Holderbaum D, Ehrhart L A
J Cell Physiol. 1986 Feb;126(2):216-24. doi: 10.1002/jcp.1041260210.
Collagen, fibronectin, and nonfibrous protein biosynthesis were examined in cultures of rabbit arterial smooth muscle cells grown on tissue culture plastic precoated either with rabbit plasma fibronectin or bovine serum albumin. Cells seeded into fibronectin-coated wells appeared to reach confluence more quickly than counterparts grown on albumin-coated surfaces. Measurement 3H-thymidine incorporation into DNA by these cultures suggested that this was probably a consequence of more rapid and efficient cell attachment rather than an increased rate of proliferation of smooth muscle cells grown on fibronectin. In preconfluent cultures, the rates of collagen and fibronectin biosynthesis were reduced to 34 and 57%, respectively, on a per-cell basis in cultures grown on fibronectin-coated surfaces compared with cells grown on albumin-coated plasticware. In preconfluent cultures grown on fibronectin-coated surfaces, a greater percentage of the total fibronectin synthesized was incorporated into the cell layer. The distribution of newly synthesized collagen between culture medium and cell layer, however, was not affected by alteration of substratum composition. There was no difference in the rate of synthesis of noncollagen proteins between the two groups of preconfluent cells. In postconfluent cultures the rates of collagen and fibronectin biosynthesis were equivalent in both albumin- and fibronectin-treated cultureware. In preconfluent cultures, analyses of procollagens showed that the overall amounts of both types I and III procollagens were reduced in fibronectin-treated wells, indicating the reduction in collagen synthesis to be general and not type-specific. Although type V procollagen biosynthesis was not detected in either preconfluent group, it was found in postconfluent cultures. The reduction of fibronectin synthesis in cells grown in fibronectin-coated wells was significant as early as 4 hours after plating. Together, these findings suggest that cultured arterial smooth muscle cells are capable of deriving information from their substratum and regulating the biosynthetic rates of extracellular matrix components in response to the immediate needs of the cell.
在预先包被有兔血浆纤连蛋白或牛血清白蛋白的组织培养塑料上生长的兔动脉平滑肌细胞培养物中,对胶原蛋白、纤连蛋白和非纤维蛋白的生物合成进行了检测。接种到纤连蛋白包被孔中的细胞似乎比在白蛋白包被表面上生长的细胞更快达到汇合状态。对这些培养物中3H-胸腺嘧啶掺入DNA的测量表明,这可能是细胞更快、更有效地附着的结果,而不是在纤连蛋白上生长的平滑肌细胞增殖速率增加。在预汇合培养物中,与在白蛋白包被塑料制品上生长的细胞相比,在纤连蛋白包被表面上生长的培养物中,按每个细胞计算,胶原蛋白和纤连蛋白的生物合成速率分别降至34%和57%。在纤连蛋白包被表面上生长的预汇合培养物中,合成的总纤连蛋白中有更大比例被掺入细胞层。然而,新合成的胶原蛋白在培养基和细胞层之间的分布不受基质成分改变的影响。两组预汇合细胞之间非胶原蛋白的合成速率没有差异。在汇合后培养物中,白蛋白处理和纤连蛋白处理的培养器皿中胶原蛋白和纤连蛋白的生物合成速率相当。在预汇合培养物中,对前胶原蛋白的分析表明,在纤连蛋白处理的孔中,I型和III型前胶原蛋白的总量均减少,表明胶原蛋白合成的减少是普遍的,而非类型特异性的。尽管在任何一个预汇合组中均未检测到V型前胶原蛋白的生物合成,但在汇合后培养物中发现了它。早在接种后4小时,在纤连蛋白包被孔中生长的细胞中纤连蛋白合成的减少就很明显。总之,这些发现表明,培养的动脉平滑肌细胞能够从其基质中获取信息,并根据细胞的即时需求调节细胞外基质成分的生物合成速率。