Neural cells and tissue injury lead to abundant release of ATP from the cell. Additionally, ATP could be cut loose through the activated Pannexin 1 hemichannels. Extracellular ATP could activate P2X7R, P2 ligand-gated ion channels, activating the inflammasome in microglia and astrocytes. P2X7R and Panx1 could create forward feedback loop, that potentiates and drives the chronicity of previously initiated neuroinflammation. Both P2X7R and Panx1 are upregulated in multiple neurodegenerative diseases, hypoxia, injuries, depression, epilepsy, and pain. Targeting one or both P2X7R and Panx1 has become attractive therapeutic strategy in neuroinflammation, in COVID-19-associated inflammation, and in cancer as well. Nevertheless, there is a need to understand and define the effects of P2X7R and/or Panx1 inhibition, especially in astrocytes, which are the major contributors and regulators of neuroinflammation.In this study the inhibitors of P2X7R (A438079), and Panx1 (Probenecid) were applied in an in vitro astrocyte activation/injury model: scratch wound with 1 mM ATP addition. Our results show that by blocking the P2X7R, markers of activated astrocytes (C3, IL-1β, LCN2) are downregulated to control levels. The similar effect was achieved by applying Probenecid, which disturbed the Panx1-ATP-P2X7-inflamasome inflammatory loop. The ATP downstream signaling purinergic components were analyzed as well: ecto-nucleotidases (CD73) and adenosine receptors (A1 and A2), since they play important role in neuroinflammation regulation. Both A438079 and Probenecid induced significant astrocyte antioxidative protection. Moreover, our results suggest that Probenecid, an FDA-approved drug, should be reconsidered as an anti-neuroinflammatory agent. Acknowledgement:Ministry of Science, Technological Development and Innovation RS 451-03-66/2024-03/ 200178