Two pathways for insulin metabolism in adipocytes
Two pathways for insulin metabolism in adipocytes
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DOI：
10.1016/s0167-4889(97)00066-9
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发表时间：
1997-09-11
影响因子：
5.1
通讯作者：
Peavy, DE
中科院分区：
文献类型：
--
作者：
Duckworth, WC;Hamel, FG;Peavy, DE
使用选定的条件、适当的胶原酶、白蛋白和细胞处理，开发出不具有细胞外胰岛素降解活性的分离脂肪细胞制剂。使用三氯乙酸沉淀性作为测量值，细胞介导的胰岛素降解率为 0.68% +/- 0.05%/100000 个细胞/小时。氯喹 (CQ) 增加细胞相关放射性并减少降解，而丹磺酰尸胺 (DC)、PCMBS 和杆菌肽 (BAC) 减少降解，但对结合没有影响。细胞相关放射性的提取和色谱显示3个峰，一个大分子量峰、一个小分子量峰和一个胰岛素大小的峰。 CQ、DC和BAC均降低了小分子量峰，而CQ和DC还增加了大分子量放射性峰。氯喹是胰岛素降解酶 (IDE) 的细胞介导抑制剂（杆菌肽和 PCMBS），另一种是通过细胞加工抑制剂（DC、CQ 和氧化苯砷）改变的，氯喹改变了胰岛素大小的细胞相关 HPLC 测定的降解产物的模式，进一步支持了两种降解途径；一种是氯喹敏感途径，一种是氯喹不敏感途径。 (C) 1997 Elsevier Science B.V.
Using selected conditions, the appropriate collagenase, albumin and cell treatment, a preparation of isolated adipocytes was developed with no extracellular insulin degrading activity. Cell mediated insulin degradation rates were 0.68% +/- 0.05%/100000 cell/h using trichloracetic acid precipitability as a measure. Chloroquine (CQ) increased cell-associated radioactivity and decreased degradation while dansylcadaverine (DC), PCMBS and bacitracin (BAC) decreased degradation with no effect on binding. Extraction and chromatography of the cell-associated radioactivity showed 3 peaks, a large molecular weight peak, a small molecular weight peak and an insulin-sized peak. CQ, DC and BAC all decreased the small molecular weight peak while CQ and DC also increased the peak of large molecular weight radioactivity. Cell mediated inhibitors of the insulin degrading enzyme (IDE) (bacitracin and PCMBS) and the other altered by cell processing inhibitors (DC, CQ and phenylarsenoxide), Chloroquine altered the pattern of the insulin-sized cell-associated HPLC assayed degradation products, further supporting two pathways of degradation; one a chloroquine-sensitive and one a chloroquine-insensitive pathway. (C) 1997 Elsevier Science B.V.