Pinar Ulker1, Nur Özen1, Günel Abdullayeva1, Sadi Köksoy2, Nazmi Yaraş3, Filiz Basrali1. 1. Department of Physiology, Medical Faculty, Akdeniz University, Antalya, Turkey. 2. Department of Medical Microbiology, Medical Faculty, AkdenizUniversity, Antalya, Turkey. 3. Department of Biophysics, Medical Faculty, AkdenizUniversity, Antalya, Turkey.
Abstract
BACKGROUND: It has been well documented that ATP activates NOS enzymes and causes increased NO production in several cell types. Although RBC known to possesses eNOS enzyme activity, it has not been investigated whether RBC eNOS could be induced by extracellular ATP. OBJECTIVE: The aim of the present study is to evaluate extracellular ATP mediated eNOS activation and NO production in RBC. METHODS: RBC packed were isolated from healthy volunteers and re-suspended in Hepes solution at a hematocrit of 0.01 l/l. Intracellular NO and Ca+2 levels and eNOS activation measured by flow cytometry in response to P2X receptor agonist, Bz-ATP, in the absence and presence of NOS, P2 receptors and PI3K inhibitors. RESULTS: P2X receptor agonist Bz-ATP found to increase intracellular NO, Ca+2 and serine 1177 phosphorylated eNOS levels and these responses have shown to be suppressed by NOS enzyme, P2 receptors and PI3K inhibitors. CONCLUSIONS: The results of the study clearly demonstrated extracellular ATP induced NO generation in RBC through intracellular Ca+2 and PI3K/Akt pathways. The mechanism we described here might be important at basal conditions and also in conditions with increased ATP release.
BACKGROUND: It has been well documented that ATP activates NOS enzymes and causes increased NO production in several cell types. Although RBC known to possesses eNOS enzyme activity, it has not been investigated whether RBC eNOS could be induced by extracellular ATP. OBJECTIVE: The aim of the present study is to evaluate extracellular ATP mediated eNOS activation and NO production in RBC. METHODS: RBC packed were isolated from healthy volunteers and re-suspended in Hepes solution at a hematocrit of 0.01 l/l. Intracellular NO and Ca+2 levels and eNOS activation measured by flow cytometry in response to P2X receptor agonist, Bz-ATP, in the absence and presence of NOS, P2 receptors and PI3K inhibitors. RESULTS: P2X receptor agonist Bz-ATP found to increase intracellular NO, Ca+2 and serine 1177 phosphorylated eNOS levels and these responses have shown to be suppressed by NOS enzyme, P2 receptors and PI3K inhibitors. CONCLUSIONS: The results of the study clearly demonstrated extracellular ATP induced NO generation in RBC through intracellular Ca+2 and PI3K/Akt pathways. The mechanism we described here might be important at basal conditions and also in conditions with increased ATP release.
Entities:
Keywords:
ATP; Red blood cells; nitric oxide; purinergic receptors
Authors: Tamas Alexy; Jon Detterich; Philippe Connes; Kalman Toth; Elie Nader; Peter Kenyeres; Jose Arriola-Montenegro; Pinar Ulker; Michael J Simmonds Journal: Front Physiol Date: 2022-07-06 Impact factor: 4.755