NLRP3 promotes radiation‐induced brain injury by regulating microglial pyroptosis

Abstract Purpose Radiation‐induced brain injury, one of the side effects of cranial radiotherapy in tumour patients, usually results in durable and serious cognitive disorders. Microglia are important innate immune‐effector cells in the central nervous system. However, the interaction between microglia and neurons in radiation‐induced brain injury remains uncharacterised. Methods and Materials We established a microglia–neuron indirect co‐culture model to assess the interaction between them. Microglia exposed to radiation were examined for pyroptosis using lactate dehydrogenase (LDH) release, Annexin V/PI staining, SYTOX staining and western blot. The role of nucleotide‐binding oligomerisation domain‐like receptor family pyrin domain containing 3 (NLRP3) was investigated in microglia exposed to radiation and in mouse radiation brain injury model through siRNA or inhibitor. Mini‐mental state examination and cytokines in blood were performed in 23 patients who had experienced cranial irradiation. Results Microglia exerted neurotoxic features after radiation in the co‐culture model. NLRP3 was up‐regulated in microglia exposed to radiation, and then caspase‐1 was activated. Thus, the gasdermin D protein was cleaved, and it triggered pyroptosis in microglia, which released inflammatory cytokines. Meanwhile, treatment with siRNA NLRP3 in vitro and NLRP3 inhibitor in vivo attenuated the damaged neuron cell and cognitive impairment, respectively. What is more, we found that the patients after radiation with higher IL‐6 were observed to have a decreased MMSE score. Conclusions These findings indicate that radiation‐induced pyroptosis in microglia may promote radiation‐induced brain injury via the secretion of neurotoxic cytokines. NLRP3 was evaluated as an important mediator in radiation‐induced pyroptosis and a promising therapeutic target for radiation‐induced brain injury.