Pyk2 inhibition attenuates hypoxic-ischemic brain injury in neonatal mice
Pyk2 inhibition attenuates hypoxic-ischemic brain injury in neonatal mice
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Pyk2 抑制可减轻新生小鼠缺氧缺血性脑损伤
DOI：
10.1038/s41401-021-00694-5
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发表时间：
2022
影响因子：
8.2
通讯作者：
Chen Nai-hong
中科院分区：
文献类型：
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作者：
Zhu Jie;Chu Shi-feng;Peng Ye;Liu Dan-dan;Chen Chen;Jian Wen-xuan;Sun Hong-shuo;Feng Zhong-ping;Zhang Zhao;Chen Nai-hong
新生儿缺氧缺血性脑损伤（hypoxia-ischemia brain injury，HI）缺乏有效的治疗手段，富含脯氨酸的酪氨酸激酶2（Proline-rich tyrosine kinase 2，Pyk 2）是一种非受体酪氨酸激酶，与成人短暂性缺血性脑损伤高度相关，本研究通过对Pyk 2的研究，发现Pyk 2与HI相关性较强，但Pyk 2与HI相关性较弱。我们研究了Pyk 2在新生儿HI脑损伤中的作用。在出生后7天的小鼠幼仔中，通过单侧颈总动脉结扎，然后缺氧缺血来诱导HI。Pyk 2干扰慢病毒将构建的Pyk 2 shRNA（LV-Pyk 2 shRNA）注射到HI前的新生小鼠一侧脑室，于生后第8- 14天检测脑体积、病理学改变和神经行为学改变。而LV-Pyk 2shRNA注射显著减弱急性HI脑损伤并改善神经行为结果。在氧-葡萄糖剥夺处理的培养皮层神经元中，抑制Pyk 2可显著减轻NMDA受体介导的兴奋性毒性;在新生儿HI脑损伤中也观察到类似的结果。我们证明Pyk 2抑制有助于通过激光散斑对比成像评估的长期脑血管恢复，但在Morris水迷宫和新物体识别测试中，认知功能没有明显改善。因此，我们构建了慢病毒LV-HIF-Pyk 2 shRNA，侧脑室注射LV-HIF-Pyk 2 shRNA可显著改善缺氧环境下大鼠海马神经元的长时程神经元凋亡，并可通过HIF-1α启动子介导的Pyk 2干扰作用，改善缺氧环境下大鼠海马神经元的长时程神经元凋亡。结论：Pyk 2干扰可保护新生鼠脑缺氧缺血损伤，HIF-1α启动子介导的缺氧条件调控可区分缺氧期和正常期，Pyk 2是治疗新生鼠缺氧缺血性脑损伤的一个有前景的药物靶点。
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.