Blockade of HCN2 Channels Provides Neuroprotection Against Ischemic Injury via Accelerating Autophagic Degradation in Hippocampal Neurons

• 作者: Chang-Jun Li Cheng Chen Li Liu Lian-Jun Guo Ping Jing Xian-Gang Zong Xiao-Xue Zhang Ya-Qiao Shu Yun Lu
• 来源期刊: 神经科学通报（英文版）
• 年，卷（期）: 2020,（8）
• 所属期刊栏目: Original Articles
• 页码范围: 875-894
• 页数: 20页
• 语种: 英文

摘要

In the central nervous system,hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are essential to maintain normal neuronal function.Recent studies have shown that HCN channels may be involved in the pathological process of ischemic brain injury,but the mechanisms remain unclear.Autophagy is activated in cerebral ischemia,but its role in cell death/survival remains controversial.In this study,our results showed that the HCN channel blocker ZD7288 remarkably decreased the percentage of apoptotic neurons and corrected the excessive autophagy induced by oxygen-glucose deprivation followed by reperfusion (OGD/R) in hippocampal HT22 neurons.Furthermore,in the OGD/R group,p-mTOR,p-ULK1 (Ser757),and p62 were significantly decreased,while p-ULK1 (Ser317),atg5,and beclinl were remarkably increased.ZD7288 did not change the expression of p-ULK1 (Ser757),ULK1 (Ser317),p62,Beclinl,and atg5,which are involved in regulating autophagosome formation.Besides,we found that OGD/R induced a significant increase in Cathepsin D expression,but not LAMP-1.Treatment with ZD7288 at 10 μrnol/L in the OGD/R group did not change the expression of cathepsin D and LAMP-1.However,chloroquine (CQ),which decreases autophagosome-lysosome fusion,eliminated the correction of excessive autophagy and neuroprotection by ZD7288.Besides,shRNA knockdown of HCN2 channels significantly reduced the accumulation of LC3-Ⅱ and increased neuron survival in the OGD/R and transient global cerebral ischemia (TGCI) models,and CQ also eliminated the effects of HCN2-shRNA.Furthermore,we found that the percentage of LC3-positive puncta that co-localized with LAMP-l-positive lysosomes decreased in Con-shRNA-transfected HT22 neurons exposed to OGD/R or CQ.In HCN2-shRNA-transfected HT22 neurons,the percentage of LC3-positive puncta that co-localized with LAMP-l-positive lysosomes increased under OGD/R;however,the percentage was significantly decreased by the addition of CQ to HCN2-shRNA-transfected HT22 neurons.The present results demonstrated that blockade of HCN2 channels provides neuroprotection against OGD/R and TGCI by accelerating autophagic degradation attributable to the promotion of autophagosome and lysosome fusion.

作者信息

序号	姓名	单位	发文数	被引次数	H指数	G指数
1	Cheng Chen		1	0	0.0	0.0
2	Li Liu		1	0	0.0	0.0
3	Ya-Qiao Shu		1	0	0.0	0.0
4	Ping Jing		1	0	0.0	0.0
5	Yun Lu		1	0	0.0	0.0
6	Xiao-Xue Zhang		1	0	0.0	0.0
7	Xian-Gang Zong		1	0	0.0	0.0
8	Lian-Jun Guo		1	0	0.0	0.0
9	Chang-Jun Li		1	0	0.0	0.0