摘要
目的:利用基于发光细菌的Microtox测试技术,对国家药品不良反应监测中心数据库指引的高风险品种——参麦注射液的生物毒性实现定量化表征,初步探索导致其过敏反应的物质基础及临床意义,以期为参麦注射液的质量控制和不良反应风险预警提供新型、快速、灵敏的检测方法和技术平台。方法:(1)应用基于费氏弧菌CS234的Microtox测试体系对6个厂家共计15个批次的参麦注射液市售产品进行测试,得到相应的Microtox测试参数IC50。(2)采用高效液相色谱法对上述参麦注射液中三种代表性人参皂苷Re、Rg1及Rb1进行含量测定,分析IC50与三种人参皂苷及其总含量的相关性。(3)采用肥大细胞脱颗粒模型观察上述参麦注射液和人参皂苷Re对HMC-1细胞组胺及β-氨基己糖苷酶释放的影响。结果:(1)除A公司的参麦注射液无法计算IC50外(最高浓度受试样品的抑制率未达到50%),其余厂家的IC50从46.7%至72.3%不等,且存在显著性差异。(2)各厂家参麦注射液中人参皂苷Re及三种人参皂苷的总含量分别在0.07 mg/ml^0.14 mg/ml以及0.34 mg/ml^0.60 mg/ml范围内,非参数检验显示厂家间含量均存在显著性差异。经Pearson相关性分析显示,各厂家间IC50和人参皂苷Re含量之间存在显著的正相关关系(P=0.001),相关系数为0.823。(3)人参皂苷Re终浓度为0.10 mg/ml时,能显著增加HMC-1细胞上清液中组胺的含量。结论:本测试体系可用IC50定量表征参麦注射液的生物毒性,人参皂苷Re为显著影响IC50的物质之一,且Re在一定剂量下(终浓度0.10 mg/ml)对HMC-1细胞有显著的促组胺释放作用,提示参麦注射液中人参皂苷Re含量控制的必要性。基于费氏弧菌CS234的Microtox测试体系有望成为参麦注射液质量控制及不良反应风险预警的新型检测技术平台,以期最终为临床安全用药提供参考和保障。
Objective： To explore the material basis and its clinic significance so as to provide a new,rapid and sensitive detection platform for the quality control and early warnings,such as allergic reactions during clinical application of Shenmai injection. Methods：（ 1） IC50 of 15 batches of Shenmai injections purchased from 6 manufacturers were detected by microtox assay based on luminous bacterium Vibrio fischeri CS234.（ 2） According to chromatographic conditions in National Drug Standards of SM injection（ WS3-B-3428-98-2010 Z） issued by State Food and Drug Administration,contents of ginsenoside Re,Rg1 and Rb1 in Shenmai injection were assayed by high performance liquid chromatography（ HPLC）. Relationships between contents of ginsenoside Re,Rg1,Rb1,the total content of them and corresponding IC50 values of those Shenmai injections were analyzed by Pearson＇s correlation method,respectively.（ 3） Degranulation of HMC-1 cells caused by ginsenoside Re and the those Shenmai injections were also detected. Changes of cell morphology before and 1 hour after the administration were detected by the inverted microscope. Contents of histamine and β-hexosaminidase in the supernatant of cultured HMC-1 cells were detected by enzyme-linked immuno sorbent assay. Results：（ 1） The range of IC50 of Shenmai injections were from 46. 7 % to 72. 3%,except which purchased from SCSH Pharmaceutical Co.,LTD. One-way variance analysis（ ANOVA） showed that IC50 among different manufactories had a significant difference（ P〈0. 05）.（ 2） Contents of ginsenoside Re and total content of three ginsenosides were from 0. 07 mg/ml to 0. 14 mg/ml and 0. 34 mg/ml to 0. 60 mg/ml,respectively. Nonparametric tests showed that contents of ginsenoside Re and total contents of three had significant differences among different manufactories（ P〈0. 05）. Furthermore,there was a positive correlation between the content of ginsenoside Re and IC50（ r = 0. 823,P〈0. 01）.（ 3） In the degranulation test of mast cells
出处
《中药药理与临床》
CSCD
北大核心
2017年第6期36-41,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
重大新药创制科技重大专项项目(2015ZX09501004-001-005)
四川省科技计划项目(2016SZ0031)
四川省科技计划项目(2017TD0003)
