Nitric Oxide Interacts with Caveolin-1 to Facilitate Autophagy-Lysosome-Mediated Claudin-5 Degradation in Oxygen-Glucose Deprivation-Treated Endothelial Cells
机构：[1]Translational Center for Stem Cell Research, Tongji Hospital, Stem Cell Research Center, Tongji University School of Medicine, Shanghai 200065, China[2]The Central Laboratory, Shenzhen Second People’s Hospital, Shenzhen University 1st Affiliated Hospital, Shenzhen 518035, China[3]Department of Neurosurgery and Shenzhen Key Laboratory of Neurosurgery, Shenzhen Second People’s Hospital, Shenzhen University 1st Affiliated Hospital, Shenzhen 518035, China[4]Department of Pharmaceutical Sciences, College of Pharmacy, University of New Mexico, Albuquerque, NM 87131-0001, USA[5]Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, the Second Affiliated Hospital of Soochow University, Soochow University, Suzhou 215004, China
Using in vitro oxygen-glucose deprivation (OGD) model, we have previously demonstrated that 2-h OGD induces rapid, caveolin-1-mediated dissociation of claudin-5 from the cellular cytoskeletal framework and quick endothelial barrier disruption. In this study, we further investigated the fate of translocated claudin-5 and the mechanisms by which OGD promotes caveolin-1 translocation. Exposure of bEND3 cells to 4-h OGD, but not 2-h OGD plus 2-h reoxygenation, resulted in claudin-5 degradation. Inhibition of autophagy or the fusion of autophagosome with lysosome, but not proteasome, blocked OGD-induced claudin-5 degradation. Moreover, knockdown of caveolin-1 with siRNA blocked OGD-induced claudin-5 degradation. Western blot analysis showed a transient colocalization of caveolin-1, claudin-5, and LC3B in autolysosome or lipid raft fractions at 2-h OGD. Of note, inhibiting autophagosome and lysosome fusion sustained the colocalization of caveolin-1, claudin-5, and LC3B throughout the 4-h OGD exposure. EPR spin trapping showed increased nitric oxide (NO) generation in 2-h OGD-treated cells, and inhibiting NO with its scavenger C-PTIO or inducible nitric oxide synthase (iNOS) inhibitor 1400W prevented OGD-induced caveolin-1 translocation and claudin-5 degradation. Taken together, our data provide a novel mechanism underlying endothelial barrier disruption under prolonged ischemic conditions, in which NO promotes caveolin-1-mediated delivery of claudin-5 to the autophagosome for autophagy-lysosome-dependent degradation.
基金：
This work was supported by grants from the National Natural
Science Foundation of China (81371328), Shenzhen Science and
Technology Innovation Commission (CXZZ20130516152706040 and
ZDSY20140509173142601), and the National Institutes of Health
(P20RR15636).
第一作者机构：[1]Translational Center for Stem Cell Research, Tongji Hospital, Stem Cell Research Center, Tongji University School of Medicine, Shanghai 200065, China
通讯作者：
通讯机构：[2]The Central Laboratory, Shenzhen Second People’s Hospital, Shenzhen University 1st Affiliated Hospital, Shenzhen 518035, China[3]Department of Neurosurgery and Shenzhen Key Laboratory of Neurosurgery, Shenzhen Second People’s Hospital, Shenzhen University 1st Affiliated Hospital, Shenzhen 518035, China[4]Department of Pharmaceutical Sciences, College of Pharmacy, University of New Mexico, Albuquerque, NM 87131-0001, USA
推荐引用方式(GB/T 7714)：
Jie Liu,John Weaver,Xinchun Jin,et al.Nitric Oxide Interacts with Caveolin-1 to Facilitate Autophagy-Lysosome-Mediated Claudin-5 Degradation in Oxygen-Glucose Deprivation-Treated Endothelial Cells[J].MOLECULAR NEUROBIOLOGY.2016,53(9):5935-5947.doi:10.1007/s12035-015-9504-8.
APA：
Jie Liu,John Weaver,Xinchun Jin,Yuan Zhang,Ji Xu...&Wenlan Liu.(2016).Nitric Oxide Interacts with Caveolin-1 to Facilitate Autophagy-Lysosome-Mediated Claudin-5 Degradation in Oxygen-Glucose Deprivation-Treated Endothelial Cells.MOLECULAR NEUROBIOLOGY,53,(9)
MLA：
Jie Liu,et al."Nitric Oxide Interacts with Caveolin-1 to Facilitate Autophagy-Lysosome-Mediated Claudin-5 Degradation in Oxygen-Glucose Deprivation-Treated Endothelial Cells".MOLECULAR NEUROBIOLOGY 53..9(2016):5935-5947
