血小板细胞膜固相色谱法的建立及其对丹参效应物质的初步分析

Establishment of platelet membrane immobilized chromatography and its application in the analysis of active components of Radix Salviae Miltiorrhizae

目的 建立血小板细胞膜固相色谱法,并应用其分析丹参对血小板作用的活性成分。方法 应用血小板细胞膜作为固定相选择性地结合中药提取液中的活性成分,洗去未结合的其他成分后,使细胞膜靶点失活,被结合的活性成分从血小板细胞膜中释放出来,样品经固相萃取后用HPLC对这些成分进行解析。结果 丹参水提液的效应成分初步确定为6个,其中之一为原儿茶醛。结论 血小板细胞膜固相色谱法基本可以反映化合物与细胞膜生物靶点(受体、通道、酶等)的相互作用;该体系中的保留成分和其药理作用有显著的相关性。

OBJECTIVE: To establish the platelet membrane immobilized chromatograghy (PMBC) for the effective chemicals analysis of tradition Chinese medicine on platelets. METHODS: The active chemicals of Radix Salviae Mittiorrhizzae were combined with the platelet membrane under imitated physical environments. The unattached substances were washed away with PBS of pH 7.4. Then the attached compounds were eluted by PBS of pH 4.0. These compounds released from target sites were collected and handled through SPE to be condensed. The concentrated solution was analyzed by HPLC. RESULTS: The results indicated that six characteristic active compounds in the water extracts of Radix Salviae, Miltiorrhizae, binded to the membranes of platelets. One of them was protocatechuicaldehyde. The others were water dissolved salvianolic acids. CONCLUSION: PMBC can binding the effective chemicals through platelet membranes (such as receptors, channels, enzymes etc.) according to chromatographic analysis. There was a significant correlation between the retention properties of the active compounds which was obtained by PMBC and their pharmacological action on platelets.