Newborns suffering from hypoxia-ischemia(HI)brain injury still lack effective treatment.Proline-rich tyrosine kinase 2(Pyk2)is a non-receptor tyrosine kinase,which is highly correlated with transient ischemic brain injury in adult.In this study,we investigated the role of Pyk2 in neonatal HI brain injury.HI was induced in postnatal day 7 mouse pups by unilateral common carotid artery ligation followed by hypoxic exposure.Pyk2 interference lentivirus(LV-Pyk2 shRNA)was constructed and injected into unilateral cerebral ventricle of neonatal mice before HI.Infarct volume,pathological changes,and neurological behaviors were assessed on postnatal day 8-14.We showed that the phosphorylation level of Pyk2 was significantly increased in neonatal brain after HI,whereas LV-Pyk2 shRNA injection significantly attenuated acute HI brain damage and improved neurobehavioral outcomes.In oxygen-glucose deprivation-treated cultured cortical neurons,Pyk2 inhibition significantly alleviated NMDA receptor-mediated excitotoxicity;similar results were also observed in neonatal HI brain injury.We demonstrated that Pyk2 inhibition contributes to the long-term cerebrovascular recovery assessed by laser speckle contrast imaging,but cognitive function was not obviously improved as evaluated in Morris water maze and novel object recognition tests.Thus,we constructed lentiviral LV-HIF-Pyk2 shRNA,through which HIF-1 α promoter-mediated interference of Pyk2 would occur during the anoxic environment.Intracerebroventricular injection of LV-HIF-Pyk2 shRNA significantly improved long-term recovery of cognitive function in HI-treated neonatal mice.In conclusion,this study demonstrates that Pyk2 interference protects neonatal brain from hypoxic-ischemic injury.HIF-1 α promoter-mediated hypoxia conditional control is a useful tool to distinguish between hypoxic period and normal period.Pyk2 is a promising drug target for potential treatment of neonatal HI brain injury.

Jie Zhu;Shi-feng Chu;Ye Peng;Dan-dan Liu;Chen Chen;Wen-xuan Jian;Hong-shuo Sun;Zhong-ping Feng;Zhao Zhang;Nai-hong Chen

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines,Institute of Materia Medica&Neuroscience Center,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China;College of Pharmacy,Hunan University of Chinese Medicine,Changsha 410208,China;Institute of Clinical Pharmacology,Guangzhou University of Chinese Medicine,Guangzhou 510720,China;Department of Physiology,Faculty of Medicine,University of Toronto,Toronto,ON,Canada

中国药理学报（英文版）

[1]Jie Zhu;Shi-feng Chu;Ye Peng;Dan-dan Liu;Chen Chen;Wen-xuan Jian;Hong-shuo Sun;Zhong-ping Feng;Zhao Zhang;Nai-hong Chen-.Pyk2 inhibition attenuates hypoxic-ischemic brain injury in neonatal mice)[J].中国药理学报（英文版）,2022(04):797-810

Newborns,pups,Intracerebroventricular

Pyk2,inhibition,attenuates,hypoxic,ischemic,brain,injury,neonatal,mice,suffering,from,hypoxia,ischemia,still,lack,effective,treatment,Proline,rich,tyrosine,kinase,receptor,which,highly,correlated,transient,adult,this,study,investigated,role,was,induced,postnatal,day,mouse,by,unilateral,common,carotid,artery,ligation,followed,exposure,interference,lentivirus,LV,shRNA,constructed,injected,into,cerebral,ventricle,before,Infarct,volume,pathological,changes,neurological,behaviors,were,assessed,We,showed,that,phosphorylation,level,significantly,increased,after,whereas,injection,attenuated,acute,damage,improved,neurobehavioral,outcomes,oxygen,glucose,deprivation,treated,cultured,cortical,neurons,alleviated,NMDA,mediated,excitotoxicity,similar,results,also,observed,demonstrated,contributes,long,term,cerebrovascular,recovery,laser,speckle,contrast,imaging,cognitive,function,not,obviously,evaluated,Morris,water,maze,novel,object,recognition,tests,Thus,lentiviral,HIF,through,promoter,would,occur,during,anoxic,environment,conclusion,demonstrates,protects,conditional,control,useful,tool,distinguish,between,period,normal,promising,drug,target,potential

0.513247