Azize SENER, Nazli Gul ALTINDIS, Ozge DOGAN

Akut Hipergliseminin Trombositlerde Oksidatif Strese ve Nitrik Oksit Biyoyaralanımına Etkisi

Amaç: Diyabette, trombositlerin aktivasyona ve trombus oluşumuna eğilimi artmaktadır. Nitrik oksit (NO), özellikle trombositten zengin trom¬buslara yeni trombositlerin katılmasını sınırlandırarak trombosit fonksi¬yonlarında önemli rol oynar. Ancak, NO hücrelerde oksidatif/nitrozatif stresi artırarak zararlı etki de gösterebilir. Bu çalışmada, istirahat halinde ve kollajenle uyarılmış trombositlerde akut yüksek glikozun trombosit agregasyonuna, oksidatif stres parametrelerine ve NO biyoyararlanımına etkileri araştırılmıştır. Ayrıca, kuersetin (süperoksit anyon süpürücüsü) kullanılarak yüksek glikozun NO biyoyararlanımı üzerindeki etkilerinde süperoksit üretiminin rolü incelenmiştir. Yöntemler: Yıkanmış trombositler 5 mM D-glukoz (fizyolojik konsant¬rasyon, n:7) veya 25 mM D-glukoz (patofizyolojik konsantrasyon, n:7) veya kuersetin (10 μM) + 25 mM D-glukoz (n:7) ile 1 saat inkübe edildi. İnkübasyon sonrası, trombositlerde süperoksit üretimi, lipid peroksidas¬yonu (LPO), NO, nitrotirozin (NT) düzeyleri ve ilerlemiş glikasyon son ürünleri (AGE) ölçüldü. Ayrıca trombosit agregasyonu da incelendi. Bulgular: Dinlenme halinde ve kollajenle aktive trombositlerin yüksek glukoz ile inkübasyonu LPO, NT ve AGE düzeylerinde 5mM glukoz konsantrasyonuna göre istatistiksel olarak anlamlı artışlara neden oldu (p<0.05). Yüksek glukoz kollajenle aktive trombositlerde trombosit agregasyonunu, süperoksit oluşumunu ve NO düzeylerini anlamlı olarak artırdı (p<0.05). Yüksek glukoz ile inkübe edilen trombositlerde, kuer¬setin anlamlı olarak NO üretiminin ve NO biyoyararlanımının artmasına neden olurken oksidatif stresi baskıladı (p<0.05). Sonuç: Yüksek glukoz istirahat halinde ve özellikle kollajen ile aktive trombositlerde oksidatif stresi arttırır ve NO biyoyararlanımını azaltır. Yüksek glukoz aracılı süperoksit üretiminin bu etkiler üzerinde rolü olabi¬lir. Bulgularımız, diyabette trombosit-ilişkili komplikasyonların önlenme¬sinde kuersetinin yararlı etkisi olabileceğini göstermektedir.

Anahtar Kelimeler:

Hiperglisemi, kuersetin, kollajen, nitrik oksit, superoksit anyonu, trombosit

The effects of acute hyperglycemia on oxidative stress and nitric oxide bioavailability in platelets

Objective: Platelet activation and thrombus formation tendency increase in diabetes mellitus. Nitric oxide (NO) synthesized by platelets plays important role in platelet functions and limits the recruitment of new platelets to the platelet-rich thrombus. However, NO may also be deleterious by elevating oxidative/nitrosative stress in cells. The aim of the present study was to examine effects of acute high glucose in both resting and collagen-stimulated platelets on platelet aggregation, oxidative stress and NO bioavailability. In addition, we investigated the role of superoxide production on NO bioavailability using quercetin as scavengers of superoxide anion. Methods: Washed platelets were incubated with 5mM D-glucose (physiological concentration, n:7) or 25mM D-glucose (pathophysiological concentration, n:7) or quercetin (10 μM) plus 25 mM D-glucose (n:7) for 1 h. Superoxide production, lipid peroxidation (LPO), NO, nitrotyrosine (NT) levels and advanced glycation end products (AGE) in platelets were measured after incubation. Platelet aggregation was also investigated. Results: Incubation of resting and collagen-activated platelets with high glucose resulted in significant elevations in LPO, NT and AGE levels when compared to 5mM glucose concentration (p<0.05). High glucose significantly increased platelet aggregation, superoxide formation and NO levels in collagen-activated platelets (p<0.05). Quercetin markedly increased production and bioavailability of NO and suppressed oxidative stress in high glucose incubated platelets (p<0.05). Conclusion: High glucose increases oxidative stress and reduces bioavailability of NO in resting and especially collagen activated platelets. Hyperglycemia mediated superoxide production could be taken part on these effects. These findings would be suggested that there might be beneficial effects of quercetin in the prevention of platelet-related complications in diabetes mellitus.

Keywords:

Hyperglycemia, quercetin, collagen, nitric oxide, superoxide anion,

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