Welgus H G, Stricklin G P
J Biol Chem. 1983 Oct 25;258(20):12259-64.
In order to gain insight into the biological significance of a collagenase inhibitor secreted by human skin fibroblasts, we examined various human connective tissues and body fluids for such a protein. The inhibitors found in these tissues were compared immunologically to skin fibroblast inhibitor by Ouchterlony analysis and by the development of a highly specific enzyme-linked immunosorbent assay (ELISA). Using this ELISA, cell cultures of human skin fibroblasts, corneal fibroblasts, gingival fibroblasts, and adult and fetal lung fibroblasts secreted similar amounts of immunoreactive inhibitor protein. Each culture medium displayed a reaction of immunologic identity with skin fibroblast inhibitor when examined in Ouchterlony gel diffusion. In testing for functional inhibitory activity, the same 1:1 stoichiometry of collagenase inhibition was observed in each culture medium that characterizes the human skin inhibitor. Other mesodermally derived human cell types, including human fetal osteoblasts, uterine smooth muscle cells, fibrosarcoma cells, and explants of tendon and articular cartilage behaved in the same manner as the fibroblast cultures. Because collagenase inhibitors with biochemical similarities to skin fibroblast inhibitor have been described in serum and in amniotic fluid, we also examined these sources of inhibitory proteins. The data indicate that both serum and amniotic fluid contain collagenase inhibitors which are immunologically and functionally identical with the skin fibroblast inhibitor. The concentration of inhibitor in serum, as measured by ELISA assay, is 1.03 +/- 0.27 micrograms/ml. The results suggest that collagenase inhibitors which are functionally equivalent and immunologically identical with human skin fibroblast collagenase inhibitor are synthesized by many, if not all, fetal and adult mesodermal tissues in the human organism. The inhibitor apparently gains access to certain body fluids such as serum and amniotic fluid. This inhibitor protein may, therefore, function in the regulation of collagen degradation in most human connective tissues.
为深入了解人皮肤成纤维细胞分泌的胶原酶抑制剂的生物学意义，我们检测了各种人体结缔组织和体液中是否存在这种蛋白质。通过免疫双扩散分析以及开发一种高度特异性的酶联免疫吸附测定法（ELISA），将这些组织中发现的抑制剂与皮肤成纤维细胞抑制剂进行免疫比较。使用这种ELISA，人皮肤成纤维细胞、角膜成纤维细胞、牙龈成纤维细胞以及成人和胎儿肺成纤维细胞的细胞培养物分泌出相似量的免疫反应性抑制剂蛋白。当在免疫双扩散凝胶中检测时，每种培养基与皮肤成纤维细胞抑制剂都显示出免疫同一性反应。在测试功能抑制活性时，在每种培养基中都观察到了与人体皮肤抑制剂相同的1:1化学计量比的胶原酶抑制作用。其他中胚层来源的人体细胞类型，包括人胎儿成骨细胞、子宫平滑肌细胞、纤维肉瘤细胞以及肌腱和关节软骨外植体，其行为与成纤维细胞培养物相同。由于在血清和羊水中已描述了与皮肤成纤维细胞抑制剂具有生化相似性的胶原酶抑制剂，我们也检测了这些抑制蛋白的来源。数据表明，血清和羊水都含有与皮肤成纤维细胞抑制剂在免疫和功能上相同的胶原酶抑制剂。通过ELISA测定法测得血清中抑制剂的浓度为1.03±0.27微克/毫升。结果表明，在人体中，许多（如果不是全部）胎儿和成人中胚层组织都会合成与人体皮肤成纤维细胞胶原酶抑制剂在功能上等效且免疫上相同的胶原酶抑制剂。这种抑制剂显然能够进入某些体液，如血清和羊水。因此，这种抑制蛋白可能在大多数人体结缔组织中参与胶原降解的调节。
