机构地区:[1]Department of Pharmacology,School of Basic Medical Sciences,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China [2]Department of Neurology,The Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China [3]Office of Academic Research,The Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China [4]Department of Pharmacy,Zhongnan Hospital of Wuhan University,Wuhan 430030,China [5]Department of Clinical Laboratory,Wuhan PuAi Hospital,Wuhan 430033,China [6]Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation,Wuhan 430030,China [7]Center for Integrated Protein Science and Zentrum fur Pharmaforschung,Department Pharmazie,Ludwig-Maximilians-Universitat Miinchen,80539 Munich,Germany
出 处:《Neuroscience Bulletin》2020年第8期875-894,共20页神经科学通报(英文版)
基 金:supported by grants from the National Natural Science Foundation of China (85100929);the Natural Science Foundation of Hubei Province,China (2018CFB302 and 2019CFB446);the Youth Fund of Health and Family Planning Commission of Wuhan Municipality,Hubei Province,China (WX18Q13 and WX18Q22)。
摘 要:In the central nervous system,hyperpolarizationactivated 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 beclin1 were remarkably increased.ZD7288 did not change the expression of p-ULK1(Ser757),ULK1(Ser317),p62,Beclin1,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 10μmol/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-Ⅱand 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-1-positive lysosomes decreased in Con-shRNAtransfected 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-1-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 chan
关 键 词:HCN2 channel AUTOPHAGY NEUROPROTECTION Oxygen-glucose deprivation/reperfusion Transient global cerebral ischemia
分 类 号:R743.3[医药卫生—神经病学与精神病学]
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...