Prostaglandin E2 dependent migration of human brain endothelial cells is mediated through Rho-Kinase-II.
Prostaglandin E2 (PGE2), that plays a crucial role in angiogenesis as well as in ischemic and inflammatory disorders of the brain, is associated with breakdown of the blood-brain barrier (BBB). Previously, we had shown that PGE2-induced human brain endothelial cells (HBECs) migration, and works in a...
Saved in:
| Main Authors: | , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2025-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0326312 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849245783300243456 |
|---|---|
| author | Gausal Azam Khan Arjun Ghosh |
| author_facet | Gausal Azam Khan Arjun Ghosh |
| author_sort | Gausal Azam Khan |
| collection | DOAJ |
| description | Prostaglandin E2 (PGE2), that plays a crucial role in angiogenesis as well as in ischemic and inflammatory disorders of the brain, is associated with breakdown of the blood-brain barrier (BBB). Previously, we had shown that PGE2-induced human brain endothelial cells (HBECs) migration, and works in a cooperative manner through its three receptors (EP2, EP3 & EP4). However, the detailed signaling mechanism of PGE2-induced HBECs migration remains obscure. In this present study, we investigated the signaling pathway of actin dynamics/polymerization and migration of HBECs by PGE2 in vitro. Expression of ROCK was analyzed by ELISA and RT-PCR. Actin polymerization was evaluated by NBD-phallacidin immunofluorescence staining. HBECs expressed only ROCK II. PGE2 (100 pM) induced ROCK II expression occurs in dose-and-time-dependent manner. ROCK II inhibition by Y27632 (150nM), as well as ROCK II silencing significantly attenuated PGE2-induced migration of HBECs. We further showed that pretreatment of PKA inhibitor (H-89; 0.5 μM) or adenylate cyclase inhibitor (ddA; 1μM) completely inhibited PGE2-induced ROCK II activity. Furthermore, PGE2-induced MLC phosphorylation also occurs in a time-dependent manner. However, pretreatment of ROCK II inhibitor or silencing of ROCK II significantly abrogated PGE2-induced MLC phosphorylation as well as F-actin polymerization. Our ex-vivo aortic ring angiogenesis study also showed that pretreatment of ROCK II inhibitor significantly inhibited ECs sprouting. These results suggest that PGE2-induced HBECs migration is mediated through PKA, ROCK II and MLC phosphorylation as well as F-actin polymerization, indicating that modulation of these pathways may aid in the future treatment of dysregulated angiogenesis in cerebrovascular diseases. |
| format | Article |
| id | doaj-art-3ca0df689bce4aa2bc48991aeaaa3432 |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-3ca0df689bce4aa2bc48991aeaaa34322025-08-20T03:58:41ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01206e032631210.1371/journal.pone.0326312Prostaglandin E2 dependent migration of human brain endothelial cells is mediated through Rho-Kinase-II.Gausal Azam KhanArjun GhoshProstaglandin E2 (PGE2), that plays a crucial role in angiogenesis as well as in ischemic and inflammatory disorders of the brain, is associated with breakdown of the blood-brain barrier (BBB). Previously, we had shown that PGE2-induced human brain endothelial cells (HBECs) migration, and works in a cooperative manner through its three receptors (EP2, EP3 & EP4). However, the detailed signaling mechanism of PGE2-induced HBECs migration remains obscure. In this present study, we investigated the signaling pathway of actin dynamics/polymerization and migration of HBECs by PGE2 in vitro. Expression of ROCK was analyzed by ELISA and RT-PCR. Actin polymerization was evaluated by NBD-phallacidin immunofluorescence staining. HBECs expressed only ROCK II. PGE2 (100 pM) induced ROCK II expression occurs in dose-and-time-dependent manner. ROCK II inhibition by Y27632 (150nM), as well as ROCK II silencing significantly attenuated PGE2-induced migration of HBECs. We further showed that pretreatment of PKA inhibitor (H-89; 0.5 μM) or adenylate cyclase inhibitor (ddA; 1μM) completely inhibited PGE2-induced ROCK II activity. Furthermore, PGE2-induced MLC phosphorylation also occurs in a time-dependent manner. However, pretreatment of ROCK II inhibitor or silencing of ROCK II significantly abrogated PGE2-induced MLC phosphorylation as well as F-actin polymerization. Our ex-vivo aortic ring angiogenesis study also showed that pretreatment of ROCK II inhibitor significantly inhibited ECs sprouting. These results suggest that PGE2-induced HBECs migration is mediated through PKA, ROCK II and MLC phosphorylation as well as F-actin polymerization, indicating that modulation of these pathways may aid in the future treatment of dysregulated angiogenesis in cerebrovascular diseases.https://doi.org/10.1371/journal.pone.0326312 |
| spellingShingle | Gausal Azam Khan Arjun Ghosh Prostaglandin E2 dependent migration of human brain endothelial cells is mediated through Rho-Kinase-II. PLoS ONE |
| title | Prostaglandin E2 dependent migration of human brain endothelial cells is mediated through Rho-Kinase-II. |
| title_full | Prostaglandin E2 dependent migration of human brain endothelial cells is mediated through Rho-Kinase-II. |
| title_fullStr | Prostaglandin E2 dependent migration of human brain endothelial cells is mediated through Rho-Kinase-II. |
| title_full_unstemmed | Prostaglandin E2 dependent migration of human brain endothelial cells is mediated through Rho-Kinase-II. |
| title_short | Prostaglandin E2 dependent migration of human brain endothelial cells is mediated through Rho-Kinase-II. |
| title_sort | prostaglandin e2 dependent migration of human brain endothelial cells is mediated through rho kinase ii |
| url | https://doi.org/10.1371/journal.pone.0326312 |
| work_keys_str_mv | AT gausalazamkhan prostaglandine2dependentmigrationofhumanbrainendothelialcellsismediatedthroughrhokinaseii AT arjunghosh prostaglandine2dependentmigrationofhumanbrainendothelialcellsismediatedthroughrhokinaseii |