PAD4特異的阻害剤GSK484は新生児低酸素虚血性脳損傷において神経保護をもたらす

抄録

OBJECTIVE: To investigate whether the PAD4 specific inhibitor GSK484 ameliorates brain injury and neurological deficits by inhibiting NETs formation after HIBI and explore the underlying mechanisms.

METHODS: The classical Rice-Vannucci method was used to generate HIBI model. Mice were intraperitoneally injected with GSK484, and the administration time points were 48, 24, 0 h before HI induction and 24, 48 h after HI induction. The mice were divided into three groups: Sham, HI + Saline, and HI + GSK484. MWM, OF and EPM test were used to assess changes in cognitive and psychiatric function in mice. HE and Nissl staining were used to observe the pathological changes in brain tissue and neuronal structural damage. qRT-PCR was used to detect inflammatory factors and chemokines changes. WB and IF were used to demonstrate the expression levels of proteins associated with NETs.

RESULTS: GSK484 reversed HI insult-induced neurobehavioural deficits and the severity of brain injury; The results of QP revealed that the expression of inflammatory factors peaked in brain after 24 h of HI insult, but decreased substantially after the administration of GSK484; HE and Nissl staining showed that GSK484 ameliorated the morphological and structural deformation of brain and neuronal damage induced by HI insult. The mRNA level of chemokines increased significantly after HI insult, and GSK484 treatment reversed this change, suggesting GSK484 reduced neutrophil infiltration into brain; WB and IF indicated that GSK484 could significantly reduce the expression level of NETs-related proteins after HI insults, predicting that the mechanism of neuroprotective action of GSK484 may be through inhibition of NETs formation.

CONCLUSIONS: NETs promote neuroinflammation and brain dysfunction after HI insult, and GSK484-mediated pharmacological inhibition of NETs may provide neuroprotection. These data confirm that the PAD4-specific inhibitor GSK484 may be a potential therapeutic candidate for HIE.