Abstract
The pathogenesis of neonatal hypoxic-ischemic (HI) brain injury may involve activation of the NOD-like receptor family pyrin domain-containing-3 (NLRP3) inflammasome and its downstream effectors, caspase-1 and interleukin (IL)-1β. The start time of therapy is associated with adverse neurodevelopmental outcome following HI injury. We performed this study investigating early dynamic changes in NLRP3, caspase-1, and IL-1β expression during the first 24 h following HI brain injury in an animal model, in order to optimize selection of treatment time after injury. Rats were randomized to an HI group (n=40) and sham group (n=40). Rats in the HI group were subjected to right common carotid artery ligation and then exposed to hypoxia (8% O2) for 2 h, and divided into 5 subgroups with 8 cases in each group at 5 postoperative time points (0, 4, 8, 12, 24 h). Brain injury during the first 24 h after surgery/hypoxia was evaluated by cranial ultrasonography. RT-PCR, western blot, and immunohistochemistry were applied to determine protein and mRNA expressions. In the HI group, ultrasonography revealed accelerated right vertebrobasilar artery flow at 4 h, enhanced brain parenchyma echogenicity at 24 h, and blood stealing from the vertebrobasilar artery at 24 h. In the HI group, immunohistochemistry demonstrated elevated expressions of NLRP3 and IL-1β at 4, 8, 12, and 24 h and enhanced expression of caspase-1 at 8 and 12 h (all P < 0.01). Western blot and RT-PCR revealed that, compared with the sham group, the HI group exhibited elevated expression of NLRP3 at 4, 8, and 24 h, caspase-1 at 12 h, and IL-1β at 8 h (all P < 0.05). In summary, the present results suggested that activation of NLRP3/caspase-1/IL-1β signaling occurs within 4 h of HI brain injury in the neonatal rat.