机构：[1]Department of Cerebrovascular Diseases, The Second Affiliated Hospital of Kunming Medical University, Kunming, China[2]Department of Neurology, The Second Affiliated Hospital of Kunming Medical University, Kunming, China[3]Department of Radiology, The Second Affiliated Hospital of Kunming Medical University, Kunming, China[4]Department of Neurosurgery, The Second Affiliated Hospital of Kunming Medical University, Kunming, China[5]Department of Anesthesiology, Huashan Hospital, Fudan University, Shanghai, China[6]Department of Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing, China神经科系统神经外科首都医科大学宣武医院[7]International Neuroscience Institute (China-INI), Xuanwu Hospital, Capital Medical University, Beijing, China首都医科大学宣武医院[8]Department of Cardiology, The Second Affiliated Hospital of Kunming Medical University, Kunming, China
Human brain microvascular endothelial cells (HBMVECs) and microglia play critical roles in regulating cerebral homeostasis during ischemic stroke. However, the role of HBMVECs-derived exosomes in microglia polarization after stroke remains unknown. We isolated exosomes (Exos) from oxygen glucose deprivation (OGD)-exposed HBMVECs, before added them into microglia. Microglia polarization markers were tested using RT-qPCR or flow cytometry. Inflammatory cytokines were measured with ELISA. Endothelial cell damage was assessed by cell viability, apoptosis, apoptosis-related proteins, oxidative stress, and angiogenic activity using CCK-8, flow cytometry, western blot, ELISA, and endothelial tube formation assay, respectively. We also established middle cerebral artery occlusion (MCAO) mice model to examine the function of circ_0000495 on stroke in vivo. Our study found that HBMVECs-Exos reduced M2 markers (IL-10, CD163, and CD206), increased M1 markers (TNF-alpha, IL-1 beta, and IL-12), CD86-positive cells, and inflammatory cytokines (TNF-alpha and IL-1 beta), indicating the promotion of microglial M1-polarization. Microglial M1-polarization induced by HBMVECs-Exos reduced viability and promoted apoptosis and oxidative stress, revealing the aggravation of endothelial cell damage. However, circ_0000495 silencing inhibited HBMVECs-Exos-induced alterations. Mechanistically, circ_0000495 adsorbed miR-579-3p to upregulate toll-like receptor 4 (TLR4) in microglia; miR-579-3p suppressed HBMVECs-Exos-induced alterations via declining TLR4; furthermore, Yin Yang 1 (YY1) transcriptionally activated circ_0000495 in HBMVECs. Importantly, circ_0000495 aggravated ischemic brain injury in vivo via activating TLR4/nuclear factor-kappa B (NF-kappa B) pathway. Collectively, OGD-treated HBMVECs-Exos transmitted circ_0000495 to regulate miR-579-3p/TLR4/NF-kappa B axis in microglia, thereby facilitating microglial M1-polarization and endothelial cell damage.
基金：
The National Natural ScienceFundation of China, Grant/AwardNumber: 82260246, 81960219; The Social Development SpecializedProject of Yunnan ProvincialKey Technology Research andDevelopment Program, Grant/AwardNumber: 202203AC100007; YunnanProvincial Applied Basic Researchjoint project, Grant/Award Number:202301AY070001-271; Academic leaderof the talent team project of the SecondAffiliated Hospital of Kunming MedicalUniversity, Grant/Award Number:RCTDXS-202307
第一作者机构：[1]Department of Cerebrovascular Diseases, The Second Affiliated Hospital of Kunming Medical University, Kunming, China
通讯作者：
通讯机构：[5]Department of Anesthesiology, Huashan Hospital, Fudan University, Shanghai, China[6]Department of Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing, China[7]International Neuroscience Institute (China-INI), Xuanwu Hospital, Capital Medical University, Beijing, China[8]Department of Cardiology, The Second Affiliated Hospital of Kunming Medical University, Kunming, China[*1]Department of Cardiology, The Second AffiliatedHospital of Kunming MedicalUniversity, Kunming 650101, Yunnan,China[*2]Department of Neurosurgery & International Neuroscience Institute(China-INI), Xuanwu Hospital, CapitalMedical University, Beijing 100053,China[*3]Department of Anesthesiology,Huashan Hospital, Fudan University,Shanghai 200040, China
推荐引用方式(GB/T 7714)：
Min Xiao-li,Jia Wen-ji,Guo Li,et al.Brain microvascular endothelial cell-derived exosomes transmitting circ_0000495 promote microglial M1-polarization and endothelial cell injury under hypoxia condition[J].FASEB JOURNAL.2024,38(2):doi:10.1096/fj.202301637R.
生物学