Effect of insulin and glucose on adenosine metabolizing enzymes in human B lymphocytes.
In diabetes several aspects of immunity are altered, including the immunomodulatory action of adenosine. Our study was undertaken to investigate the effect of different glucose and insulin concentrations on activities of adenosine metabolizing enzymes in human B lymphocytes line SKW 6.4. The activity of adenosine deaminase in the cytosolic fraction was very low and was not affected by different glucose concentration, but in the membrane fraction of cells cultured with 25 mM glucose it was decreased by about 35% comparing to the activity in cells maintained in 5 mM glucose, irrespective of insulin concentration. The activities of 5'-nucleotidase (5'-NT) and ecto-5'-NT in SKW 6.4 cells depended on insulin concentration, but not on glucose. Cells cultured with 10(-8) M insulin displayed an about 60% lower activity of cytosolic 5'-NT comparing to cells maintained at 10(-11) M insulin. The activity of ecto-5'-NT was decreased by about 70% in cells cultured with 10(-8) M insulin comparing to cells grown in 10(-11) M insulin. Neither insulin nor glucose had an effect on adenosine kinase (AK) activity in SKW 6.4 cells or in human B cells isolated from peripheral blood. The extracellular level of adenosine and inosine during accelerated catabolism of cellular ATP depended on glucose, but not on insulin concentration. Concluding, our study demonstrates that glucose and insulin differentially affect the activities of adenosine metabolizing enzymes in human B lymphocytes, but changes in those activities do not correlate with the adenosine level in cell media during accelerated ATP catabolism, implying that nucleoside transport is the primary factor determining the extracellular level of adenosine.
Kocbuch
Katarzyna
K
Department of Molecular Medicine, Medical University of Gdansk, Debinki 7, paw. 29, Gdańsk, Poland. katkocbuch@gumed.edu.pl
Sakowicz-Burkiewicz
Monika
M
Grden
Marzena
M
Szutowicz
Andrzej
A
Pawelczyk
Tadeusz
T
D015720:5'-Nucleotidase / Q000378:metabolism; D000241:Adenosine / Q000378:metabolism*; D000243:Adenosine Deaminase / Q000378:metabolism; D000248:Adenosine Kinase / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D001402:B-Lymphocytes / Q000187:drug effects* / Q000201:enzymology*; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D009708:Nucleotidases / Q000378:metabolism
D007328:Insulin; D000255:Adenosine Triphosphate; D000248:Adenosine Kinase; D009708:Nucleotidases; C030287:nucleotidase; D015720:5'-Nucleotidase; D000243:Adenosine Deaminase; D005947:Glucose; D000241:Adenosine