Neuronal chemokine-like-factor 1(CKLF1)up-regulation promotes M1 polarization of microglia in rat brain after stroke  

作　　者：Xin Zhou Ya-ni Zhang Fang-fang Li Zhao Zhang Li-yuan Cui Hong-yuan He Xu Yan Wen-bin He Hong-shuo Sun Zhong-ping Feng Shi-feng Chu Nai-hong Chen 

机构地区：[1]State Key Laboratory of Bioactive Substances and Functions of Natural Medicines,Institute of Materia Medica and Neuroscience Center,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing,100050,China [2]Institute of Clinical Pharmacology&Science and Technology Innovation Center,Guangzhou University of Chinese Medicine,Guangzhou,510405,China [3]Tianjin University of Tradition Chinese Medicine,Tianjin,301617,China [4]Shanxi Key Laboratory of Chinese Medicine Encephalopathy,Shanxi University of Chinese Medicine,Jinzhong 030619,China [5]Department of Physiology,Faculty of Medicine,University of Toronto,Toronto,ON,Canada

出　　处：《Acta Pharmacologica Sinica》2022年第5期1217-1230,共14页中国药理学报（英文版）

基　　金：This work was supported by the National Natural Science Foundation of China(81730096,82074044,81873026,81773924);the Natural Science Foundation of Beijing(7192135);the Drug Innovation Major Project(2018ZX09711001-002-007,2018ZX09711001-003-005,2018ZX09711001-009-013);the CAMS Innovation Fund for Medical Sciences(CIFMS)(2016-I2M-1-004);the High-End Foreign Experts Introduction Program(G20200001485).

摘　　要：The phenotypic transformation of microglia in the ischemic penumbra determines the outcomes of ischemic stroke.Our previous study has shown that chemokine-like-factor 1(CKLF1)promotes M1-type polarization of microglia.In this study,we investigated the cellular source and transcriptional regulation of CKLF1,as well as the biological function of CKLF1 in ischemic penumbra of rat brain.We showed that CKLF1 was significantly up-regulated in cultured rat cortical neurons subjected to oxygen-glucose deprivation/reoxygenation(ODG/R)injury,but not in cultured rat microglia,astrocytes and oligodendrocytes.In a rat model of middle cerebral artery occlusion,we found that CKLF1 was up-regulated and co-localized with neurons in ischemic penumbra.Furthermore,the up-regulated CKLF1 was accompanied by the enhanced nuclear accumulation of NF-κB.The transcriptional activity of CKLF1 was improved by overexpression of NF-κB in HEK293T cells,whereas application of NF-κB inhibitor Bay 11-7082(1μM)abolished it,caused by OGD/R.By using chromatin-immunoprecipitation(ChIP)assay we demonstrated that NF-κB directly bound to the promoter of CKLF1(at a binding site located at−249 bp to−239 bp of CKLF1 promoter region),and regulated the transcription of human CKLF1.Moreover,neuronal conditional medium collected after OGD/R injury or CKLF1-C27(a peptide obtained from secreted CKLF1)induced the M1-type polarization of microglia,whereas the CKLF1-neutralizing antibody(αCKLF1)or NF-κB inhibitor Bay 11-7082 abolished the M1-type polarization of microglia.Specific knockout of neuronal CKLF1 in ischemic penumbra attenuated neuronal impairments and M1-type polarization of microglia caused by ischemic/reperfusion injury,evidenced by inhibited levels of M1 marker CD16/32 and increased expression of M2 marker CD206.Application of CKLF1-C27(200 nM)promoted the phosphorylation of p38 and JNK in microglia,whereas specific depletion of neuronal CKLF1 in ischemic penumbra abolished ischemic/reperfusion-induced p38 and JNK phosphorylation.In summary

关 键 词：STROKE ischemic penumbra CKLF1 microglia polarization cortical neurons NF-κB inflammation 

分 类 号：R96[医药卫生—药理学]