Shapiro D L, Livingston J N, Maniscalco W M, Finkelstein J N
Biochim Biophys Acta. 1986 Feb 21;885(2):216-20. doi: 10.1016/0167-4889(86)90091-1.
Type II alveolar epithelial cells (pneumocytes) were isolated to purity from adult rabbits and analyzed for the presence of cell surface insulin receptors and for effects of insulin on cells. Assays were performed on cells cultured for 24 h in Eagle's minimum essential medium. Insulin binding to cells in culture approached a steady-state level by 180 min at 15 degrees C and remained constant for at least 1 h. Competition experiments using native insulin, proinsulin and desoctapeptide supported specificity of binding. Scatchard analysis of binding revealed a class of high-affinity receptors with Kd = 1.5 X 10(-10) M and a low-affinity component with Kd = 4 X 10(-9) M. The number of receptors was estimated at 2000-4000/cell. Insulin added to cell cultures of type II pneumocytes in concentrations from 5 X 10(-11) to 5 X 10(-8) M resulted in a dose-related increase in uptake of 2-deoxyglucose by cells. Insulin also stimulated the incorporation of choline and glucose into phosphatidylcholine and disaturated phosphatidylcholine.
从成年兔中分离出II型肺泡上皮细胞(肺细胞)并纯化,分析细胞表面胰岛素受体的存在情况以及胰岛素对细胞的作用。在伊格尔最低必需培养基中培养24小时的细胞上进行测定。在15℃下,胰岛素与培养细胞的结合在180分钟时接近稳态水平,并至少保持1小时恒定。使用天然胰岛素、胰岛素原和去八肽的竞争实验支持结合的特异性。结合的Scatchard分析显示一类高亲和力受体,Kd = 1.5×10⁻¹⁰ M,以及一个低亲和力组分,Kd = 4×10⁻⁹ M。受体数量估计为2000 - 4000个/细胞。以5×10⁻¹¹至5×10⁻⁸ M的浓度添加到II型肺细胞培养物中的胰岛素导致细胞对2-脱氧葡萄糖摄取的剂量相关增加。胰岛素还刺激胆碱和葡萄糖掺入磷脂酰胆碱和二饱和磷脂酰胆碱中。