Significant correlation between the acceleration of platelet aggregation and phosphorylation of HSP27 at Ser-78 in diabetic patients

Tokuda,Haruhiko; Kato,Kenji; Kasahara,Senji; Matsushima-Nishiwaki,Rie; Mizuno,Takahiko; Sakakibara,Seiko; Kozawa,Osamu

doi:10.3892/ijmm.2012.1146

Significant correlation between the acceleration of platelet aggregation and phosphorylation of HSP27 at Ser-78 in diabetic patients

Authors:
- Haruhiko Tokuda
- Kenji Kato
- Senji Kasahara
- Rie Matsushima-Nishiwaki
- Takahiko Mizuno
- Seiko Sakakibara
- Osamu Kozawa
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Affiliations: Department of Clinical Laboratory, National Center for Geriatrics and Gerontology, Obu, Aichi 474-8511, Japan, Department of Pharmacology, Gifu University, Graduate School of Medicine, Gifu 501-1194, Japan, First Department of Internal Medicine, Gifu University, Graduate School of Medicine, Gifu 501-1194, Japan
Published online on: October 1, 2012 https://doi.org/10.3892/ijmm.2012.1146
Pages: 1387-1395

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Abstract

To clarify the mechanism underlying a high risk of thrombotic complications in diabetic patients, we investigated the relationship between HSP27 phosphorylation and the platelet activation induced by adenosine diphosphate (ADP) in diabetic patients. Platelet-rich plasma was prepared from the blood of type 2 diabetes mellitus (DM) patients. By measuring the dose response of platelet aggregation to ADP, an individual ED50 was determined. Based on the normal range identified in non-DM controls, the subjects were divided into a hyper-aggregate (Group 1) and a normo- or hypo-aggregate group (Group 2). The protein phosphorylation was analyzed by western blotting. The release of PDGF-AB and sCD40 ligand (sCD40L) was measured by ELISA. In both groups, ADP induced HSP27 phosphorylation at Ser-78 and Ser-82. The phosphorylation at Ser-78 and the release of both PDGF-AB and sCD40L induced by a low dose of ADP (1 µM) in Group 1 were significantly higher than these values in Group 2. There was a significant relationship between the ADP-induced HSP27 phosphorylation level at Ser-78 and the ADP ED50 value of platelet aggregation. The ADP (1 µM)-induced phosphorylation of HSP at Ser-78 observed in the platelets from Group 1 was inhibited by PD98059 or SB203580. The use of aspirin ameliorated the accelerated microaggregation of platelets in Group 1, and the low-dose ADP-induced phosphorylation of HSP27 at Ser-78 was no longer observed. These results strongly suggest that the phosphorylation of HSP27 at Ser-78 is correlated with the acceleration of platelet aggregation induced by ADP in type 2 DM patients.

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