Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, Gimhae 621-749.
Department of Biomedical Laboratory Science, College of Medical Science, Konyang University, Daejeon 302-718.
Biomol Ther (Seoul). 2014 May;22(3):223-31. doi: 10.4062/biomolther.2014.025.
In this study, we prepared cordycepin-enriched (CE)-WIB801C, a n-butanol extract of Cordyceps militaris-hypha, and investigated the effect of CE-WIB801C on collagen-induced human platelet aggregation. CE-WIB801C dose-dependently inhibited collagen-induced platelet aggregation, and its IC50 value was 175 μg/ml. CE-WIB801C increased cAMP level more than cGMP level, but inhibited collagen-elevated [Ca(2+)]i mobilization and thromboxane A2 (TXA2) production. cAMP-dependent protein kinase (A-kinase) inhibitor Rp-8-Br-cAMPS increased the CE-WIB801C-downregulated [Ca(2+)]i level in a dose dependent manner, and strongly inhibited CE-WIB801C-induced inositol 1, 4, 5-trisphosphate receptor (IP3R) phosphorylation. These results suggest that the inhibition of [Ca(2+)]i mobilization by CE-WIB801C is resulted from the cAMP/A-kinase-dependent phosphorylation of IP3R. CE-WIB801C suppressed TXA2 production, but did not inhibit the activities of cyclooxygenase-1 (COX-1) and TXA2 synthase (TXAS). These results suggest that the inhibition of TXA2 production by WIB801C is not resulted from the direct inhibition of COX-1 and TXAS. In this study, we demonstrate that CE-WIB801C with cAMP-dependent Ca(2+)-antagonistic antiplatelet effects may have preventive or therapeutic potential for platelet aggregation-mediated diseases, such as thrombosis, myocardial infarction, atherosclerosis, and ischemic cerebrovascular disease.
在这项研究中，我们制备了虫草素富集（CE）-WIB801C，一种蛹虫草菌丝体的正丁醇提取物，并研究了 CE-WIB801C 对胶原诱导的人血小板聚集的影响。CE-WIB801C 呈剂量依赖性抑制胶原诱导的血小板聚集，其 IC50 值为 175μg/ml。CE-WIB801C 增加 cAMP 水平的作用大于 cGMP 水平，但抑制胶原升高的 [Ca(2+)]i 动员和血栓烷 A2（TXA2）的产生。cAMP 依赖性蛋白激酶（A-激酶）抑制剂 Rp-8-Br-cAMPS 呈剂量依赖性增加 CE-WIB801C 下调的 [Ca(2+)]i 水平，并强烈抑制 CE-WIB801C 诱导的肌醇 1,4,5-三磷酸受体（IP3R）磷酸化。这些结果表明，CE-WIB801C 抑制 [Ca(2+)]i 动员是由 cAMP/A-激酶依赖性 IP3R 磷酸化引起的。CE-WIB801C 抑制 TXA2 的产生，但不抑制环加氧酶-1（COX-1）和 TXA2 合酶（TXAS）的活性。这些结果表明，WIB801C 抑制 TXA2 的产生不是由于直接抑制 COX-1 和 TXAS。在这项研究中，我们证明了具有 cAMP 依赖性 Ca(2+)-拮抗抗血小板作用的 CE-WIB801C 可能对血小板聚集介导的疾病，如血栓形成、心肌梗死、动脉粥样硬化和缺血性脑血管疾病具有预防或治疗潜力。
