Pyk2 inhibition attenuates hypoxic-ischemic brain injury in neonatal mice  

作　　者：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 

机构地区：[1]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 [2]College of Pharmacy,Hunan University of Chinese Medicine,Changsha,410208,China [3]Institute of Clinical Pharmacology,Guangzhou University of Chinese Medicine,Guangzhou,510720,China [4]Department of Physiology,Faculty of Medicine,University of Toronto,Toronto,ON,Canada

出　　处：《Acta Pharmacologica Sinica》2022年第4期797-810,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(81730096,81873026,81973499);the CAMS Innovation Fund for Medical Sciences(CIFMS)(2016-I2M-1-004);the Drug Innovation Major Project(2018ZX09711001-003-005,2018ZX09711001-002-007);the Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study(BZ0150);the opening Program of Shanxi Key Laboratory of Chinese Medicine Encephalopathy(CME-OP-2017001);the High-End Foreign Experts introduction program(G20200001485).

摘　　要：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-1α 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-1α 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.

关 键 词：neonatal mice hypoxic-ischemic brain injury PYK2 NMDA receptor EXCITOTOXICITY HIF-1Α 

分 类 号：R722.1[医药卫生—儿科]