Inflammation is extremely important in the development of
cerebral ischemia/
reperfusion injury. Pannexin 1 (Panx1) channel has been reported to activate
inflammasome in astrocytes and be involved in ischemic injury, but this damage effect is reversed by a Panx1 inhibitor-
probenecid. However, the mechanism of
probenecid protects against
cerebral ischemia/
reperfusion injury remains unclear. In present study, we hypothesized that
probenecid protected astrocytes from
ischemia/reperfusion injury in vitro by modulating the
inflammasome. Primary cultured neocortical astrocytes were exposed to
oxygen-
glucose deprivation/reoxygenation (OGD/RX) and
probenecid was added in this model. Viability and nuclear morphology of astrocytes, production of
reactive oxygen species (ROS),
protein expressions of NLRP3 (
NOD-like receptor protein 3), caspase-1, and AQP4 (
Aquaporins 4), as well as release of cellular
HMGB1 and IL-1β were observed to evaluate the effect and mechanisms of
probenecid on OGD/reoxygenated astrocytes.
Probenecid did not affect cell viability at concentrations of 1, 5, 10, and 100μM but induced significant astrocytes death at 500μM.
Probenecid inhibited cell death and ROS generation in astrocytes subjected to 6h of OGD and 24h of reoxygenation. The expression levels of NLRP3, caspase-1, and AQP4 increased after 6h of OGD, but
probenecid treatment attenuated this increase. Moreover, the extracellular release of IL-1β and
HMGB1 from OGD/reoxygenated astrocytes increased significantly. However, treatment by
probenecid resulted in substantial reduction of these
proteins levels in extracellular space. In conclusion, The Panx1 inhibitor,
probenecid, which was administered before OGD, provided protective effects on the OGD/reoxygenation model of cultured astrocytes by modulating
inflammasome activity and downregulating AQP4 expression.