The role of the P2X7 receptor in inactivated SARS-CoV-2-induced lung injury

Purinergic signaling plays a role in the pathophysiology of different viral infections. Recently, we showed that COVID-19 increases extracellular ATP levels, which may amplify the pro-inflammatory signals in the disease. The P2X7 receptor can be a protagonist in the pro-inflammatory responses. Herein, we investigated the role of the P2X7 receptor in the lung immune response triggered by inoculation of inactivated SARS-CoV-2 (iSARS-CoV-2) in K18-Human ACE2 transgenic mice. Pharmacological inhibition of the P2X7 receptor was performed with intraperitoneal administration of 50 mg/kg of Brilliant Blue G (BBG) one day before viral inoculation. Animals were divided into four groups: a control group (MOCK), a group inoculated with the inactivated virus iSARS-CoV-2, a BBG-treated control group (MOCK + BBG), and a BBG-treated inoculated group (iSARS-CoV-2 + BBG). Virus inoculation was intratracheal with 50 µl of mock or 2 × 10⁶ Plaque Forming Units (PFU) of iSARS-CoV-2. After three days, blood and lungs were collected. We found a significant increase in ATP and LDH in serum and mRNA levels of P2X7 and P2Y₁₂ receptors, CD39, IL-1β, and TNF-α in the lung of the iSARS-CoV-2 group when compared with the control group. BBG treatment attenuated these increases. Lung histological analyses showed severe lung damage in the iSARS-CoV-2 group, which was reduced by the BBG treatment. Immunohistochemical staining confirmed the increased presence of P2X7, P2Y₁₂, and CD39 proteins in the iSARS-CoV-2 vs. the MOCK group. Thus, P2X7 receptor inhibition decreases iSARS-CoV-2-induced lung inflammation, indicating that this receptor might contribute to SARS-CoV-2 pathology.

iSARS-CoV-2 modulates purinergic signaling and exacerbates the pro-inflammatory response, resulting in lung damage. The resulting release of ATP into the extracellular environment triggers events that enhance the activation of P2X7 and P2Y₁₂ receptors and CD39. This activation may be associated with increased production of pro-inflammatory cytokines, including IL-1β and TNF-α, which causes extensive lung damage. These data suggest that inhibition of P2X7 receptor signaling ameliorates lung inflammation, preventing lung injury. iSARS (inactivated SARS-CoV-2); BBG (Brilliant Blue G). (Created with BioRender.com).