目的采用45Ca跨膜内流测量技术确认复方丹参缓释片及其组分对电压依赖性钙通道(potentitaldependent Ca2+channel,PDC)钙离子内流的拮抗作用,探讨其体外缓释作用的综合评价方法将缓释制剂释放度测定法与45Ca跨膜内流测量技术相结合,在规定的时间点取复方丹参缓释片体外释放样品测定.取Wistar大鼠胸主动脉,分成小段依次置于复合电解质溶液(PSS)、"45Ca-PSS+药"混合液、"45Ca-PSS高钾+药"混合液中,取出动脉环用冷EGTA洗涤,消化.用液体闪烁计数器测量复方丹参缓释片、其组分及体外释放样品的放射性活度.结果复方丹参缓释片、复方丹参片、丹参脂溶性部位、丹参水溶性部位、三七总皂苷、丹参酮ⅡA、丹参素、原儿茶醛、人参皂苷Rb1和三七皂苷R1均有极显著的钙拮抗作用,冰片有显著的钙拮抗作用;复方丹参缓释片4h开始有显著钙拮抗作用,6h后有极显著钙拮抗作用,而自制复方丹参片1h即有显著钙拮抗作用,2h后有极显著钙拮抗作用.结论复方丹参缓释片为钙拮抗剂;丹参脂溶性部位、丹参水溶性部位、三七总皂苷为钙拮抗有效部位,丹参酮ⅡA、丹参素、原儿茶醛、人参皂苷Rb1、三七皂苷R1和冰片为钙拮抗有效成分;与自制复方丹参片比较,复方丹参缓释片具有延缓起效作用,通过体外释放样品钙拮抗作用测定,可以对治疗心血管疾病的中药及天然药物制剂的体外缓释作用进行综合评价.

Objective To study the calcium antagonizing effect of Compound Danshen Sustained— release Tablet(CDST)and its ingredients on potential—dependent calcium channel(PDC)，comprehensive evaluation method of sustained—release effect in vitro by 45Ca determination of transmembrane influx technique．Methods Samples from release solution of CDST at different time points were taken and tested according to stripping analysis method of sustained—release preparation and 45Ca determination of transmembrane influx technique．The small aortic rings from the heart of Wistar rats were prepared and equilibrated in the following solutions：physical saline solution(PSS)，45Ca-PSS+drug，and 45Ca—PSS+ KCl+drug one by one，then rinsed with cold EGTA solution，and finally digested．The radioactivity of the above samples was measured with liquid scintillation counter．Results The samples with extremely notable calcium antagonizing effect included CDST，Compound Danshen Tabler(CDT)，water—insoluble and water—soluble fractions of Danshen，total saponin from Sanqi(notoginseng)，tanshinoneⅡA,danshensu，protocatechuicaldehyde，ginsenoside Rbl，notoginsenoside R1，and borne01.CDST showed the significant calcium antagonizing effect 4 h later after being taken and extremely significant in 6h，while the self—prepared CDT showed the significant calcium antagonizing effect lh later and extremely significant in 2 h Conclusion water——insoluble and water——soluble It shows that CDST has notable calcium antagonizing effect，whereas fractions of Danshen and total saponin from Sanqi are active fractions of calcium antagonizing；tanshinone Ⅱ_A，danshensu，protocatechuic aldehyde,ginsenoside Rbl,notoginsenoside R1，and borneol are active ingredients.Compared to self—prepared CDT，the slower released effect of CDST iS available．Therefore。such a radiolabeled technique iS a feasible method for the comprehensive evaluation of calcium antagonizing effect as well as the sustained—release effect in vitro of Chinese medicine or natural medicine agents to treat vascular disorders．

国家自然科学基金青年科学基金资助项目(30000223)