Moderately controlled transport of ascorbate into aortic endothelial cells against slowdown of the cell cycle, decreasing of the concentration or increasing of coexistent glucose as compared with dehydroascorbate.
Uptake of L-[1-(14)C]ascorbic acid (Asc) of 12.5-200 microM for 1 h into bovine aortic endothelial BAE-2 cells grown to confluence was as low as 43-64% (per cell) of uptake into the cells grown to nearly one-fourth confluence. [14C]Asc undergoing transmembrane uptake was concentrated and accumulated in the cell less efficiently ([Asc]in/ex = 8-13) at confluence than at subconfluence ([Asc]in/ex = 15-24). The declined Asc uptake at confluence is attributable to slowdown of the cell cycle, because a similar decrease in [Asc]in/ex was shown by subconfluent cells precultured in serum-insufficient medium, resulting in an increase in G1 phase and concurrent decreases in S and G2 + M phase distributions as determined by flow cytometry. [1-(14)C]Dehydroascorbic acid (DehAsc) was taken up and accumulated as Asc, after metabolic reduction, without detectable DehAsc. The [Asc]in/ex values for DehAsc at confluence were as low as 15-69% of those at subconfluence in contrast to the values as retentive as 62-75% for Asc, suggesting the moderate control of Asc uptake against slowdown of the cell cycle. At either confluence or subconfluence, dose-dependence for DehAsc uptake was more marked than for Asc uptake as shown by an uphill slope in a curve of doses versus [Asc]in/ex for DehAsc in contrast to a downhill slope for Asc, suggesting the moderate control for Asc uptake against fluctuation of the dose. Increasing of coexistent glucose of 5 mM to 20-40 mM, plasma concentrations in diabetic patients, declined DehAsc uptake to 46-48%, which was less moderately controlled than Asc uptake retained to 59-73%. Asc uptake did not compete with DehAsc uptake, suggesting different transporter proteins for Asc and DehAsc. Thus, Asc uptake into the aortic endothelial cells is more moderately controlled against slowdown of the cell cycle, decreasing of the extracellular concentrations or increasing of coexistent glucose than DehAsc uptake, suggesting a homeostatic advantage of Asc over DehAsc in terms of retention of intracellular Asc contents within a definite range.
Saitoh
Y
Y
Department of Cell Biochemistry, Hiroshima Prefectural University School of BioScience, Shobara, Japan.
Nagao
N
N
O'Uchida
R
R
Yamane
T
T
Kageyama
K
K
Muto
N
N
Miwa
N
N
D000818:Animals; D001011:Aorta / Q000166:cytology; D001205:Ascorbic Acid / Q000493:pharmacokinetics*; D001693:Biological Transport, Active / Q000187:drug effects / Q000502:physiology; D002250:Carbon Radioisotopes; D002417:Cattle; D002453:Cell Cycle / Q000502:physiology*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D003683:Dehydroascorbic Acid / Q000493:pharmacokinetics*; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology*; D005947:Glucose / Q000008:administration & dosage / Q000494:pharmacology*; D007424:Intracellular Fluid / Q000737:chemistry; D010084:Oxidation-Reduction
D002250:Carbon Radioisotopes; D016895:Culture Media, Serum-Free; D005947:Glucose; D001205:Ascorbic Acid; D003683:Dehydroascorbic Acid