Genetic Deletion or Pharmacological Inhibition of Cyclooxygenase-2 Reduces Blood-Brain Barrier Damage in Experimental Ischemic Stroke
Cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 are two crucial mediators contributing to blood-brain barrier (BBB) damage during cerebral ischemia. However, it is not known whether MMP-9 activation is involved in COX-2-mediated BBB disruption in ischemic stroke. In this study, we hypoth...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-08-01
|
| Series: | Frontiers in Neurology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fneur.2020.00887/full |
| _version_ | 1818195809553874944 |
|---|---|
| author | Changjun Yang Yi Yang Kelly M. DeMars Gary A. Rosenberg Eduardo Candelario-Jalil |
| author_facet | Changjun Yang Yi Yang Kelly M. DeMars Gary A. Rosenberg Eduardo Candelario-Jalil |
| author_sort | Changjun Yang |
| collection | DOAJ |
| description | Cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 are two crucial mediators contributing to blood-brain barrier (BBB) damage during cerebral ischemia. However, it is not known whether MMP-9 activation is involved in COX-2-mediated BBB disruption in ischemic stroke. In this study, we hypothesized that genetic deletion or pharmacological inhibition of COX-2 reduces BBB damage by reducing MMP-9 activity in a mouse model of ischemic stroke. Male COX-2 knockout (COX-2−/−) and wild-type (WT) mice were subjected to 60 min of middle cerebral artery occlusion (MCAO) followed by 24 h of reperfusion. Genetic deletion of COX-2 or post-ischemic treatment with CAY10404, a highly selective COX-2 inhibitor, significantly reduced BBB damage and hemorrhagic transformation, as assessed by immunoglobulin G (IgG) extravasation and brain hemoglobin (Hb) levels, respectively. Immunoblotting analysis showed that tight junction proteins (TJPs) zonula occludens (ZO)-1 and occludin as well as junctional adhesion molecule-A (JAM-A) and the basal lamina protein collagen IV were dramatically reduced in the ischemic brain. Stroke-induced loss of these BBB structural proteins was significantly attenuated in COX-2−/− mice. Similarly, stroke-induced loss of ZO-1 and occludin was significantly attenuated by CAY10404 treatment. Ischemia-induced increase in MMP-9 protein levels in the ipsilateral cerebral cortex was significantly reduced in COX-2−/− mice. Stroke induced a dramatic increase in MMP-9 enzymatic activity in the ischemic cortex, which was markedly reduced by COX-2 gene deficiency or pharmacological inhibition with CAY10404. Levels of myeloperoxidase (MPO, an indicator of neutrophil infiltration into the brain parenchyma), neutrophil elastase (NE), and lipocalin-2 (LCN2, also known as neutrophil gelatinase-associated lipocalin), measured by western blot and specific ELISA kits, respectively, were markedly increased in the ischemic brain. Increased levels of markers for neutrophil infiltration were significantly reduced in COX-2−/− mice compared with WT controls following stroke. Altogether, neurovascular protective effects of COX-2 blockade are associated with reduced BBB damage, MMP-9 expression/activity and neutrophil infiltration. Our study shows for the first time that MMP-9 is an important downstream effector contributing to COX-2-mediated neurovascular damage in ischemic stroke. Targeting the COX-2/MMP-9 pathway could represent a promising strategy to reduce neuroinflammatory events in order to preserve the BBB integrity and ameliorate ischemic stroke injury. |
| first_indexed | 2024-12-12T01:24:05Z |
| format | Article |
| id | doaj.art-ceaff5bac7f24c4f801ac06a918e643c |
| institution | Directory Open Access Journal |
| issn | 1664-2295 |
| language | English |
| last_indexed | 2024-12-12T01:24:05Z |
| publishDate | 2020-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Neurology |
| spelling | doaj.art-ceaff5bac7f24c4f801ac06a918e643c2022-12-22T00:43:09ZengFrontiers Media S.A.Frontiers in Neurology1664-22952020-08-011110.3389/fneur.2020.00887562859Genetic Deletion or Pharmacological Inhibition of Cyclooxygenase-2 Reduces Blood-Brain Barrier Damage in Experimental Ischemic StrokeChangjun Yang0Yi Yang1Kelly M. DeMars2Gary A. Rosenberg3Eduardo Candelario-Jalil4Department of Neuroscience, McKnight Brain Institute, University of Florida, Gainesville, FL, United StatesDepartment of Neurology, Center for Memory and Aging, University of New Mexico Health Sciences Center, Albuquerque, NM, United StatesDepartment of Neuroscience, McKnight Brain Institute, University of Florida, Gainesville, FL, United StatesDepartment of Neurology, Center for Memory and Aging, University of New Mexico Health Sciences Center, Albuquerque, NM, United StatesDepartment of Neuroscience, McKnight Brain Institute, University of Florida, Gainesville, FL, United StatesCyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 are two crucial mediators contributing to blood-brain barrier (BBB) damage during cerebral ischemia. However, it is not known whether MMP-9 activation is involved in COX-2-mediated BBB disruption in ischemic stroke. In this study, we hypothesized that genetic deletion or pharmacological inhibition of COX-2 reduces BBB damage by reducing MMP-9 activity in a mouse model of ischemic stroke. Male COX-2 knockout (COX-2−/−) and wild-type (WT) mice were subjected to 60 min of middle cerebral artery occlusion (MCAO) followed by 24 h of reperfusion. Genetic deletion of COX-2 or post-ischemic treatment with CAY10404, a highly selective COX-2 inhibitor, significantly reduced BBB damage and hemorrhagic transformation, as assessed by immunoglobulin G (IgG) extravasation and brain hemoglobin (Hb) levels, respectively. Immunoblotting analysis showed that tight junction proteins (TJPs) zonula occludens (ZO)-1 and occludin as well as junctional adhesion molecule-A (JAM-A) and the basal lamina protein collagen IV were dramatically reduced in the ischemic brain. Stroke-induced loss of these BBB structural proteins was significantly attenuated in COX-2−/− mice. Similarly, stroke-induced loss of ZO-1 and occludin was significantly attenuated by CAY10404 treatment. Ischemia-induced increase in MMP-9 protein levels in the ipsilateral cerebral cortex was significantly reduced in COX-2−/− mice. Stroke induced a dramatic increase in MMP-9 enzymatic activity in the ischemic cortex, which was markedly reduced by COX-2 gene deficiency or pharmacological inhibition with CAY10404. Levels of myeloperoxidase (MPO, an indicator of neutrophil infiltration into the brain parenchyma), neutrophil elastase (NE), and lipocalin-2 (LCN2, also known as neutrophil gelatinase-associated lipocalin), measured by western blot and specific ELISA kits, respectively, were markedly increased in the ischemic brain. Increased levels of markers for neutrophil infiltration were significantly reduced in COX-2−/− mice compared with WT controls following stroke. Altogether, neurovascular protective effects of COX-2 blockade are associated with reduced BBB damage, MMP-9 expression/activity and neutrophil infiltration. Our study shows for the first time that MMP-9 is an important downstream effector contributing to COX-2-mediated neurovascular damage in ischemic stroke. Targeting the COX-2/MMP-9 pathway could represent a promising strategy to reduce neuroinflammatory events in order to preserve the BBB integrity and ameliorate ischemic stroke injury.https://www.frontiersin.org/article/10.3389/fneur.2020.00887/fullischemic strokeblood-brain barriercyclooxygenase-2matrix metalloproteinase-9tight junction proteins |
| spellingShingle | Changjun Yang Yi Yang Kelly M. DeMars Gary A. Rosenberg Eduardo Candelario-Jalil Genetic Deletion or Pharmacological Inhibition of Cyclooxygenase-2 Reduces Blood-Brain Barrier Damage in Experimental Ischemic Stroke Frontiers in Neurology ischemic stroke blood-brain barrier cyclooxygenase-2 matrix metalloproteinase-9 tight junction proteins |
| title | Genetic Deletion or Pharmacological Inhibition of Cyclooxygenase-2 Reduces Blood-Brain Barrier Damage in Experimental Ischemic Stroke |
| title_full | Genetic Deletion or Pharmacological Inhibition of Cyclooxygenase-2 Reduces Blood-Brain Barrier Damage in Experimental Ischemic Stroke |
| title_fullStr | Genetic Deletion or Pharmacological Inhibition of Cyclooxygenase-2 Reduces Blood-Brain Barrier Damage in Experimental Ischemic Stroke |
| title_full_unstemmed | Genetic Deletion or Pharmacological Inhibition of Cyclooxygenase-2 Reduces Blood-Brain Barrier Damage in Experimental Ischemic Stroke |
| title_short | Genetic Deletion or Pharmacological Inhibition of Cyclooxygenase-2 Reduces Blood-Brain Barrier Damage in Experimental Ischemic Stroke |
| title_sort | genetic deletion or pharmacological inhibition of cyclooxygenase 2 reduces blood brain barrier damage in experimental ischemic stroke |
| topic | ischemic stroke blood-brain barrier cyclooxygenase-2 matrix metalloproteinase-9 tight junction proteins |
| url | https://www.frontiersin.org/article/10.3389/fneur.2020.00887/full |
| work_keys_str_mv | AT changjunyang geneticdeletionorpharmacologicalinhibitionofcyclooxygenase2reducesbloodbrainbarrierdamageinexperimentalischemicstroke AT yiyang geneticdeletionorpharmacologicalinhibitionofcyclooxygenase2reducesbloodbrainbarrierdamageinexperimentalischemicstroke AT kellymdemars geneticdeletionorpharmacologicalinhibitionofcyclooxygenase2reducesbloodbrainbarrierdamageinexperimentalischemicstroke AT garyarosenberg geneticdeletionorpharmacologicalinhibitionofcyclooxygenase2reducesbloodbrainbarrierdamageinexperimentalischemicstroke AT eduardocandelariojalil geneticdeletionorpharmacologicalinhibitionofcyclooxygenase2reducesbloodbrainbarrierdamageinexperimentalischemicstroke |