He Zhi, Chu Xiao-Jiao, Zeng Xiao-Li, Zhang Xiao-Xue, Chen Jing, Wang Ke, Chen Yue, Li Zi-Cheng, Zhao Bo
Department of Pharmacology, College of Medicine, Jiaxing University, Jiaxing, 314001, People's Republic of China.
Third-Grade Pharmacological Laboratory on Traditional Chinese Medicine, State Administration of Traditional Chinese Medicine, China Three Gorges University, Yichang, 443002, People's Republic of China.
Sci Rep. 2025 May 25;15(1):18255. doi: 10.1038/s41598-025-03172-8.
To study the effect of inhibition of hyperpolarization-activated cyclic nucleotide-gated cation channel (HCN) on spatial memory with cerebral ischemia-reperfusion (IR) in rats, the middle cerebral artery occlusion was performed to induce the IR injury. ZD7288 (5 µg/µl) and MK-801 (100 µg/µl) were administrated by lateral ventricular injection. Except for the sham group, all other groups were subjected to ischemia for 2 h followed by reperfusion for 4 weeks. The neurological impairment was evaluated using the neurological deficit score, and the Morris water maze detected changes in spatial memory. The cerebral infarction volume was determined by TTC staining, and the morphology and number of hippocampal neurons on the ischemic side were observed through HE staining. Immunoblotting and qPCR were used to detect the expressions of HCN1, HCN2, and N-methyl-D-aspartic acid receptor (NMDAR) subunits in the hippocampus. The expression changes of NMDAR subunits were applied by immunohistochemistry in different hippocampus regions. Our study found that ZD7288 could improve the spatial memory deficits induced by IR and exert neuroprotective effects. The possible mechanism is related to the fact that ZD7288 regulates the expression of various subunits of NMDAR by inhibiting HCN, including its effect on their phosphorylation states.
为研究抑制超极化激活的环核苷酸门控阳离子通道(HCN)对大鼠脑缺血再灌注(IR)后空间记忆的影响,采用大脑中动脉闭塞法诱导IR损伤。通过侧脑室注射给予ZD7288(5µg/µl)和MK-801(100µg/µl)。除假手术组外,其他所有组均缺血2小时,然后再灌注4周。使用神经功能缺损评分评估神经功能障碍,通过Morris水迷宫检测空间记忆的变化。通过TTC染色测定脑梗死体积,通过HE染色观察缺血侧海马神经元的形态和数量。采用免疫印迹和qPCR检测海马中HCN1、HCN2和N-甲基-D-天冬氨酸受体(NMDAR)亚基的表达。通过免疫组织化学检测不同海马区域NMDAR亚基的表达变化。我们的研究发现,ZD7288可以改善IR诱导的空间记忆缺陷并发挥神经保护作用。可能的机制与ZD7288通过抑制HCN调节NMDAR各亚基的表达有关,包括其对它们磷酸化状态的影响。