Vascular superoxide production by endothelin-1 requires Src non-receptor protein tyrosine kinase and MAPK activation.
ET-1 induces vascular O(2)(*-) production via activation of NADPH oxidase. We have investigated whether c-Src and MAPKs activation are involved in ET-1-induced vascular oxidative response. At 2 h, ET-1 induced an increase in NADPH oxidase-driven O(2)(*-) production in rat isolated aortic rings, which was completely suppressed in PP2 (c-Src inhibitor)-pretreated rings, whereas PP3 (inactive analogue of PP2) was without effect. ET-1 increased the levels of phospho-c-Src, the active form of c-Src, and the phosphorylation of cortactin, a Src-specific substrate. Both c-Src and cortactin phosphorylation induced by ET-1 were prevented by PP2. The increased expression of p47(phox), the main cytosolic subunit of NADPH oxidase, induced by ET-1 was also prevented by PP2. The increased vascular O(2)(*-) production and p47(phox) up-regulation induced by ET-1 was only inhibited in aortic rings coincubated with the ERK1/2 inhibitor, PD98059; being without effects both the p38 MAPK inhibitor, SB203580, and JNK inhibitor, SP600125. Aortic rings incubation with ET-1 increased the phosphorylation of ERK1/2. This effect was suppressed by coincubation with PP2 showing that this event is down-stream of c-Src activation. In conclusion, ET-1 induces NADPH oxidase-driven O(2)(*-) generation through increase of p47(phox) protein expression. The signalling pathway for this effect involves c-Src activation and ERK1/2 phosphorylation.
Romero
Miguel
M
Department of Pharmacology, School of Pharmacy, University of Granada, 18071 Granada, Spain.
Jiménez
Rosario
R
Sánchez
Manuel
M
López-Sepúlveda
Rocío
R
Zarzuelo
Antonio
A
Tamargo
Juan
J
Pérez-Vizcaíno
Francisco
F
Duarte
Juan
J
D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000201:enzymology*; D000081247:CSK Tyrosine-Protein Kinase; D051356:Cortactin / Q000378:metabolism; D019332:Endothelin-1 / Q000378:metabolism*; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D008297:Male; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D013481:Superoxides / Q000378:metabolism*; D013997:Time Factors; D019061:src-Family Kinases
D051356:Cortactin; C495870:Cttn protein, rat; D019332:Endothelin-1; D047428:Protein Kinase Inhibitors; D013481:Superoxides; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; D011505:Protein-Tyrosine Kinases; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases