p38 Mitogen-Activated Protein Kinase Mediates Hyperosmolarity-Induced Vasoconstriction through Myosin Light Chain Phosphorylation and Actin Polymerization in Rat Aorta

• Published in: Biological & pharmaceutical bulletin Vol. 36; no. 11; pp. 1849 - 1856
• Main Authors: Sasahara, Tomoya, Yayama, Katsutoshi, Okamoto, Hiroshi
• Format: Journal Article
• Language: English
• Published: Japan The Pharmaceutical Society of Japan 01-11-2013; Pharmaceutical Society of Japan; Japan Science and Technology Agency
• Subjects: actin polymerization; Actins - physiology; Animals; Aorta, Thoracic - drug effects; Aorta, Thoracic - physiology; Calcium - physiology; hyperosmolarity; In Vitro Techniques; Male; Myocytes, Smooth Muscle - physiology; Myosin Light Chains - physiology; Osmolar Concentration; Osmosis; Osmotic Pressure - physiology; p38 mitogen-activated protein kinase; p38 Mitogen-Activated Protein Kinases - physiology; Phosphorylation - drug effects; Polymerization; Protein Phosphatase 1 - physiology; Rats; Rats, Wistar; Rho-kinase; Sucrose - pharmacology; vasoconstriction; Vasoconstriction - drug effects; Vasoconstriction - physiology
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.b13-00563

Hyperosmotic stress induces the contractile response of vascular smooth muscle cells (VSMCs). Previous studies have demonstrated that cytoskeleton reorganization and Rho/Rho-kinase-mediated inactivation of myosin light chain phosphatase (MLCP) play an important role in hyperosmotic vasoconstriction, but the precise mechanism is unknown. This study aimed to investigate the contractile response of endothelium-denuded rings of rat aortas to hyperosmolar sucrose (160 m M ) in the presence or absence of inhibitors for various protein kinases. We found that the hyperosmotic constriction of aortic rings was attenuated not only by ML-7 or hydroxyfasudil, specific inhibitor for myosin light chain kinase (MLCK) or Rho-kinase, respectively, but also by SB203580, a specific inhibitor for p38 mitogen-activated kinase (p38 MAPK). Hyperosmolar sucrose evoked a transient increase in cytosolic free Ca2+ in rat VSMCs, and this response was not affected by SB203580. Western blot analysis of proteins extracted from rings showed that the hyperosmolar sucrose stimulated phosphorylation of the Rho-kinase-mediated myosin phosphatase target subunit 1, myosin light chain (MLC), and p38 MAPK. The experiments performed using a combination of the kinase inhibitors showed that hyperosmolarity-induced MLC phosphorylation is partially mediated via the SB203580-sensitive pathway and is independent of both MLCK and Rho-kinase-mediated inactivation of MLCP. Furthermore, the hyperosmolarity-induced increase in the F-actin/G-actin ratio in rings was attenuated not only by hydroxyfasudil but also by SB203580. These results suggest that p38 MAPK is involved in hyperosmotic vasoconstriction via stimulation of MLC phosphorylation and cytoskeleton reorganization through pathways independent of activation of MLCK and/or Rho-kinase-mediated mechanisms.