致敏RBL-2H3细胞在清开灵、痰热清中药注射剂过敏性评价中的应用  

Application of RBL-2H3 cells on allergenicity assessment of Qing kailing and Tanreqing Injections

作　　者：李黎明[1] 金若敏[1] 谢家骏[1] 姚广涛[1] 符胜光[1] 徐婷婷[1] 

机构地区：[1]上海中医药大学药物安全评价研究中心,上海201203

出　　处：《中草药》2015年第1期86-89,共4页Chinese Traditional and Herbal Drugs

基　　金：国家"重大新药创制"科技专项项目(2009ZX0902-002);国家"重大新药创制"科技专项项目(2011ZX09301-009)

摘　　要：目的探索采用血清抗体致敏的RBL-2H3细胞对清开灵、痰热清2种中药注射剂过敏性进行初步评价。方法以阳性药卵白蛋白(OVA)、清开灵注射液、痰热清注射液分别与Al(OH)3凝胶佐剂混合作为致敏源,Wistar大鼠sc致敏源制备抗体血清,采用放免法检测血清总Ig E水平。取抗体血清致敏RBL-2H3细胞,48 h后再以药物激发致敏细胞,检测细胞脱颗粒后上清液中β-氨基己糖苷酶释放率。另取大鼠进行被动皮肤过敏(PCA)实验,观察动物皮肤蓝斑阳性反应率。结果与对照组相比,OVA、清开灵、痰热清组大鼠抗体血清总Ig E水平著显升高(P<0.05、0.01)。致敏细胞脱颗粒结果显示:采用OVA和清开灵、痰热清注射液抗体血清致敏细胞,再经药物激发后RBL-2H3细胞上清液中β-氨基己糖苷酶释放率明显增加(P<0.05、0.01),与对照组比较,最大的相对释放倍数分别为3.7、1.53、1.98。大鼠PCA实验结果显示,各给药组大鼠蓝斑阳性反应最高百分率分别为100%、100%、86%。RBL-2H3细胞实验与PCA实验结果具有较好的一致性。结论血清抗体致敏RBL-2H3细胞模型可以用于对中药注射剂过敏性的初筛或评价。Objective To explore the allergenicity of two kinds of Chinese materia medica（CMM） injections, Qingkailing（QKL） and Tanreqing（TRQ） Injections, with serum antibody sensitized RBL-2H3 cells. Methods The antiserum was prepared by sc injection of allergen composed of ovalbumin（OVA）, QKL, or TRQ combined with aluminum hydroxide adjuvant respectively in Wistar rats. The total Ig E level in serum antibody was determined by radioimmunoassay. The RBL-2H3 cells were sensitized with serum antibodies, then stimulated by OVA, QKL, or TRQ after 48 h. The release rate of β-hexosaminidase in the supernatant was determined after the degranulation of sensitized RBL-2H3 cells. Passive cutaneous anaphylaxis（PCA） test was performed with rats, and the positive reaction rate with blue plaque in animal skin was observed. Results Compared with the control group, the total Ig E level in serum antibody was increased significantly in OVA, TRQ, and QKL groups（P〈0.05）. The degranulation test revealed that the release rate of β-hexosaminidase was significantly increased in the supernatant when the cells were incubated with the antiserum and then stimulated with OVA, QKL, and TRQ. Compared with the control group, the largest relative times of release were 3.7, 1.53, and 1.98, respectively. The results of PCA test showed that the highest percentage rates of positive reaction of blue plaque were 100%, 100%, and 86% respectively. The results of RBL-2H3 cell test and PCA test have good consistency. Conclusion The serum antibody sensitized RBL-2H3 cell model can be used for screening or assessing allergenicity of CMM injection.

关 键 词：致敏RBL-2H3细胞 清开灵注射液 痰热清注射液 过敏性 β-氨基己糖苷酶 

分 类 号：R285.5[医药卫生—中药学]