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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 Qingbailing and Tanreqing Injection with RBL-2H3 cells sensitized by serum antibody. Methods The positive serum ovalbumin (OVA), Qingkailing injection and Tanreqing injection were mixed with Al (OH) 3 gel adjuvant respectively, Radioimmunoassay for serum total Ig E levels. Antibody sera were used to sensitize RBL-2H3 cells. After 48 h, the cells were sensitized with drugs and the release rate of β-hexosaminidase in the supernatant was measured. Another rat for passive skin allergy (PCA) experiments to observe the positive reaction rate of animal skin blue spot. Results Compared with the control group, the levels of total serum IgE in OVA, Qingkailing and Tanreqing groups were significantly increased (P <0.05, 0.01). The results of desensitization of sensitized cells showed that the release rate of β-hexosaminidase in the supernatant of RBL-2H3 cells was significantly increased by OVA and Qingkailing and Tanreqing injection serum-sensitized cells ( P <0.05,0.01), compared with the control group, the maximum relative release multiple were 3.7,1.53,1.98. The results of rat PCA showed that the highest percentage of positive plaques in rats in each administration group was 100%, 100% and 86%, respectively. RBL-2H3 cells and PCA experimental results have good agreement. Conclusion Serum antibody-sensitized RBL-2H3 cell model can be used to screen or evaluate the sensitivity of traditional Chinese medicine injection.