蛭蛇通络胶囊及其拆方组分肠吸收液抗氧化及抗凝血研究  

Study on Antioxidation and Anticoagulation of Intestinal Absorption Solution of Zhishe Tongluo Capsule(蛭蛇通络胶囊)and Its Disassembled Prescriptions'Components

作　　者：丁茹 李煜 张毅[1] 王丽芳[1] 张方博[1] 杨洪军[4] DING Ru;LI Yu;ZHANG Yi;WANG Li-fang;ZHANG Fang-bo;YANG Hong-jun(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beiing 100700,China;Shanxi Jianmin Pharmaceutical Co.,LTD,Xianyang Shanxi 712000,China;Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院中药研究所,北京100700 [2]陕西健民制药有限公司,咸阳陕西712000 [3]天津中医药大学,天津301617 [4]中国中医科学院医学实验中心,北京100700

出　　处：《中医药导报》2022年第2期1-6,12,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：陕西省重点研发计划项目(2019ZDLSF04-04-01);国家“重大新药创制”科技重大专项(2019ZX09201005)。

摘　　要：目的:通过肠吸收液-体外药理活性结合拆方研究蛭蛇通络胶囊抗氧化和抗凝的主要组分。方法:根据制备工艺路线将蛭蛇通络胶囊进行拆方,得到全方和3个组分,分别制备相应的肠吸收液。通过DPPH自由基清除实验观察抗氧化作用,通过凝血四项和ADP诱导血小板凝集实验观察抗凝作用。构建氧糖剥夺SH-SY5Y细胞模型,验证蛭蛇通络胶囊全方及拆方组分通过抑制氧化应激发挥神经保护作用。结果:蛭蛇通络胶囊能清除DPPH自由基,最强的成分为含人参、黄芪、天麻、丹参和葛根的组分1;降低纤维蛋白原(FIB)水平、延长凝血酶时间(TT)并抑制ADP诱导的血小板凝集最强的成分为含水蛭、乌梢蛇和红花的组分2。蛭蛇通络胶囊全方和含人参、黄芪、天麻、丹参和葛根的组分1均能抑制氧糖剥夺SH-SY5Y细胞内乳酸脱氢酶(LDH)释放,升高超氧化物歧化酶(SOD)活性,降低丙二醛(MDA)水平,抑制神经细胞损伤,提高细胞生存率。结论:通过制备工艺路线将蛭蛇通络胶囊进行快速拆解,寻找并定位全方中发挥抗氧化和抗凝的主要组分。结果证实蛭蛇通络胶囊清除DPPH自由基,发挥抗氧化作用以含人参、黄芪、天麻、丹参和葛根的组分为最佳;抗凝血作用以含水蛭、乌梢蛇和红花的组分最佳。本研究为中药复方的功效物质基础提供了实验证据,但其有效成分和具体机制仍需深入研究。Objective:To study the antioxidant and anticoagulant components of Zhishe Tongluo Capsule(ZSTLC)by combining intestinal absorption solution with pharmacological activity in vitro.Method:The prescription of ZSTLC was separated by the production process,and the corresponding intestinal absorption solutions of three separated and whole prescriptions were obtained.The antioxidant effect was observed by DPPH free radical scavenging test,and the anticoagulant effect was observed by four items of coagulation and ADP induced platelet aggregation test.The oxygen glucose deprivation SH-SY5Y cell model was constructed to verify that the whole formula and disassembled components of ZSTLC play a neuroprotective role by inhibiting oxidative stress.Results:ZSTLC can scavenge DPPH free radicals,and the strongest active ingredient were in component 1,which containing Renshen(Ginseng),Huangqi(Astragalus),Tianma(Gastrodia elata),Danshen(Salvia miltiorrhiza)and Gegen(Pueraria).The strongest active ingredient in component 2 which containing Shuizhi(Leech),Wushaoshe(Zaocys)and Honghua(carthamus),can decreased the level of fibrinogen(FIB),prolonged thrombin time(TT)and inhibited ADP induced platelet aggregation.ZSTLC and component 1 containing Renshen(Ginseng),Huangqi(Astragalus),Tianma(Gastrodia elata),Danshen(Salvia miltiorrhiza)and Gegen(Pueraria)can inhibit the release of lactate dehydrogenase(LDH)in oxygen glucose deprived SH-SY5Y cells,increase the activity of superoxide dismutase(SOD),reduce the level of malondialdehyde(MDA),inhibit nerve cell damage and improve cell survival rate.Conclusion:According to the production process,ZSTLC were quickly disassembled to find the main components of anti-oxidation and anti-coagulation.The results showed that component containing Renshen(Ginseng),Huangqi(Astragalus),Tianma(Gastrodia elata),Danshen(Salvia miltiorrhiza)and Gegen(Pueraria)in ZSTLC was the best one to scavenging DPPH free radical and exerting antioxidant effect;The best anticoagulant effect was the component containing Shuizhi(

关 键 词：蛭蛇通络胶囊 肠吸收液 抗氧化 抗凝血 DPPH自由基 凝血四项 血小板凝集 氧糖剥夺 有效成分辨识 SH-SY5Y细胞 

分 类 号：R285.5[医药卫生—中药学]