Research Article| Volume 65, ISSUE 3, P217-225, September 2004

High glucose reduces albumin uptake in cultured proximal tubular cells (LLC-PK1)


      In this study, we clarify that high glucose inhibits albumin uptake in cultured LLC-PK1 cells. LLC-PK1 cells cultured for 6 days with 5.5–27.8 mM d-glucose were challenged by fluorescein isothiocyanate (FITC)-conjugated human albumin (HA). FITC-HA binding and uptake were inhibited by >5.5 mM glucose (Math Eq mM). Analysis of FITC-HA binding and uptake at 5.5 and 16.7 mM d-glucose (high glucose, HG) showed decreased affinity (Km for binding: 35.5 mg/l versus 52.6 mg/l, Km for uptake; 41.3 mg/l versus 55.6 mg/l) and maximal velocity (Bmax—0.33 μg versus 0.27 μg/30 min/mg protein; Umax—4.40 μg versus 3.48 μg/60 min/mg protein) at HG. A comparison of the time courses of FITC-HA binding and uptake at 5.5 mM glucose and at HG showed that HG suppressed them beyond 15 min (P<0.05–0.001). Phlorizin (>0.25 mM) completely reversed the HG-induced inhibition of FITC-HA binding and uptake. High glucose decreased mRNA of GLUT-1 and SGLT-1, but did not influence that of SGLT-2. The simultaneous presence of Vitamin E (10−6 M), Vitamin C (10−6 M) and reduced glutathione (0.25 mM) reversed the suppressed FITC-HA binding and uptake by HG, while any one or two of these molecules, and various inhibitors of advanced glycation end products, failed to do so.
      In conclusion, a high glucose milieu causes inhibition of albumin binding and uptake in proximal tubular cells by increasing metabolic oxidative stress through excessive glucose flux via the sodium glucose transporter.


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