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We previously demonstrated that ethanol-induced disruption of barrier function in human brain endothelial cells was associated with mitochondrial injury, increased ATP and extracellular vesicle (EV) release, and purinergic receptor P2\u2009\u00d7\u20097R activation. Therefore, we aimed to evaluate the effect of P2\u2009\u00d7\u20097R blockade on peripheral and neuro-inflammation in ethanol-exposed mice. In a chronic intermittent ethanol (CIE)-exposed mouse model, P2\u2009\u00d7\u20097R was inhibited by two different methods: Brilliant Blue G (BBG) or gene knockout. We assessed blood ethanol concentration (BEC), brain microvessel gene expression by using RT2 PCR array, plasma P2\u2009\u00d7\u20097R and P-gp, serum ATP, EV-ATP, number of EVs, and EV mtDNA copy numbers. An RT2 PCR array of brain microvessels revealed significant upregulation of proinflammatory genes involved in apoptosis, vasodilation, and platelet activation in CIE-exposed wild-type animals, which were decreased 15\u201350-fold in BBG-treated\u2013CIE-exposed animals. Plasma P-gp levels and serum P2\u2009\u00d7\u20097R shedding were significantly increased in CIE-exposed animals. Pharmacological or genetic suppression of P2\u2009\u00d7\u20097R decreased receptor shedding to levels equivalent to those in control group. The increase in EV number and EV-ATP content in the CIE-exposed mice was significantly reduced by P2\u2009\u00d7\u20097R inhibition. CIE mice showed augmented EV-mtDNA copy numbers which were reduced in EVs after P2\u2009\u00d7\u20097R inhibition or receptor knockout. These observations suggested that P2\u2009\u00d7\u20097R signaling plays a critical role in ethanol-induced brain injury. Increased extracellular ATP, EV-ATP, EV numbers, and EV-mtDNA copy numbers highlight a new mechanism of brain injury during alcohol exposure via P2\u2009\u00d7\u20097R and biomarkers of such damage. In this study, for the first time, we report the in vivo involvement of P2\u2009\u00d7\u20097R signaling in CIE-induced brain injury.<\/jats:p>","DOI":"10.1186\/s12974-024-03230-4","type":"journal-article","created":{"date-parts":[[2024,9,28]],"date-time":"2024-09-28T08:01:50Z","timestamp":1727510510000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["Neuroinflammatory responses and blood\u2013brain barrier injury in chronic alcohol exposure: role of purinergic P2\u2009\u00d7\u20097 Receptor signaling"],"prefix":"10.1186","volume":"21","author":[{"given":"Namdev S.","family":"Togre","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naveen","family":"Mekala","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Priyanka S.","family":"Bhoj","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikhita","family":"Mogadala","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malika","family":"Winfield","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jayshil","family":"Trivedi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah","family":"Grove","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sudhir","family":"Kotnala","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Slava","family":"Rom","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uma","family":"Sriram","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuri","family":"Persidsky","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,9,28]]},"reference":[{"key":"3230_CR1","doi-asserted-by":"crossref","unstructured":"Naimi MBEASYLTS. 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