p32-Dependent p38 MAPK Activation by Arginase II Downregulation Contributes to Endothelial Nitric Oxide Synthase Activation in HUVECs
Arginase II reciprocally regulates endothelial nitric oxide synthase (eNOS) through a p32-dependent Ca<sup>2+</sup> control. We investigated the signaling pathway of arginase II-dependent eNOS phosphorylation. Western blot analysis was applied for examining protein activation and [Ca<...
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MDPI AG
2020-02-01
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| Online Access: | https://www.mdpi.com/2073-4409/9/2/392 |
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| author | Bon-Hyeock Koo Moo-Ho Won Young-Myeong Kim Sungwoo Ryoo |
| author_facet | Bon-Hyeock Koo Moo-Ho Won Young-Myeong Kim Sungwoo Ryoo |
| author_sort | Bon-Hyeock Koo |
| collection | DOAJ |
| description | Arginase II reciprocally regulates endothelial nitric oxide synthase (eNOS) through a p32-dependent Ca<sup>2+</sup> control. We investigated the signaling pathway of arginase II-dependent eNOS phosphorylation. Western blot analysis was applied for examining protein activation and [Ca<sup>2+</sup>]c was analyzed by microscopic and FACS analyses. Nitric oxide (NO) and reactive oxygen species (ROS) productions were measured using specific fluorescent dyes under microscopy. NO signaling pathway was tested by measuring vascular tension. Following arginase II downregulation by chemical inhibition or gene knockout (KO, ArgII<sup>−/−</sup>), increased eNOS phosphorylation at Ser1177 and decreased phosphorylation at Thr495 was depend on p38 MAPK activation, which induced by CaMKII activation through p32-dependent increase in [Ca<sup>2+</sup>]c. The protein amount of p32 negatively regulated p38 MAPK activation. p38 MAPK contributed to Akt-induced eNOS phosphorylation at Ser1177 that resulted in accelerated NO production and reduced reactive oxygen species production in aortic endothelia. In vascular tension assay, p38 MAPK inhibitor decreased acetylcholine-induced vasorelaxation responses and increased phenylephrine-dependent vasoconstrictive responses. In ApoE<sup>−/−</sup> mice fed a high cholesterol diet, arginase II inhibition restored p32/CaMKII/p38 MAPK/Akt/eNOS signaling cascade that was attenuated by p38 MAPK inhibition. Here, we demonstrated a novel signaling pathway contributing to understanding of the relationship between arginase II, endothelial dysfunction, and atherogenesis. |
| first_indexed | 2024-03-12T06:40:27Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T06:40:27Z |
| publishDate | 2020-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-bb509a3e44964610beb5b0fa3f26e0c72023-09-03T01:05:54ZengMDPI AGCells2073-44092020-02-019239210.3390/cells9020392cells9020392p32-Dependent p38 MAPK Activation by Arginase II Downregulation Contributes to Endothelial Nitric Oxide Synthase Activation in HUVECsBon-Hyeock Koo0Moo-Ho Won1Young-Myeong Kim2Sungwoo Ryoo3Department of Biological Sciences, Kangwon National University, Chuncheon, Gangwon 24341, KoreaDepartment of Neurobiology, School of Medicine, Kangwon National University, Chuncheon, Gangwon 24341, KoreaDepartment of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon 24341, KoreaDepartment of Biological Sciences, Kangwon National University, Chuncheon, Gangwon 24341, KoreaArginase II reciprocally regulates endothelial nitric oxide synthase (eNOS) through a p32-dependent Ca<sup>2+</sup> control. We investigated the signaling pathway of arginase II-dependent eNOS phosphorylation. Western blot analysis was applied for examining protein activation and [Ca<sup>2+</sup>]c was analyzed by microscopic and FACS analyses. Nitric oxide (NO) and reactive oxygen species (ROS) productions were measured using specific fluorescent dyes under microscopy. NO signaling pathway was tested by measuring vascular tension. Following arginase II downregulation by chemical inhibition or gene knockout (KO, ArgII<sup>−/−</sup>), increased eNOS phosphorylation at Ser1177 and decreased phosphorylation at Thr495 was depend on p38 MAPK activation, which induced by CaMKII activation through p32-dependent increase in [Ca<sup>2+</sup>]c. The protein amount of p32 negatively regulated p38 MAPK activation. p38 MAPK contributed to Akt-induced eNOS phosphorylation at Ser1177 that resulted in accelerated NO production and reduced reactive oxygen species production in aortic endothelia. In vascular tension assay, p38 MAPK inhibitor decreased acetylcholine-induced vasorelaxation responses and increased phenylephrine-dependent vasoconstrictive responses. In ApoE<sup>−/−</sup> mice fed a high cholesterol diet, arginase II inhibition restored p32/CaMKII/p38 MAPK/Akt/eNOS signaling cascade that was attenuated by p38 MAPK inhibition. Here, we demonstrated a novel signaling pathway contributing to understanding of the relationship between arginase II, endothelial dysfunction, and atherogenesis.https://www.mdpi.com/2073-4409/9/2/392arginase iicalciumendothelial nitric oxide synthasep38 mapkp32 |
| spellingShingle | Bon-Hyeock Koo Moo-Ho Won Young-Myeong Kim Sungwoo Ryoo p32-Dependent p38 MAPK Activation by Arginase II Downregulation Contributes to Endothelial Nitric Oxide Synthase Activation in HUVECs Cells arginase ii calcium endothelial nitric oxide synthase p38 mapk p32 |
| title | p32-Dependent p38 MAPK Activation by Arginase II Downregulation Contributes to Endothelial Nitric Oxide Synthase Activation in HUVECs |
| title_full | p32-Dependent p38 MAPK Activation by Arginase II Downregulation Contributes to Endothelial Nitric Oxide Synthase Activation in HUVECs |
| title_fullStr | p32-Dependent p38 MAPK Activation by Arginase II Downregulation Contributes to Endothelial Nitric Oxide Synthase Activation in HUVECs |
| title_full_unstemmed | p32-Dependent p38 MAPK Activation by Arginase II Downregulation Contributes to Endothelial Nitric Oxide Synthase Activation in HUVECs |
| title_short | p32-Dependent p38 MAPK Activation by Arginase II Downregulation Contributes to Endothelial Nitric Oxide Synthase Activation in HUVECs |
| title_sort | p32 dependent p38 mapk activation by arginase ii downregulation contributes to endothelial nitric oxide synthase activation in huvecs |
| topic | arginase ii calcium endothelial nitric oxide synthase p38 mapk p32 |
| url | https://www.mdpi.com/2073-4409/9/2/392 |
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