摘要
目的利用酶联免疫吸附测定法(ELISA)法检测双黄连注射剂中黄芩素致敏家兔后血清中特异性抗体的效价。方法将正常家兔血清与双黄连注射剂中疑似小分子半抗原物质黄芩素在体外一起孵育从而制备所需完全抗原,随后,利用这些完全抗原免疫家兔以期获得特异性抗体。运用ELISA法检测所得抗血清的抗体效价。通过间接竞争ELISA法,进一步测定血清特异性并绘制相关浓度抑制曲线。此外,采用兔免疫球蛋白E(IgE)ELISA试剂盒检测各组家兔抗血清中相对应的IgE抗体含量。结果与阴性对照组比较黄芩素的抗体效价(OD)值是其2倍,表明其具有潜在的致敏性。通过整理数据,绘制抑制率曲线得到回归方程I=0.138lgC+0.402,R^(2)=0.959,显示出良好的拟合度。半数抑制浓度IC_(50)=5.178 mg·mL^(-1)。此外,本实验结果明显表明外源性IgE型抗体已经在家兔体内产生,从而进一步证实了黄芩素的致敏性。结论在本实验所模拟家兔致敏全过程结果中表明黄芩素具有明显的致敏性。这一发现对于双黄连注射剂的安全使用具有重要的指导意义,也为相关药物的研发和生产提供了有价值的参考。
OBJECTIVE To detect the titer of specific antibodies in baicali n-sensitized rabbit serum of ShuangHuangLian injection by enzyme-linke d immunosorbent assay(ELISA).METHODS Normal rabbit serum was incubated with baicalin,a suspected small molecule hapten substance in ShuangHuangLian injection,in vitro to prepare the required complete antigens,and then th e rabbits were immunized with these complete antigens in order to obtain specifi c antibodies.The antibody titer of the antiserum was detected by ELISA.By indi rect competitive ELISA,serum specificity was further determined and the relevan t concentration inhibition curves were plotted.In addition,rabbit immunoglobul in E(IgE)ELISA kit was used to detect the corresponding IgE antibody content i n the antiserum of rabbits in each group.RESULTS The antibody titer(OD)value of baicalein was twice that of the negative control group,indi cating its potential allergenicity.By collating the data and plotting the inhib ition rate curve,the regression equations I=0.138lgC+0.402 and R^(2)=0.959 we re obtained,showing a good fit.The half inhibitory concentration was IC_(50)=5.178 mg·mL^(-1).In addition,the results of this experiment clearly show ed that exogenous IgE antibodies had been produced in rabbits,which further con firmed the sensitization of baicalin.CONCLUSION The results of the whole process of rabbit sensitization simulated in this laboratory showed t hat baicalein had obvious allergenicity.This finding has important guiding sign ificance for the safe use of ShuangHuangLian injection,and also provides a valu able reference for the development and production of related drugs.
作者
邓颖颖
徐丽婷
李坤
王皎
折宝霞
DENG Ying-ying;XU Li-ting;LI Kun;WANG Jiao;SHE Bao-xia(Department of Pharmacy,Xi′an International Medical Center Hospital,Xi′an 710100,China)
出处
《海峡药学》
2024年第9期1-3,共3页
Strait Pharmaceutical Journal
基金
青年课题(项目编号:2022QN30)。
