1. 湖北医药学院, a. 药学院; b. 武当特色中药研究湖北省重点实验室, 湖北 十堰 442000; 2. 太和医院干细胞研究中心, 湖北 十堰 442000
Establishment of Platelet Membrane Chromatography and Column Temperature Study
LI Ou1a, FU Zhen-zhen1a, LIU Kang-ni1a,1b,2, LEI Yu-tian1a, LI Jia-rui1a, GAO Xiao-kang1a,1b,2*
1a. College of Pharmacy; 1b. The Open Project of Hubei Key Laboratory of Wudang Local Chinese Medicine Research, Hubei University of Medicine, Shiyan 442000, China; 2. Taihe Hospital Stem Cell Research Center, Shiyan 442000, China)
Abstract：OBJECTIVE To establish a platelet cell membrane chromatographic model and investigate the retention behaviors of anti-platelet aggregation drugs on chromatographic column at different temperatures, and simulate the interactions between drugs and platelets under normal and febrile pathological conditions. METHODS The platelet cell membrane chromatographic stationary phase was constructed by physical adsorption method. The column was packed with wet method. The protein content was determined by BCA protein concentration assay kit. The biological activity was determined by Na+, K+-ATPase assay kit. The chromatographic model was used to investigate the specificity of the column and the retention characteristics of drugs in the temperature range of 35.0-42.0 ℃. RESULTS The activity of platelet ATPase was 0.214, and the concentration of platelet membrane protein was 0.340 9 mg·mL-1 before bonding and 0.080 5 mg·mL-1 after bonding. The retention characteristics of clopidogrel, dipyridamole and cilostazole on platelet cell membrane chromatographic column and blank silica gel column were quite different. The retention time of the three drugs on platelet cell membrane chromatographic column was the maximum at 36.0 ℃, and then decreased with the increase of temperature. CONCLUSION A platelet cell membrane chromatographic model is successfully constructed, and the retention characteristics of antiplatelet aggregates at different temperatures are studied for the first time. The chromatographic retention behaviors of antiplatelet aggregates at normal and febrile body temperatures are simulated.
李欧, 付真真, 刘康妮, 雷雨甜, 李佳瑞, 高小康. 血小板细胞膜色谱构建及柱温研究[J]. 中国药学杂志, 2020, 55(15): 1289-1294.
LI Ou, FU Zhen-zhen, LIU Kang-ni, LEI Yu-tian, LI Jia-rui, GAO Xiao-kang. Establishment of Platelet Membrane Chromatography and Column Temperature Study. Chinese Pharmaceutical Journal, 2020, 55(15): 1289-1294.
LU X H, JIN N Z. Application of biochromatography inscreening the biologically active principles of Chinese traditional drug . Chin J Biochem Pharm(中国生化药物杂志), 2003,24(5):1-23.
[2]
YANG X X, GU W, LIANG L, et al. Screening for the bioactive constituents of traditional Chinese medicines-progress and challenges. RSC Adv, 2017, 7:3089-3100.
[3]
LI C Q, HE L C, DONG H Y, et al. Screening for the anti-inflammatory activity of fractions and compounds from Atractylodes macrocephala koidz. J Ethnopharmacol,2007, 114 (2):212-217.
[4]
LI Y P, HE L C. Establishment of the model of vascular endothelial cell membrane chromatography and its preliminary application . Chin Sci Bull(科学通报),2007,52(7):922-928.
[5]
XU X Y, WANG S M, CHEN W H, et al. Effects of Taohong Siwu decoction II in the chick chorioallantoic membrane assay and on B16 melanoma in mice and endothelial cells ECV304 proliferation . J Tradit Chin Med(中医杂志), 2006, 26(1):63-67.
[6]
WANG Q, XING M, CHEN W H, et al. HPLC-APCI-MS/MS method for the determination of catalpol in rat plasma and cerebrospinal fluid:application to an in vivo pharmacokinetic study . J Pharm Biomed Anal, 2012, 11(70):337-343.
[7]
LI S P, ZHAO J, QIAN Z M, et al. Advanced development of chromatography in screening and identification of effective compounds in Chinese material medica . Sci Sin:Chem, 2010, 40(6):651-667.
[8]
QIU H X, HUANG Q D, CHEN D, et al. Application progress on the biochromatography in traditional Chinese medicine research . Chin Arch Tradit Chin Med (中华中医药学刊), 2010, 28(1):144-147.
[9]
SHI X L, FU J, LI G W. Advances in studies on active components and monomers of platelet-targeted Chinese medicines . China J China Mater Med(中国中药杂志), 2006, 31(5):361-365. LIANG R F, XU L, YUE M Q. Advances in the application of cell fragmentation technology . Inner Mongolia Agricul Sci Technol(内蒙古农业科技), 2013, 2(1):113-114. YANG Y F, ZHAO D D. Methods and applications of cell membrane fragmentation technology . Rural Economy Technol(农村经济与科技), 2017,28(12):300. MAGDALENA K O, MAGORZATA B R, EWA Z D, et al. Comparative study of fungal cell disruption-scope and limitations of the methods. Folia Microbiol, 2011, 56 (5):469-475. LIANG X J, WANG S, LIU S J, et al. A novel octadecylsilane functionalized graphene oxide/silica composite stationary phase for high performance liquid chromatography . J Separation Sci, 2012, 35(16):2003-2009. PRAJITHA N, ATHIRA S S, MOHANANP V, et al. A polypeptide produces fever by metabolic changes in hypothalamus . Immunol Lett, 2018, 12(204):38-46. BLOMQVIST A, ENGBLOM D. Neural mechanisms of inflammation induced fever . Neuroscientist, 2018, 24(4):381-399. XING J H, YANG L Y, LI Q, et al. Advances in antithrombotic drugs . Prog Pharm Sci (药学进展), 2014, 38(3):174-184. ZHANG J C,ZI Y. Clinical progress of antiplatelet aggregation drugs in the treatment of ischemic stroke . Pract J Clin Med(实用临床医药杂志), 2012, 16(17):175-178.
2020, Vol. 55 
