Esculetin facilitates post-stroke rehabilitation by inhibiting CKLF1-mediated neutrophil infiltration  

作　　者：Jia-qi He Ruo-lan Yuan Yu-tong Jiang Ye Peng Jun-rui Ye Sha-sha Wang Li-qing Li Yuan Ruan Pei-yi Li Xu Yan Wen-bin He Gang Li Shi-feng Chu Zhao Zhang Nai-hong Chen 

机构地区：[1]Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China [2]State Key Laboratory of Bioactive Substances and Functions of Natural Medicines,Institute of Materia Medical&Neuroscience Center,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China [3]National International Joint Research Center for Molecular Chinese Medicine,Shanxi University of Chinese Medicine,Taiyuan 030024,China [4]Graduate school,Inner Mongolian Medical University,Hohhot 010110,China

出　　处：《Acta Pharmacologica Sinica》2025年第1期52-65,共14页中国药理学报(英文版)

基　　金：This work was supported by the National Key R&D Program of China(2022YFC3500300);National Natural Science Foundation of China(U2202214,82074044,82374060,82130109,U21A20410);the CAMSInnovationFund for Medical Sciences(CIFMS)(2021-I2M-1-020).

摘　　要：Esculetin(ESC)is a coumarin-derived phytochemical prevalent in traditional Chinese medicine that exhibits anti-acute ischemic stroke activities.Our previous studies demonstrate that CKLF1 is a potential anti-stroke target for coumarin-derived compound.In this study we investigated whether CKLF1 was involved in the neuroprotective effects of ESC against photothrombotic stroke in mice.The mice were treated with ESC(20,40 or 80 mg.kg^(-1).d^(-1),i.g.)for two weeks.The therapeutic effect of ESC was assessed using MRl,neurological function evaluation,and a range of behavioral tests on D1,3,7 and 14 of ESC administration.We showed that oral administration of EsC dose-dependently reduced the cerebral infarction volume within one week after stroke,improved behavioral performance,and alleviated neuropathological damage within two weeks.Functional MRI revealed that ESC significantly enhanced the abnormal low-frequency fluctuation(ALFF)value of the motor cortex and promoted functional connectivity between the supplementary motor area(SMA)and multiple brain regions.We demonstrated that ESC significantly reduced the protein levels of CKLF1 and CCR5,as well as the CKLF1/CCR5 protein complex in the peri-infarcted area.We showed that ESC(0.1-10μM)dosedependently blocked CKLF1-induced chemotactic movement of neutrophils in the Transwell assay,reducing the interaction of CKLF1/CCR5 on the surface of neutrophils,thereby reducing neutrophil infiltration,and decreasing the expression of ICAM-1,VCAM-1 and MMP-9 in the peri-infarct tissue.Knockout of CKLF1 reduced brain infarction volume and motor dysfunction after stroke but also negated the anti-stroke efficacy and neutrophil infiltration of ESC.These results suggest that the effcacy of EsC in promoting post-stroke neural repair depends on its inhibition on CKLF1-mediated neutrophil infiltration,which offering novel perspectives for elucidating the therapeutic properties of coumarins.

关 键 词：ischemic stroke ESCULETIN COUMARIN neutrophil infiltration CKLF1 CCR5 

分 类 号：R74[医药卫生—神经病学与精神病学]