Abstract WP236: The Gut Microbiome Is Associated With Early Brain Injury After Experimental Subarachnoid Hemorrhage Via Regulation Of Neutrophils

Abstract

Background: It is known that early brain injury (EBI), which occurs early after the onset of subarachnoid hemorrhage (SAH), largely determines the prognosis of SAH; however, an effective treatment for EBI has not been developed. The outcome of various diseases including ischemic stroke is influenced by the composition of the gut microbiome (GM) and we have previously shown that the GM is also associated with the rupture of cerebral aneurysms. Here, we studied whether prior alteration of the GM can prevent EBI after SAH. Methods: We altered the GM of 7-week-old male rats by administering antibiotics-containing water for 2 weeks or performing fecal microbiome transplantation after antibiotics treatment. The composition of the GM was profiled using 16S rRNA. SAH was induced by injection of blood in the subarachnoid space of control rats and rats with altered GM. We evaluated EBI indicators such as the neurological score, brain water content, Evans blue extravasation, and neuronal apoptosis. We also studied inflammatory cells using immunohistochemistry, immunocytochemistry, quantitative PCR and flow cytometry. Results: There were significant differences in alpha and beta diversity between the control and antibiotics-treated rats. Compared with the control rats, the antibiotics-treated rats had significantly reduced neurological symptoms, brain edema, blood-brain barrier disruption and apoptosis. Among inflammatory cells, neutrophil infiltration into the brain was significantly reduced in the antibiotics-treated rats. Interestingly, the preventive effect on neutrophil infiltration was already observed immediately after SAH. Next, the immune status of each group before SAH onset was examined. There was no difference in circulating neutrophil counts, but flow cytometry showed a significant decrease in aged neutrophils which exhibit a highly reactive phenotype in the antibiotics-treated rats. Finally, the neuroprotective effect of antibiotics against EBI disappeared when the GM was recolonized. Conclusions: Our findings suggest that the GM affects the severity of EBI after SAH by regulating neutrophils. In the future, adjustment of the GM to prevent EBI may become a new strategy in patients who are being followed for cerebral aneurysms.