Rac regulates collagen-induced HSP27 phosphorylation via p44/p42 MAP kinase in human platelets

Kageyama,Yasunari; Doi,Tomoaki; Akamatsu,Shigeru; Kuroyanagi,Gen; Kondo,Akira; Mizutani,Jun; Otsuka,Takanobu; Tokuda,Haruhiko; Kozawa,Osamu; Ogura,Shinji

doi:10.3892/ijmm.2013.1455

Rac regulates collagen-induced HSP27 phosphorylation via p44/p42 MAP kinase in human platelets

Authors:
- Yasunari Kageyama
- Tomoaki Doi
- Shigeru Akamatsu
- Gen Kuroyanagi
- Akira Kondo
- Jun Mizutani
- Takanobu Otsuka
- Haruhiko Tokuda
- Osamu Kozawa
- Shinji Ogura
View Affiliations

Affiliations: Department of Emergency and Disaster Medicine, Gifu University Graduate School of Medicine, Gifu 501-1194, Japan, Department of Intensive Care Medicine, Matsunami General Hospital, Gifu 501-6062, Japan, Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu 501-1194, Japan, Department of Orthopedic Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya 467-8601, Japan, Department of Clinical Laboratory, National Hospital for Geriatric Medicine, National Center for Geriatrics and Gerontology, Obu 474-8511, Japan
Published online on: July 23, 2013 https://doi.org/10.3892/ijmm.2013.1455
Pages: 813-818

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

We previously reported that the collagen-induced phosphorylation of heat shock protein (HSP) 27 via p44/p42 mitogen-activated protein (MAP) kinase is sufficient to induce the secretion of platelet-derived growth factor (PDGF)-AB and the release of soluble CD40 ligand (sCD40L) from human platelets. It has been shown that Rac, which belongs to the Rho family of small GTPases, is involved in the collagen-induced platelet aggregation. In this study, we investigated the role of Rac in the collagen-stimulated release of PDGF-AB and sCD40L in human platelets. Human blood was donated from healthy volunteers and platelet-rich plasma was obtained from the blood samples. The samples were then treated with 1.0 µg/ml collagen for 0, 1, 3, or 5 min and Rac1 activity was determined using the Rac1 Activation Assay kit. We found that collagen stimulated the activation of Rac in human platelets in a time-dependent manner. However, pre-treatment with NSC23766, a selective inhibitor of Rac-guanine nucleotide exchange factor interaction, reduced the collagen-induced platelet aggregation. NSC23766 markedly attenuated not only the collagen-induced p44/p42 MAP kinase phosphorylation, but also the phosphorylation of HSP27 at three serine residues (Ser-15, Ser-78 and Ser-82). In addition, the collagen‑induced release of PDGF-AB and sCD40L was significantly suppressed by NSC23766 in a dose-dependent manner. These results strongly suggest that Rac regulates the collagen-induced HSP27 phosphorylation via p44/p42 MAP kinase in human platelets, resulting in the stimulation of PDGF-AB secretion and the release of sCD40L.

View Figures

View References

1	Kahner BN, Shankar H, Murugappan S, Prasad GL and Kunapuli SP: Nucleotide receptor signaling in platelets. J Thromb Haemost. 4:2317–2326. 2006. View Article : Google Scholar : PubMed/NCBI
2	Li Z, Delaney MK, O’Brien KA and Du X: Signaling during platelet adhesion and activation. Arterioscler Thromb Vasc Biol. 30:2341–2349. 2010. View Article : Google Scholar : PubMed/NCBI
3	Benjamin IJ and McMillan DR: Stress (heat shock) proteins molecular chaperones in cardiovascular biology and disease. Circ Res. 83:117–132. 1998. View Article : Google Scholar : PubMed/NCBI
4	Mymrikov EV, Seit-Nebi AS and Gusev NB: Large potentials od small heat shock proteins. Physiol Rev. 91:1123–1159. 2011. View Article : Google Scholar : PubMed/NCBI
5	Kahn ML: Platelet-collagen responses: molecular basis and therapeutic promise. Semin Thromb Hemost. 30:419–425. 2004. View Article : Google Scholar : PubMed/NCBI
6	Shattil SJ and Newman PJ: Integrins: dynamic scaffolds for adhesion and signaling in platelets. Blood. 104:1606–1615. 2004. View Article : Google Scholar : PubMed/NCBI
7	Nieswandt B and Watson SP: Platelet-collagen interaction: is GPVI the central receptor? Blood. 102:449–461. 2003. View Article : Google Scholar : PubMed/NCBI
8	Moroi M and Jung SM: Platelet grycoprotein VI: its structure and function. Thromb Res. 114:221–233. 2004. View Article : Google Scholar : PubMed/NCBI
9	Saklatvala J, Rawlinson L, Waller RJ, Sarsfield S, Lee JC, Morton LF, Barnes MJ and Farndale RW: Role for p38 mitogen-activated protein kinase in platelet aggregation caused by collagen or a thromboxane analogue. J Biol Chem. 271:6586–6589. 1996. View Article : Google Scholar : PubMed/NCBI
10	Kato H, Adachi S, Doi T, Matsushima-Nishiwaki R, Minamitani C, Akamatsu S, Enomoto Y, Tokuda H, Otsuka T, Iwama T, Kozawa O and Ogura S: Mechanism of collagen-induced release of 5-HT, PDGF-AB and sCD40L from human platelets: role of HSP27 phosphorylation via p44/p42 MAPK. Thromb Res. 126:39–43. 2010. View Article : Google Scholar : PubMed/NCBI
11	Takai Y, Sasaki T and Matozaki T: Small GTP-binding proteins. Physiol Rev. 81:153–208. 2001.PubMed/NCBI
12	Pleines I, Elvers M, Strehl A, Pozgajova M, Varga-Szabo D, May F, Chrostek-Grashoff A, Brakebusch C and Nieswandt B: Rac1 is essential for phospholipase C-γ2 activation in platelets. Pflugers Arch. 457:1173–1185. 2009.
13	Soulet C, Gendreau S, Missy K, Benard V, Plantavid M and Payrastre B: Characterisation of Rac activation in thrombin-and collagen-stimulated human blood platelets. FEBS Lett. 507:253–258. 2001. View Article : Google Scholar : PubMed/NCBI
14	Tohgi H, Takahashi H, Watanabe K, Kuki H and Shirasawa Y: Development of large platelet aggregates from small aggregates as determined by laser-light scattering: effects of aggregant concentration and antiplatelet medication. Thromb Haemost. 75:838–843. 1996.
15	Kato K, Ito H, Hasegawa K, Inaguma Y, Kozawa O and Asano T: Modulation of the stress-induced synthesis of hsp27 and αB-crystallin by cyclic AMP in C6 rat glioma cells. J Neurochem. 66:946–950. 1996.
16	Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar : PubMed/NCBI
17	Gao Y, Dickerson JB, Guo F, Zheng J and Zheng Y: Rational design and characterization of a Rac GTPase-specific small molecule inhitor. Proc Natl Acad Sci USA. 101:7618–7623. 2004. View Article : Google Scholar : PubMed/NCBI
18	Davi G and Patrono C: Platelet activation and atherothrombosis. N Engl J Med. 357:2482–2494. 2007. View Article : Google Scholar : PubMed/NCBI
19	Rendu F and Brohard-Bohn B: The platelet release reaction: granules’ constituents, secretion and functions. Platelets. 12:261–273. 2001.
20	Heldin CH and Westermark B: Mechanism of action and in vivo role of platelet-derived growth factor. Physiol Rev. 79:1283–1316. 1999.PubMed/NCBI
21	Hermann A, Rauch BH, Braun M, Schror K and Weber AA: Platelet CD40 ligand (CD40L)-subcellular localization, regulation of expression, and inhibition by clopidogrel. Platelets. 12:74–82. 2001. View Article : Google Scholar : PubMed/NCBI
22	Andre P, Nannizzi-Alaimo L, Prasad SK and Phillips DR: Platelet-derived CD40L: the switch-hitting player of cardiovascular disease. Circulation. 106:896–899. 2002. View Article : Google Scholar : PubMed/NCBI
23	Henn V, Slupsky JR, Grafe M, Anagnostopoulos I, Forster R, Muller-Berghaus G and Kroczek RA: CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells. Nature. 391:591–594. 1998. View Article : Google Scholar : PubMed/NCBI
24	Heeschen C, Dimmeler S, Hamm CW, van den Brand MJ, Boersma E, Zeiher AM and Simoons ML; CAPTURE Study Investigators. Soluble CD40 ligand in acute coronary syndromes. N Engl J Med. 348:1104–1111. 2003. View Article : Google Scholar : PubMed/NCBI