进行脉冲追踪分析表明,分子量为60,000的胶原蛋白是这些细胞的主要且丰富的产物,不会在细胞外加工成更小的形式。肝素的诱导作用可被ι-角叉菜胶或硫酸葡聚糖模拟,但不能被透明质酸、硫酸皮肤素或硫酸软骨素模拟。诱导作用呈浓度依赖性,在肝素浓度为10微克/毫升时观察到最大效应。向培养物中添加肝素后18至24小时,分子量为60,000的胶原蛋白合成增加。诱导并随后去除肝素后,该蛋白质的合成至少在12小时内保持最大值,需要72小时才能恢复到基础水平。这些数据表明,体外血管SMC的生物合成表型至少部分受肝素和相关聚阴离子控制,并提示血管壁内源性类似分子在SMC功能调节中起作用。