The expression of Nod-like receptor protein 3 (NLRP3) in necroptosis of primary cortical neurons

ObjectiveTo investigate the levels of Nod-like receptor protein 3 (NLRP3) in primary cortical neurons with necroptosis. MethodsPrimary cortical neurons of SD rats were cultured for 12 days, before pretreated with Q-VD-Oph for 30min followed by oxygen-glucose deprivation to induce necroptosis. ① The cells were divided into the following groups: a normal control group, an oxygen-glucose deprivation group (oxygen-glucose deprivation for 2 h followed by reperfusion for 12 h), and treatment groups (oxygen-glucose deprivation for 2 h followed by reperfusion for 3, 6, 12 and 24 h, and all of them were treated with Q-VD-Oph) Their levels of NLRP3 and lactate dehydrogenase (LDH) were examined. ② The cells were divided into the following groups: a control group, an oxygen-glucose deprivation group (oxygen-glucose deprivation for 2 h followed by reperfusion for 12 h), and treatment groups (oxygen-glucose deprivation for 0.5, 1, 1.5, 2, and 3 h followed by reperfusion for 12 h, and all of them were treated with Q-VD-Oph). Their levels of NLRP3 and LDH were examined. ③ The cells were subjected to oxygen-glucose deprivation for 2 h followed by reperfusion for 12 h (all of them were treated with Q-VD-Oph) before determination of their levels of NLRP3 by immunofluorescence. ResultsAfter oxygen-glucose deprivation for 2 h, the level of NLRP3 was increased as the time of reperfusion extended, which then reached the highest at 12 h and was remarkably decreased at 24 h (P<0.05). The level of LDH reached the highest at 12 h. After oxygen-glucose deprivation for different periods of time followed by reperfusion for 12 h, the level of NLRP3 reached the highest when oxygen-glucose deprivation for 2 h, which then was remarkably decreased at 3 h (P<0.05). The level of LDH reached the highest when oxygen-glucose deprivation for 2 h. The immunofluorescence intensity was the highest when the cells were oxygen-glucose deprived for 2 h followed by reperfusion for 12 h. ConclusionsNLRP3 is present in primary cortical neuron with necroptosis. The degree of cell injury is gradually increased ass the level of NLRP3 increases.