title: Antithrombosis activity of protocatechuic and shikimic acids from functional plant Pinus densiflora Sieb. et Zucc needles.
abstract: Pine needle extract (PE) and fermented pine needle extract (FPE) have been reported to show various biological and pharmacological activities such as antioxidant, anti-inflammatory, anti-bacterial, anti-cholesterol, gastrointestinal motility control, and fibrinolytic effect. The aims of our research were to isolate fibrinolytic compounds from PE and FPE and evaluate their antithrombotic activity in vitro and in vivo. Protocatechuic (1) and shikimic (2) acids were isolated and identified from FPE. 1 and 2 not only have fibrinolysis activity but also inhibit fibrin formation similar to aspirin. Lysis of fibrin clots by 1 and 2 occurred completely at pH 2-4. Results of SDS-PAGE showed that fibrin polypeptide chains (Aα, Bβ, γ) lysed by 1 and 2 were intact. The antithrombotic effects of 1 and 2 were confirmed by models of carrageenan-induced tail thrombosis, collagen and epinephrine-induced pulmonary thromboembolism in mice, and FeCl3-induced carotid arterial thrombus. Moreover, 1 and 2 did not induce hemorrhage in the tail veins of mice, unlike common antithrombotic compounds. We also measured changes in the quantities of 1 and 2 obtained from FPE. As fermentation progressed, we demonstrated that the quantity of 1 steadily increased, while the quantity of 2 did not significantly change. We therefore demonstrated that FPE is an excellent resource for 1 and 2 and can be produced inexpensively in sufficient quantities for industrial-scale extraction.
authors.affiliation: Department of Biomedical Science and BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju, 501-759, Korea.
authors.forename: Jaeyoung
authors.initials: J
authors.lastname: Park
authors.affiliation: Department of Biomedical Science and BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju, 501-759, Korea.
authors.forename: Beomgi
authors.initials: B
authors.lastname: Lee
authors.affiliation: Department of Biomedical Science and BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju, 501-759, Korea.
authors.forename: Hakjoon
authors.initials: H
authors.lastname: Choi
authors.affiliation: Department of Biomedical Science and BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju, 501-759, Korea.
authors.forename: Woong
authors.initials: W
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Chosun University, Gwangju, 501-759, Korea.
authors.forename: Ho-Joong
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Biomedical Science and BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju, 501-759, Korea. hscheong@chosun.ac.kr.
authors.forename: Hyeonsook
authors.initials: H
authors.lastname: Cheong
mesh_terms: D000818:Animals; D005343:Fibrinolytic Agents / Q000627:therapeutic use*; D062385:Hydroxybenzoates / Q000737:chemistry*; D008297:Male; D051379:Mice; D028223:Pinus; D010936:Plant Extracts / Q000737:chemistry*; D051381:Rats; D012765:Shikimic Acid / Q000737:chemistry*; D013927:Thrombosis / Q000188:drug therapy*
chemical_list: D005343:Fibrinolytic Agents; D062385:Hydroxybenzoates; D010936:Plant Extracts; D012765:Shikimic Acid; C009091:protocatechuic acid
keywords: Fermented pine needle extract; Fibrinolysis; Pinus densiflora Sieb. et Zucc; Protocatechuic acid; Shikimic acid