P2Y12 antagonist attenuates eosinophilic inflammation and airway hyperresponsiveness in a mouse model of asthma

Abstract

Leukotriene E4 (LTE4) that plays a key role in airway inflammation is expressed on platelets and eosinophils. We investigated whether blocking of the P2Y12 receptor can suppress eosinophilic inflammation in a mouse model of asthma because platelets and eosinophils share this receptor to be activated. BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA), followed by OVA nebulization. On each challenge day, clopidogrel, a P2Y12 antagonist was administered 30 min. before each challenge. Forty‐eight hours after the last OVA challenge, mice were assessed for airway hyperresponsiveness (AHR), cell composition and cytokine levels, including chemokine ligand 5 (CCL5), in bronchoalveolar lavage (BAL) fluid. EOL cells were treated with LTE4, with or without clopidogrel treatment, and intracellular and extracellular eosinophil cationic protein (ECP) expressions were measured to find the inhibiting function of P2Y12 antagonist on eosinophilic activation. The levels of P2Y12 expression were increased markedly in the lung homogenates of OVA‐sensitized and ‐challenged mice after platelet depletion. Administration of clopidogrel decreased AHR and the number of airway inflammatory cells, including eosinophils, in BAL fluid following OVA challenge. These results were associated with decreased levels of Th2 cytokines and CCL5. Histological examination showed that inflammatory cells as well as mucus‐containing goblet cells were reduced in clopidogrel‐administered mice compared to vehicle‐treated mice. Clopidogrel inhibited extracellular ECP secretion after LTE4 stimulation in EOL‐1 cells. Clopidogrel could prevent development of AHR and airway inflammation in a mouse model of asthma. P2Y12 can be a novel therapeutic target to the suppression of eosinophils in asthma.

The synergistic effects of clopidogrel with montelukast may be beneficial for asthma treatment

“…We used considerably lower doses of Mon and Clo (10 mg/kg for each drug) compared to previous experimental studies. 18,26,27 The secondary, eosinophilic asthma mouse model manifested higher eosinophilic infiltration than the model that we used previously to clearly detect the therapeutic effects of Clo/Mon. The Clo/Mon treatment suppressed the lymphocyte count in BAL fluid, which explained the decreased levels of Th2 cytokines.…”

“…16 Platelets and eosinophils share similar surface receptors, including the cysteinyl LT receptor type 1 (CysLTR1) and the P2Y12 purinergic receptor (P2Y12R). 18 Furthermore, LTE 4 , a P2Y12R and CysLTR1 agonist, induces eosinophilic inflammation following a platelet-dependent mechanism. 17 Our recent study suggested an inhibitory effect of clopidogrel (Clo), an antiplatelet drug that targets the P2Y12R, during eosinophilic inflammation and degranulation.…”

“…17 Our recent study suggested an inhibitory effect of clopidogrel (Clo), an antiplatelet drug that targets the P2Y12R, during eosinophilic inflammation and degranulation. 18 Furthermore, LTE 4 , a P2Y12R and CysLTR1 agonist, induces eosinophilic inflammation following a platelet-dependent mechanism. 19 We have suggested that LTE 4 amplifies the activation of platelets and eosinophils, thereby initiating the PEA and subsequent airway inflammation.…”

P2Y12 Inhibition beyond Thrombosis: Effects on Inflammation

“…In a clinical study involving cardiologic patients, Diehl et al showed that P2Y 12 mRNA was expressed in leukocytes obtained from leukapheresis and that P2Y 12 inhibition by clopidogrel decreased leukocyte CD11b (Mac-1) expression [59]. The P2Y 12 expression was also reported in human eosinophils in which P2Y 12 activation induced the release of eosinophil peroxidase [21] that could be prevented by clopidogrel [60]. Activation of P2Y 12 in macrophages induced cell spreading with formation of lamellipodia, and P2Y 12 inhibition alleviated chemotaxis [25].…”

P2Y12 Receptor Antagonist Clopidogrel Attenuates Lung Inflammation Triggered by Silica Particles

“…Furthermore, blocking P2Y 12 pathways alters T cell activation and changes the cell population (Vemulapalli et al, 2019). In the respiratory system, P2Y 12 receptor appears to contribute to inflammatory response, participating in allergic and non-allergic processes (Paruchuri et al, 2009;Shirasaki et al, 2013;Suh et al, 2016), as well as autoimmune disease processes (Domercq et al, 2018). Therefore, to better understand the role of purinergic signaling in silica-induced lung inflammation, we investigated the participation of P2Y 12 receptor in the onset of silicosis.…”