摘要
目的检测双黄连注射剂中1,5-二咖啡酰奎宁酸致敏家兔后血清中特异性抗体效价。方法双黄连注射剂中疑似小分子半抗原物质1,5-二咖啡酰奎宁酸与正常家兔血清体外孵育制备完全抗原,免疫家兔获得特异性抗体。用酶联免疫吸附试验(ELISA)检测抗血清的抗体效价。间接竞争ELISA测定血清特异性并绘制抑制曲线。采用兔免疫球蛋白E(IgE)ELISA试剂盒检测各组家兔抗血清中IgE抗体的含量。结果1,5-二咖啡酰奎宁酸浓度分别为0.25 mg/ml、0.50 mg/ml、1.00 mg/ml、2.00 mg/ml、4.00 mg/ml时效价[吸光度(A值)]对应的分别为0.364±0.020、0.334±0.010、0.322±0.020、0.332±0.010、0.298±0.020,均高于阴性对照组A值(0.095±0.010)。得到回归方程I=0.1952 lgC+0.2734,R2=0.9958,半数抑制浓度(IC50)=33.494 mg/ml。家兔体内产生了外源性IgE型抗体。结论在本实验条件下双黄连注射剂中疑似半抗原物质1,5-二咖啡酰奎宁酸具有致敏性。
Objective To determine the specific antibodies in the serum of rabbits sensitized by 1,5-di caffeoylquinic acid in Shuanghuanglian Injection by ELISA.Methods The suspected small molecule hapten substance 1,5-dicaffeoylquinic acid in Shuanghuanglian Injection was incubated with normal rabbit serum in vitro to prepare a complete antigen,which was then used to immunize rabbits to obtain specific antibodies.The antibody titer of antiserum was detected by ELISA method.The serum specificity was determined by indirect competitive ELISA and the inhibition curve was plotted.The content of IgE antibodies in the antiserum of rabbits in each group was detected using a rabbit Immunoglobulin E(IgE)ELISA kit.Results The A values of 1,5-dicaffeoylquinic acid of 0.25 mg/ml,0.50 mg/ml,1.00 mg/ml,2.00 mg/ml,and 4.00 mg/ml were(0.364±0.020),(0.334±0.010),(0.322±0.020),(0.332±0.010),and(0.298±0.020),respectively,which were higher than the A value of the negative control group(0.095±0.010).The regression equation obtained was I=0.1952 lgC+0.2734,with R2=0.9958,and the half maximal inhibitory concentration(IC50)was 33.494 mg/ml.Exogenous IgE-type antibodies were obviously produced in rabbits.Conclusion The suspected hapten 1,5-dicaffeoylquinic acid in Shuanghuanglian Injection is allergenic under the experimental conditions.
作者
邓颖颖
闫瑞
徐丽婷
李坤
折宝霞
史珍珍
Deng Yingying;Yan Rui;Xu Liting;Li Kun;She BaoxiaShi Zhenzhen(Department of Pharmacy,Xi′an International Medical Center Hospital,Xi′an,Shaanxi 710100,China)
出处
《中国药物与临床》
CAS
2024年第19期1289-1292,共4页
Chinese Remedies & Clinics
关键词
1
5-二咖啡酰奎宁酸
双黄连注射剂
酶联免疫吸附测定
1,5-dicaffeoyl quinic acid
Shuang Huang Lian Zhu She Ji
Enzyme-linked immunosorbent assay
作者简介
通信作者:闫瑞,Email:429587459@qq.com。
