丹红注射液在体外抑制血小板聚集的生物活性测定法研究

Research on bioassay method of Danhong injection in inhibiting platelet aggregation in vitro

目的:以丹红注射液的抗血小板聚集活性为切入点,旨在建立丹红注射液的生物活性测定法,以期量化丹红注射液的药理作用,并优化现有的中药注射液质量评价体系。方法:以诱导剂为花生四烯酸(arachidonic acid),丹红注射液剂间比1∶0.8,血小板数500×10^(9)个·L^(-1),在体外抑制血小板聚集的生物作用为试验系,实施量效关系考察与效价定义,而后通过量反应平行线法计算效价,建立丹红注射液的生物活性测定法,同时进行方法学考察验证该方法的可行性与适用性。结果:丹红注射液在64%~100%的浓度范围内呈现较显著的量效关系,且由效价定义得到的丹红注射液标准品效价为1350 U·mL^(-1);其余7个不同批次的供试品效价分别为1320.6、1397.4、1407.5、1445.2、1278.4及1234.6 U·mL^(-1)。结论:基于抗血小板聚集活性建立的丹红注射液生物活性测定法操作简便,精确度高,同时以效价测定的方式量化了丹红注射液的关键活性,可为现有中药质量控制方法的改进与扩充提供借鉴。

Objective:Taking the anti-platelet aggregation activity of Danhong injection as the breakthrough point,to establish a bioassay method for Danhong injection,in order to quantify the pharmacological effect of Danhong injection and optimize the existing quality evaluation system of traditional Chinese medicine injection.Methods:Arachidonic acid was used as the inducer,the dose ratio of Danhong injection was 1∶0.8,and the platelet count was 500×10^(9)L^(-1).The inhibition of platelet aggregation in vitro by Danhong injection was used as the experimental system,to examine the dose-effect relationship and define the valence,and then thevalences were calculated by the dose-response parallel line method,and the bioassay method of Danhong injection was developed.At the same time,the feasibility and applicability of the method were verified by methodological investigation.Results:The results showed that Danhong injection displayed a significant dose-effect relationship within the concentration range of 64%-100%,and the valence of Danhong injection standard obtained from the valence definition was 1350 U·mL^(-1).The valences of the remaining seven different batches of the test articles were 1320.6,1397.4,1407.5,1445.2,1278.4 and 1234.6 U·mL^(-1),respectively.Conclusion:The established bioassay method of Danhong injection based on anti-platelet aggregation activity is simple to operate and highly accurate,while quantifying the key activity of Danhong injection by valence determination,which provides a reference for the improvement and expansion of current quality control methods for traditional Chinese medicine.