脑心通胶囊肠吸收液抗ADP诱导血小板凝集的有效成分辨识研究
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摘要
应用外翻肠囊法制备肠吸收液,同时拆方或构建重组方,生化试剂盒结合酶标仪检测ADP诱导血小板的聚集率,寻找脑心通胶囊组方中拮抗血小板聚集的单味或多味中药。研究表明脑心通胶囊能抑制ADP诱导血小板聚集。通过拆方以及重组方研究,表明丹参、赤芍、桂枝以及水蛭是脑心通全方抗血小板聚集的主要活性中药。此4味中药又以桂枝在全方中发挥主要的抗血小板聚集作用。桂枝肠吸收液来源的香豆素在50～200μmol·L~(-1)时拮抗血小板聚集。综上所述,丹参、赤芍、桂枝以及水蛭是脑心通胶囊抗ADP诱导血小板聚集的主要活性成分,以桂枝抗血小板聚集作用最强。
Intestinal absorption liquid was prepared by using everted intestinal sac method; meanwhile, its recipes were decomposed or restructured. Platelet aggregation activity was examined by biochemical tests and a microplate reader. One or more kinds of Chinese medicines which displayed inhibiting activity in Naoxintong Capsules were screened through separation and combination of prescription. The results showed that Naoxintong Capsules could inhibit ADP-induced platelet aggregation. Recipe decomposition and restructuring results showed that Salviae Miltiorrhizae Radix et Rhizoma, Paeoniae Radix Rubra, Cinnamomi Ramulus and Hirudo were the main effective medicines in inhibiting platelet aggregation. Furthermore, Cinnamomi Ramulus played a vital role in inhibiting activity among those four kinds of Chinese medicines. Coumarin derived from intestinal absorption liquid of Cinnamomi Ramulus had inhibiting activity in the range of 50-200 μmol·L~(-1), and other ingredients such as cinnamyl alcohol and cinnamaldehyde also had inhibiting activities. In conclusion, Salviae Miltiorrhizae Radix et Rhizoma, Paeoniae Radix Rubra, Cinnamomi Ramulus and Hirudo are the main components for inhibiting ADP-induced platelet aggregation, and Cinnamomi Ramulus has the most strongest inhibiting activity in Naoxintong Capsules.
Intestinal absorption liquid was prepared by using everted intestinal sac method; meanwhile, its recipes were decomposed or restructured. Platelet aggregation activity was examined by biochemical tests and a microplate reader. One or more kinds of Chinese medicines which displayed inhibiting activity in Naoxintong Capsules were screened through separation and combination of prescription. The results showed that Naoxintong Capsules could inhibit ADP-induced platelet aggregation. Recipe decomposition and restructuring results showed that Salviae Miltiorrhizae Radix et Rhizoma, Paeoniae Radix Rubra, Cinnamomi Ramulus and Hirudo were the main effective medicines in inhibiting platelet aggregation. Furthermore, Cinnamomi Ramulus played a vital role in inhibiting activity among those four kinds of Chinese medicines. Coumarin derived from intestinal absorption liquid of Cinnamomi Ramulus had inhibiting activity in the range of 50-200 μmol·L~(-1), and other ingredients such as cinnamyl alcohol and cinnamaldehyde also had inhibiting activities. In conclusion, Salviae Miltiorrhizae Radix et Rhizoma, Paeoniae Radix Rubra, Cinnamomi Ramulus and Hirudo are the main components for inhibiting ADP-induced platelet aggregation, and Cinnamomi Ramulus has the most strongest inhibiting activity in Naoxintong Capsules.
引文
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[11] 郭冬, 刘珍, 张恩户. 丹参酮ⅡA、丹参素、川芎嗪和阿魏酸抑制血小板聚集的相互作用类型及配比关系研究 [J]. 陕西中医, 2007, 28(6): 743.
[12] 邹正午, 徐理纳, 田金英. 迷迭香酸抗血栓和抗血小板聚集作用 [J]. 药学学报, 1993, 25 (4): 241.
[13] 李伟, 张军平, 徐士欣, 等. 丹酚酸 B 对血小板聚集及释放 sP-sel和 sCD40L 的影响 [J]. 中华中医药学刊, 2013, 31(1): 48.
[14] 王琳琳, 丁安伟. 赤芍总苷对大鼠血瘀证模型的影响[J]. 南京中医学院学报, 2011, 27(6): 552.
[15] 黄敬群, 罗晓星, 王四旺, 等. 桂皮醛对抗血小板聚集和血栓形成的特点 [J]. 中国临床康复, 2006, 10(31): 34.
[16] 梁进权, 宓穗卿, 王宁生. 水蛭、虻虫配伍的抗凝血和抗血小板聚集的作用 [J]. 中药材, 2009, 32(9): 1347.
[17] 佘甘树,林育愉,吴钉红.中药抗凝血作用的网络药理学[J].中国实验方剂学杂志,2014,20(16):224.
[2] Choi J H, Cha J K, Huh J T. Adenosine diphosphate-induced platelet aggregation might contribute to poor outcomes in atrial fibrillation-related ischemic stroke [J]. J Stroke Cerebrovasc Dis, 2014, 23(3): e215.
[3] 赵涛, 赵步长, 伍海勤, 等. 脑心通胶囊在心脑血管病中的作用研究 [J]. 中医杂志, 2012, 53(24): 2150.
[4] 范莉, 濮润, 赵海誉, 等. 红花抗 ADP 诱导的血小板聚集活性研究 [J]. 中国中药杂志, 2011, 36(9): 1242.
[5] Wilson T H,Wiseman G.The use of SaCS of evened small intestine for the study of the transference of substances from the mucosal to the serosal surface [J]. Physiol,1954, 123(1): 116.
[6] Patel J P, Korashy H M, El-Kadi A O, et al. Effect of bileand lipids on the stereoselective metabolism of halofantrine by rat everted-intestinal sacs [J]. Chirality, 2010, 22(2): 275.
[7] 黄斌, 陈晓萌, 张迎春, 等. 元胡止痛方肠吸收液对大鼠离体胸主动脉环张力的影响[J]. 中国实验方剂学杂志, 2012, 18(5): 117.
[8] Zhang Y C, Xu H Y, Chen X M, et al. Study on the application of intestinal absorption in vitro coupled with bioactivity assessment in Yuanhu Zhitong preparation [J]. J Med Plants Res, 2012, 6 (10): 1941.
[9] Zhang F B, Huang B, Zhao Y, et al. BNC protects H9c2 cardiomyoblasts from H2O2-induced oxidative injury through ERK1/2 signaling pathway [J]. Evid Based Complement Alternat Med, 2013, doi:10.1155/2013/802784.
[10] 黄斌, 李耿, 郭宇飞, 等.脑心通胶囊中 4 个成分肠吸收研究[J]. 中国中药杂志, 2013, 38(6): 889.
[11] 郭冬, 刘珍, 张恩户. 丹参酮ⅡA、丹参素、川芎嗪和阿魏酸抑制血小板聚集的相互作用类型及配比关系研究 [J]. 陕西中医, 2007, 28(6): 743.
[12] 邹正午, 徐理纳, 田金英. 迷迭香酸抗血栓和抗血小板聚集作用 [J]. 药学学报, 1993, 25 (4): 241.
[13] 李伟, 张军平, 徐士欣, 等. 丹酚酸 B 对血小板聚集及释放 sP-sel和 sCD40L 的影响 [J]. 中华中医药学刊, 2013, 31(1): 48.
[14] 王琳琳, 丁安伟. 赤芍总苷对大鼠血瘀证模型的影响[J]. 南京中医学院学报, 2011, 27(6): 552.
[15] 黄敬群, 罗晓星, 王四旺, 等. 桂皮醛对抗血小板聚集和血栓形成的特点 [J]. 中国临床康复, 2006, 10(31): 34.
[16] 梁进权, 宓穗卿, 王宁生. 水蛭、虻虫配伍的抗凝血和抗血小板聚集的作用 [J]. 中药材, 2009, 32(9): 1347.
[17] 佘甘树,林育愉,吴钉红.中药抗凝血作用的网络药理学[J].中国实验方剂学杂志,2014,20(16):224.