title: Release of Phosphorylated HSP27 (HSPB1) from Platelets Is Accompanied with the Acceleration of Aggregation in Diabetic Patients.
abstract: We investigated the relationship between HSP27 phosphorylation and collagen-stimulated activation of platelets in patients with diabetes mellitus (DM). Platelet-rich plasma was prepared from blood of type 2 DM patients. The platelet aggregation was analyzed in size of aggregates by an aggregometer using a laser scattering method. The protein phosphorylation was analyzed by Western blotting. Phosphorylated-HSP27 and PDGF-AB released from platelets were measured by ELISA. The phosphorylated-HSP27 levels at Ser-78 and Ser-82 induced by collagen were directly proportional to the platelet aggregation. Total HSP27 levels in platelets were decreased concomitantly with the phosphorylation. The released HSP27 levels were significantly correlated with the phosphorylated levels of HSP27 in the platelets stimulated by 0.3 μg/ml collagen. The low dose collagen-stimulated release of HSP27 was detected but relatively small in healthy donors. The released levels of PDGF-AB were in parallel with the levels of released HSP27. Area under the curve (AUC) of small aggregation (9-25 μm) induced by 0.3 μg/ml collagen was inversely proportional to the levels of released HSP27. AUC of large aggregation (50-70 μm) was directly proportional to the levels of released HSP27. Exogenous recombinant phosphorylated- HSP27 hardly affected the aggregation or the released levels of PDGF-AB induced by collagen. These results strongly suggest that HSP27 is released from human platelets accompanied with its phosphorylation induced by collagen, which is correlated with the acceleration of platelet aggregation in type 2 DM patients.
authors.affiliation: Department of Clinical Laboratory, National Center for Geriatrics and Gerontology, Obu, Aichi, Japan; Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Tokuda
authors.affiliation: Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Japan; Department of Orthopedic Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
authors.forename: Gen
authors.initials: G
authors.lastname: Kuroyanagi
authors.affiliation: Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Japan; Department of Neurosurgery, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Masanori
authors.initials: M
authors.lastname: Tsujimoto
authors.affiliation: Department of Neurosurgery, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Enomoto
authors.affiliation: Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Matsushima-Nishiwaki
authors.affiliation: Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Japan; Department of Anesthesiology and Pain Medicine, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Onuma
authors.affiliation: Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Japan; Department of Anesthesiology and Pain Medicine, Gifu University Graduate School of Medicine, Gifu, Japan; Department of Anesthesiology and Critical Care Medicine, Matsunami General Hospital, Gifu, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Kojima
authors.affiliation: Department of Emergency and Disaster Medicine, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Doi
authors.affiliation: Department of Anesthesiology and Pain Medicine, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Tanabe
authors.affiliation: Department of Anesthesiology and Critical Care Medicine, Matsunami General Hospital, Gifu, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Akamatsu
authors.affiliation: Department of Anesthesiology and Pain Medicine, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Iida
authors.affiliation: Department of Emergency and Disaster Medicine, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Ogura
authors.affiliation: Department of Orthopedic Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
authors.forename: Takanobu
authors.initials: T
authors.lastname: Otsuka
authors.affiliation: Department of Neurosurgery, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Iwama
authors.affiliation: Department of Clinical Laboratory, National Center for Geriatrics and Gerontology, Obu, Aichi, Japan.
authors.forename: Takahisa
authors.initials: T
authors.lastname: Tanikawa
authors.affiliation: Department of Clinical Laboratory, National Center for Geriatrics and Gerontology, Obu, Aichi, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Ishikawa
authors.affiliation: Department of Clinical Laboratory, National Center for Geriatrics and Gerontology, Obu, Aichi, Japan.
authors.forename: Kumi
authors.initials: K
authors.lastname: Kojima
authors.affiliation: Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Kozawa
mesh_terms: D000368:Aged; D019540:Area Under Curve; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D003094:Collagen / Q000494:pharmacology*; D003924:Diabetes Mellitus, Type 2 / Q000097:blood / Q000473:pathology; D005260:Female; D055551:HSP27 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006360:Heat-Shock Proteins; D006801:Humans; D008297:Male; D008875:Middle Aged; D018832:Molecular Chaperones; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects*; D010982:Platelet-Derived Growth Factor / Q000235:genetics / Q000378:metabolism; D053657:Platelet-Rich Plasma / Q000737:chemistry; D061251:Primary Cell Culture; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction
chemical_list: D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D010982:Platelet-Derived Growth Factor; C095477:platelet-derived growth factor AB; D003094:Collagen