Park Jaeyoung, Lee Beomgi, Choi Hakjoon, Kim Woong, Kim Ho-Joong, Cheong Hyeonsook
Department of Biomedical Science and BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju, 501-759, Korea.
Department of Chemistry, Chosun University, Gwangju, 501-759, Korea.
J Nat Med. 2016 Jul;70(3):492-501. doi: 10.1007/s11418-015-0956-y. Epub 2016 Mar 3.
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.
据报道，松针提取物（PE）和发酵松针提取物（FPE）具有多种生物学和药理活性，如抗氧化、抗炎、抗菌、抗胆固醇、控制胃肠蠕动以及纤溶作用。我们研究的目的是从PE和FPE中分离出纤溶化合物，并评估它们在体外和体内的抗血栓活性。从FPE中分离并鉴定出了原儿茶酸（1）和莽草酸（2）。1和2不仅具有纤溶活性，而且与阿司匹林类似，还能抑制纤维蛋白形成。1和2在pH 2 - 4时能完全溶解纤维蛋白凝块。SDS - PAGE结果表明，1和2溶解的纤维蛋白多肽链（Aα、Bβ、γ）是完整的。通过角叉菜胶诱导的小鼠尾部血栓形成模型、胶原和肾上腺素诱导的小鼠肺血栓栓塞模型以及FeCl3诱导的小鼠颈动脉血栓模型，证实了1和2的抗血栓作用。此外，与常见的抗血栓化合物不同，1和2不会在小鼠尾静脉中引起出血。我们还测量了从FPE中获得的1和2的含量变化。随着发酵的进行，我们证明1的含量稳步增加，而2的含量没有显著变化。因此，我们证明FPE是1和2的优质来源，并且可以以低成本大量生产用于工业规模提取。
