Impaired neutrophil store-mediated calcium entry in Type 2 diabetes.
BACKGROUND
In Type 2 diabetes impaired neutrophil function leads to increased bacterial infection and cardiovascular disease. Many neutrophil functions depend on calcium signalling, which involves release of calcium from intracellular stores and subsequently translocation of stores via the cytoskeleton to the plasma membrane, causing store-mediated calcium entry (SMCE) into the cell. We hypothesized that in Type 2 diabetes there would be a defect in SMCE.


METHODS
Neutrophils were prepared from patients with Type 2 diabetes (DM, n=15) and controls (NC, n=15). Free cytosolic calcium [Ca2+]i was measured with Fura-2 in resting cells and after stimulation of calcium release with fMLP and thapsigargin.


RESULTS
Baseline [Ca2+]i was higher in neutrophils from the patients than the controls (NC 65 +/- 5 nm, DM 80 +/- 4 nm, P<0.05). However, after fMLP-treatment [Ca2+]i was significantly lower in the patients (NC 301 +/- 28 nm, DM 210 +/- 20 nm, P<0.01). The greater increase in controls was not observed when cells were treated with fMLP in the absence of extracellular calcium (-fold increase NC 2.9 +/- 0.5, DM 2.7 +/- 0.3). Treatment of cells with thapsigargin caused a similar greater increase in [Ca2+]i in the controls than in the patients that was not seen in the absence of extracellular calcium (-fold increase with Ca2+ NC 5.2 +/- 1.0, DM 3.0 +/- 0.4, P<0.05; fold increase without Ca2+ NC 2.5 +/- 0.4, DM 2.2 +/- 0.2).


CONCLUSIONS
In Type 2 diabetes there is a defect in neutrophil calcium signalling which results in a lesser increase in free cytosolic calcium owing to impaired influx across the plasma membrane. Abnormal calcium signalling is likely to be important in the pathogenesis of diabetic complications.
Department of Medicine, University of Newcastle Upon Tyne, Newcastle upon Tyne, UK. Andrew.Advani@ncl.ac.uk
A
A
Advani
S M
SM
Marshall
T H
TH
Thomas
D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology; D003600:Cytosol / Q000378:metabolism; D003924:Diabetes Mellitus, Type 2 / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D008875:Middle Aged; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D019284:Thapsigargin / Q000494:pharmacology
D004791:Enzyme Inhibitors; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019284:Thapsigargin; D002118:Calcium