机构：[1]Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, P.R. China[2]Department of Neurology, The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming 650034, Yunnan, P.R. China内科片神经内科云南省第一人民医院[3]Department of Magnetic Resonance, The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming 650034, Yunnan, P.R. China医技片磁共振科云南省第一人民医院[4]Department of Geriatric Medicine, The First People’ s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming 650034, Yunnan, P.R. China云南省第一人民医院
Background: Hemorrhagic transformation (HT) caused by blood-brain barrier (BBB) damage is closely correlated with the poor prognosis of ischemic stroke. Neutrophils are proven to mediate BBB injury after ischemic stroke, but the mechanism remains to be further investigated. Therefore, the present study aims to investigate the effect of neutrophil-derived exosomes on BBB integrity. Method: A tMCAO-HT model was constructed to assess neutrophil infiltration and its co-localization with brain microvascular endothelial cells (BMEC). After using quiet (Q-Neu) and activated neutrophil (A-Neu) and their exosomes to treat the BBB model in vitro, TEER and permeability were assayed to assess the BBB integrity. Small RNA sequencing was performed to identify differentially expressed miRNAs (DE-miRNAs) in A-Neu- and Q-Neuderived exosomes, and the function and pathways of DE-miRNA targets were analyzed by GO and KEGG enrichment. Result: Different degrees of cerebral hemorrhage were observed in the tMCAO-HT model. The expression of the neutrophil marker Ly6G was significantly increased in tMCAO-HT model compared to the sham group, and colocalized with the BMEC marker CD31. Notably, Ly6G expression was positively correlated with hemoglobin content in brain tissue. A-Neu and its derived exosomes reduced TEER and elevated permeability in the BBB model in vitro. Moreover, BBB-related proteins Claudin 5, Occludin and ZO-1 expression were significantly reduced in BMEC after treatment with A-Neu and its derived exosomes. Nevertheless, Q-Neu and its exosomes had no significant effect on BBB integrity. A total of 84 DE-miRNAs are present in Q-Neu- and A-Neu-derived exosomes, and their target genes are involved in the regulation of "positive regulation of establishment of endothelial barrier", "cell junction", "ECM-receptor interaction" and "VEGF signaling pathway". Moreover, RTqPCR revealed that the expression trends of miR-409-3p, miR-6909-5p, miR-3473d, miR-370-3p and miR-69045p in exosomes were consistent with the sequencing results. Conclusion: Neutrophils are abnormally recruited in HT after ischemic stroke, and are associated with cerebral hemorrhage. In vitro, A-Neu-derived exosomes facilitate BBB injury, which may be accomplished by exosomal transport of miRNAs.
基金：
National Natural Science Foundation of China [81960228]; Yunnan health training project of high level talents [L-2017013]; ten thousand talents program-famous doctor project of Yunnan Province [YNWR-MY-2018-018]
第一作者机构：[1]Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, P.R. China[2]Department of Neurology, The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming 650034, Yunnan, P.R. China
通讯作者：
通讯机构：[1]Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, P.R. China[*1]Department of Neurology, The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, No. 157 Jinbi Road, Xishan District, Kunming 650032, Yunnan Province, China.
推荐引用方式(GB/T 7714)：
Tang Ning,Gong Xia-Rong,Huang Hong,et al.Activated neutrophil-derived exosomes contribute to blood-brain barrier damage and hemorrhagic transformation after cerebral ischemia/ reperfusion[J].BRAIN RESEARCH.2023,1810:doi:10.1016/j.brainres.2023.148374.
APA：
Tang, Ning,Gong, Xia-Rong,Huang, Hong&Meng, Qiang.(2023).Activated neutrophil-derived exosomes contribute to blood-brain barrier damage and hemorrhagic transformation after cerebral ischemia/ reperfusion.BRAIN RESEARCH,1810,
MLA：
Tang, Ning,et al."Activated neutrophil-derived exosomes contribute to blood-brain barrier damage and hemorrhagic transformation after cerebral ischemia/ reperfusion".BRAIN RESEARCH 1810.(2023)
