5 years ago

Spontaneous PMN apoptosis in type 2 diabetes and the impact of periodontitis.

Stephens, Van Dyke, Kantarci, Manosudprasit, Hasturk
The purpose of this study was to test the hypothesis that peripheral blood neutrophils (PMN) exhibit delayed spontaneous apoptosis in individuals with diabetes mellitus type 2 (T2DM) and that the delay is exacerbated further among people who coexpress chronic periodontitis (CP). Seventy-three individuals were enrolled, including those with T2DM (n = 16), CP (n = 15), T2DM + CP (n = 21), and healthy volunteers (n = 21). PMN apoptosis was determined by flow cytometry using TUNEL and Annexin V assays. The activity of caspase-3, -8, and -9 was measured by colorimetric assay. PMN surface death receptor quantification was performed by flow cytometry staining with fluorescence-conjugated anti-CD120a (TNFR1) and anti-CD95 [Fas receptor (FasR)] antibody. Analysis of inflammatory markers in serum samples was performed using multiplexed sandwich immunoassays. In healthy volunteers and individuals with T2DM, CP, and T2DM + CP, spontaneous PMN apoptosis observed at 12 h reached 85.3 ± 3.1, 67.3 ± 3.9, 62.9 ± 3.5 and 62.5 ± 5.4%, respectively (P < 0.05). Caspase-3 activity was significantly reduced in individuals with T2DM and T2DM + CP (P < 0.05) when compared with healthy volunteers. Caspase-8 activity was also significantly decreased in CP and T2DM + CP (P < 0.05), associated with reduced cell-surface FasR, TNFRs, and Fas ligand (FasL) serum levels. Glucose alone was not observed to impact PMN apoptosis; simultaneous incubation with the receptor for advanced glycation endproducts (RAGE) agonist S100B induced significant PMN apoptosis (P < 0.05). These data support the premise that the inhibition of PMN apoptosis in individuals with T2DM occurs through an advanced glycation endproducts/RAGE ligand/receptor-mediated interaction.

Publisher URL: http://doi.org/10.1189/jlb.4A0416-209RR

DOI: 10.1189/jlb.4A0416-209RR

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