丹红化瘀口服液对视网膜中央静脉阻塞症关键靶点的调节作用  

Regulatory effect of Danhong Huayu Oral Liquid on key targets of central retinal vein occlusion

作　　者：韩彦琪[1,2] 武琦 林娟[4] 刘耀晨 许浚[1,2,6] 张洪兵[1,2] 张铁军[1,2] 刘昌孝[1,2,6] HAN Yan-qi;WU Qi;LIN Juan;LIU Yao-chen;XU Jun;ZHANG Hong-bing;ZHANG Tie-jun;LIU Chang-xiao(Tianjin Key Laboratory of Quality Marker of Traditional Chinese Medicine,Tianjin Institute of Pharmaceutical Research,Tianjin 300462,China;State Key Laboratory of Drug Delivery Technology and Pharmacokinetics,Tianjin Institute of Pharmaceutical Research,Tianjin 300462,China;Pharmaron Beijing Co.,Ltd.,Beijing 100176,China;Guangzhou Baiyunshan Hutchison Whampoa Chinese Medicine Co.,Ltd.,Guangzhou 510515,China;Tianjin Medical University,Tianjin 300070,China;Research Unit for Drug Metabolism,Chinese Academy of Medical Sciences,Tianjin Institute of Pharmaceutical Research,Tianjin 300462,China)

机构地区：[1]天津药物研究院天津市中药质量标志物重点实验室,天津300462 [2]天津药物研究院释药技术与药物代谢动力学国家重点实验室,天津300462 [3]康龙化成(北京)生物技术有限公司,北京100176 [4]广州白云山和记黄埔中药有限公司,广东广州510515 [5]天津医科大学,天津300070 [6]天津药物研究院中国医学科学院药物代谢新技术创新单元,天津300462

出　　处：《中草药》2022年第6期1634-1642,共9页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81830111);中医药现代化研究重点专项(2019YFC1711200);天津市自然科学基金资助项目(19JCQNJC12700);广西中药药效研究重点实验室开放课题(20-065-38);中国医学科学院医学与健康科技创新工程项目资助(2019-I2M-5-020)。

摘　　要：目的采用G蛋白偶联受体及酶活检测实验方法,探究丹红化瘀口服液对视网膜中央静脉阻塞症关键靶点的调节作用,明确其作用机制。方法选取磷酸二酯酶3A(phosphodiesterase 3A,PDE3A)、凝血酶(Thrombin)、内皮素转化酶1(endothelin converting enzyme 1,ECE1)、血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2)、α1A肾上腺素受体(α1A adrenergic receptor,ADRA1A)为研究载体,通过胞内钙离子荧光检测和酶抑制剂检测技术评价丹红化瘀口服液及14个主要单体成分对PDE3A、Thrombin、ECE1、VEGFR2酪氨酸激酶和ADRA1A的抑制活性。结果丹红化瘀口服液对以上5个靶点均有显著的抑制作用(P<0.001);丹酚酸D、丹酚酸B、丹参素、迷迭香酸和柚皮苷对PDE3A有显著的抑制作用(P<0.05、0.001);14个化合物对Thrombin均有显著的抑制活性(P<0.05、0.01、0.001);丹酚酸D、丹酚酸B、丹参素、迷迭香酸、阿魏酸、羟基红花黄色素A对ECE1有显著的抑制活性(P<0.05、0.01、0.001);丹酚酸D、丹酚酸B、丹参素、迷迭香酸、柠檬苦素对VEGFR2酪氨酸激酶有显著的抑制效果(P<0.01、0.001);柴胡皂苷a可显著抑制ADRA1A活性(P<0.001)。结论 ADRA1A、VEGFR2、PDE3A、Thrombin和ECE1可能为丹红化瘀口服液治疗中央静脉阻塞症的关键作用靶点,其药效物质基础可能为丹参酮IIA、丹酚酸B、丹酚酸D、丹参素、迷迭香酸、原儿茶醛、阿魏酸、川芎嗪、洋川芎内酯I、羟基红花黄色素A、苦杏仁苷、柴胡皂苷a、柚皮苷和柠檬苦素。Objective To explore the regulatory effect and mechanism of Danhong Huayu Oral Liquid(丹红化瘀口服液,DHOL)on key targets of central retinal vein occlusion(CRVO)by G protein coupled receptor(GPCR)and enzyme activity assay.Methods Phosphodiesterase 3 A(PDE3 A),thrombin,endothelin converting enzyme 1(ECE1),vascular endothelial growth factor receptor 2(VEGFR2)andα1 A adrenergic receptor(ADRA1 A)were selected as research vectors.Intracellular calcium fluorescence detection and enzyme inhibitor detection techniques were used to evaluate the inhibitory effects of DHOL and 14 main compounds on ADRA1 A,VEGFR2 tyrosine kinase,PDE3 A,thrombin and ECE1.Results DHOL significantly inhibited the above five targets(P<0.001);Salvianolic acid D,salvianolic acid B,danshensu,rosmarinic acid and naringin significantly inhibited PDE3 A(P<0.05,0.001);All the 14 compounds had significant inhibitory effects on thrombin(P<0.05,0.01,0.001);Salvianolic acid D,salvianolic acid B,danshensu,rosmarinic acid,ferulic acid and hydroxysafflor yellow A had inhibitory activities on ECE1(P<0.05,0.01,0.001);Salvianolic acid D,salvianolic acid B,danshensu,rosmarinic acid and limonin had inhibitory effects on VEGFR2 tyrosine kinase(P<0.01,0.001);Saikosaponin a had significant antagonize on ADRA1 A(P<0.001).Conclusion ADRA1 A,VEGFR2,PDE3 A,thrombin and ECE1 may be the key targets of DHOL in the treatment of CRVO.The pharmacodynamic material basis may be tanshinone IIA,salvianolic acid B,salvianolic acid D,danshensu,rosmarinic acid,protocatechualdehyde,ferulic acid,ligustrazine,senkyunolideⅠ,hydroxysafflor yellow A,amygdalin,saikosaponin a,naringin and limonin.

关 键 词：丹红化瘀口服液 视网膜中央静脉阻塞症 磷酸二酯酶 凝血酶 内皮素转化酶 血管内皮生长因子受体2 Α1肾上腺素受体 丹参酮IIA 丹酚酸B 丹酚酸D 丹参素 迷迭香酸 原儿茶醛 阿魏酸 川芎嗪 洋川芎内酯Ⅰ 羟基红花黄色素A 苦杏仁苷 柴胡皂苷A 柚皮苷 柠檬苦素 

分 类 号：R285.5[医药卫生—中药学]