Di Girolamo N, Underwood A, McCluskey P J, Wakefield D
Laboratory of Ocular Immunology, School of Pathology, University of New South Wales, Australia.
Diabetes. 1993 Nov;42(11):1606-13. doi: 10.2337/diab.42.11.1606.
Decreased wound healing and increased infection are major problems in patients with diabetes mellitus. Fibronectin plays a fundamental role in wound healing and acts as an opsonin for the phagocytosis of foreign antigens. The aim of this study was to ascertain the functional activity of plasma fibronectin from patients with diabetes mellitus. Initially, a modified Boyden chamber technique was used to measure cell migration on fibronectin purified from patient's plasma and an enzyme-linked immunosorbent assay was used to measure the binding of gelatin. A sandwich assay was developed that enabled the capture of fibronectin directly from patient's plasma without prior purification. With the use of a 96-well format, the binding of two different monoclonal antibodies could be compared simultaneously with the binding of gelatin and cell adhesion. In this way, differences in the function of particular domains of fibronectin from diabetic patients and control subjects could be measured. Results showed no difference between fibronectin from diabetic patients and control subjects with respect to the monoclonal antibodies binding in 1) the cell adhesion domain and 2) the heparin-binding domain. Furthermore, no detectable differences were noted with respect to cell adhesion, cell migration, or gelatin binding. These results suggest that diabetic patients receiving insulin treatment show no modulation of plasma fibronectin function, despite raised levels of circulating glucose.
伤口愈合减慢和感染增加是糖尿病患者的主要问题。纤连蛋白在伤口愈合中起重要作用，并作为一种调理素参与对外来抗原的吞噬作用。本研究的目的是确定糖尿病患者血浆纤连蛋白的功能活性。最初，采用改良的博伊登小室技术来测量细胞在从患者血浆中纯化的纤连蛋白上的迁移，并使用酶联免疫吸附测定法来测量明胶的结合。开发了一种夹心测定法，可直接从患者血浆中捕获纤连蛋白而无需事先纯化。使用96孔板形式，可以同时比较两种不同单克隆抗体的结合与明胶结合及细胞黏附的情况。通过这种方式，可以测量糖尿病患者和对照受试者纤连蛋白特定结构域功能的差异。结果显示，糖尿病患者的纤连蛋白与对照受试者的纤连蛋白在以下方面无差异：1）细胞黏附结构域和2）肝素结合结构域的单克隆抗体结合。此外，在细胞黏附、细胞迁移或明胶结合方面未观察到可检测到的差异。这些结果表明，接受胰岛素治疗的糖尿病患者尽管循环葡萄糖水平升高，但血浆纤连蛋白功能未发生调节。
