title: Antiplatelet and antithrombotic effects of cordycepin-enriched WIB-801CE from Cordyceps militaris ex vivo, in vivo, and in vitro.
abstract: BACKGROUND
  A species of the fungal genus Cordyceps has been used as a complementary and alternative medicine of traditional Chinese medicine, and its major component cordycepin and cordycepin-enriched WIB-801CE are known to have antiplatelet effects in vitro. However, it is unknown whether they have also endogenous antiplatelet and antithrombotic effects. In this study, to resolve these doubts, we prepared cordycepin-enriched WIB-801CE, an ethanol extract from Cordyceps militaris-hypha, then evaluated its ex vivo, in vivo, and in vitro antiplatelet and antithrombotic effects.
  
  
  METHODS
  Ex vivo effects of WIB-801CE on collagen- and ADP-induced platelet aggregation, serotonin release, thromboxane A2(TXA2) production and its associated activities of enzymes [cyclooxygenase-1 (COX-1), TXA2synthase (TXAS)], arachidonic acid (AA) release and its associated phosphorylation of phospholipase Cβ3, phospholipase Cγ2or cytosolic phospholipase A2, mitogen-activated protein kinase (MAPK) [p38 MAPK, extracellular signal-regulated kinase (ERK)], and blood coagulation time in rats were investigated. In vivo effects of WIB-801CE on collagen plus epinephrine-induced acute pulmonary thromboembolism, and tail bleeding time in mice were also inquired. In vitro effects of WIB-801CE on cytotoxicity, and fibrin clot retraction in human platelets, and nitric oxide (NO) production in RAW264.7 cells or free radical scavenging activity were studied.
  
  
  RESULTS
  Cordycepin-enriched WIB-801CE inhibited ex vivo platelet aggregation, TXA2production, AA release, TXAS activity, serotonin release, and p38 MAPKand ERK2 phosphorylation in collagen- and ADP-activated rat platelets without affecting blood coagulation. Furthermore, WIB-801CE manifested in vivo inhibitory effect on collagen plus epinephrine-induced pulmonary thromboembolism mice model. WIB-801CE inhibited in vitro NO production and fibrin clot retraction, but elevated free radical scavenging activity without affecting cytotoxicity against human platelets.
  
  
  CONCLUSIONS
  WIB-801CE inhibited collagen- and ADP-induced platelet activation and its associated thrombus formation ex vivo and in vivo. These were resulted from down-regulation of TXA2production and its related AA release and TXAS activity, and p38MAPKand ERK2 activation. These results suggest that WIB-801CE has therapeutic potential to treat platelet activation-mediated thrombotic diseases in vivo.
authors.affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, 197, Inje-ro, Gyungnam, Gimhae, 50834, Korea.
authors.forename: Hyuk-Woo
authors.initials: HW
authors.lastname: Kwon
authors.affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, 197, Inje-ro, Gyungnam, Gimhae, 50834, Korea.
authors.forename: Jung-Hae
authors.initials: JH
authors.lastname: Shin
authors.affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, 197, Inje-ro, Gyungnam, Gimhae, 50834, Korea.
authors.forename: Deok Hwi
authors.initials: DH
authors.lastname: Lim
authors.affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, 197, Inje-ro, Gyungnam, Gimhae, 50834, Korea.
authors.forename: Woo Jeong
authors.initials: WJ
authors.lastname: Ok
authors.affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, 197, Inje-ro, Gyungnam, Gimhae, 50834, Korea.
authors.forename: Gi Suk
authors.initials: GS
authors.lastname: Nam
authors.affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, 197, Inje-ro, Gyungnam, Gimhae, 50834, Korea.
authors.forename: Min Ji
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: Central Research Center, Whanin Pharm. Co., Ltd., 107, Gwanggyo-ro, Suwon, Gyeonggi-do, 16229, Korea.
authors.forename: Ho-Kyun
authors.initials: HK
authors.lastname: Kwon
authors.affiliation: Central Research Center, Whanin Pharm. Co., Ltd., 107, Gwanggyo-ro, Suwon, Gyeonggi-do, 16229, Korea.
authors.forename: Jun-Hee
authors.initials: JH
authors.lastname: Noh
authors.affiliation: Central Research Center, Whanin Pharm. Co., Ltd., 107, Gwanggyo-ro, Suwon, Gyeonggi-do, 16229, Korea.
authors.forename: Je-Young
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, 197, Inje-ro, Gyungnam, Gimhae, 50834, Korea.
authors.forename: Hyun-Hong
authors.initials: HH
authors.lastname: Kim
authors.affiliation: Central Research Center, Whanin Pharm. Co., Ltd., 107, Gwanggyo-ro, Suwon, Gyeonggi-do, 16229, Korea. kimjl@whanin.com.
authors.forename: Jong-Lae
authors.initials: JL
authors.lastname: Kim
authors.affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, 197, Inje-ro, Gyungnam, Gimhae, 50834, Korea. mlsjpark@inje.ac.kr.
authors.forename: Hwa-Jin
authors.initials: HJ
authors.lastname: Park
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001777:Blood Coagulation / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D032661:Cordyceps / Q000737:chemistry*; D004353:Drug Evaluation, Preclinical; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D008297:Male; D008813:Mice, Inbred ICR; D009569:Nitric Oxide / Q000378:metabolism; D010766:Phosphorylation; D010936:Plant Extracts / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D017207:Rats, Sprague-Dawley; D012701:Serotonin / Q000378:metabolism; D013928:Thromboxane A2 / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D005343:Fibrinolytic Agents; D010936:Plant Extracts; D010975:Platelet Aggregation Inhibitors; C000605556:WIB801C; D016718:Arachidonic Acid; D009569:Nitric Oxide; D012701:Serotonin; D013928:Thromboxane A2; D010738:Type C Phospholipases; D002118:Calcium
keywords: Arachidonic acid release; Cordycepin; ERK2; Platelet aggregation; Serotonin; TXA2; Thromboxane A2 synthase; Thrombus; WIB-801CE; p38 MAPK