摘要
目的建立细胞膜色谱法筛选吡罗昔康凝胶中的致敏组分,并进行致敏效应评价。方法采用类过敏反应受体MRGPRX2高表达细胞膜色谱模型对吡罗昔康凝胶的主成分和辅料进行致敏组分筛选;体外培养人肥大细胞LAD2,利用β-氨基己糖苷酶(β-Hex)及炎症因子释放实验验证所筛出组分的致敏效应;体外培养MRGPRX2高表达大鼠嗜碱性粒细胞性白血病细胞(RBL-MRGPRX2),验证吡罗昔康和月桂氮䓬酮(25、50、100、200μmol·L^(-1))对β-Hex释放率的影响;钙离子成像实验检验吡罗昔康和月桂氮䓬酮(200μmol·L^(-1))对MRGPRX2-HEK293细胞内钙离子的影响;检测吡罗昔康(5 mg·kg^(-1))、月桂氮䓬酮(2.5 mg·kg^(-1))对小鼠体温的影响。结果细胞膜色谱法从吡罗昔康凝胶中筛选出的致敏组分为吡罗昔康、月桂氮䓬酮、三乙醇胺和羟苯乙酯;与对照组比较,吡罗昔康和月桂氮䓬酮可显著促进β-Hex及肿瘤坏死因子α(TNF-α)的释放(P<0.05、0.01、0.001),其中月桂氮䓬酮作用呈浓度相关性,致敏效应最强;小鼠体温测定结果表明,与对照组比较,吡罗昔康和月桂氮䓬酮均可显著引发小鼠体温下降(P<0.05、0.01、0.001),且月桂氮䓬酮效应更强,与细胞水平结果一致。结论吡罗昔康凝胶中的吡罗昔康和月桂氮䓬酮具有引发类过敏反应的风险,其中月桂氮䓬酮的致敏效应更强。
Objective Cell membrane chromatography was established to screen the sensitizing components in piroxicam gel and evaluate their sensitizing effects.Methods A cell membrane chromatography model with high expression of the mast cell receptor MRGPRX2 was used to screen the main components and excipients of piroxicam gel for sensitizing components.Human mast cell line LAD2 was cultured in vitro,andβ-hexosaminidase(β-Hex)and inflammatory factor release experiments were used to verify the sensitizing effects of the screened components.RBL-MRGPRX2 cells with high expression of MRGPRX2 were cultured in vitro to verify the effects of piroxicam and laurindione(25,50,100,200μmol·L^(-1))on the release rate ofβ-Hex.Calcium ion imaging experiments were used to examine the effects of piroxicam and laurindione(200μmol·L^(-1))on intracellular calcium ions in MRGPRX2-HEK293 cells.The effects of piroxicam(5 mg·kg^(-1))and laurindione(2.5 mg·kg^(-1))on the body temperature of mice were detected.Results The sensitizing components screened from piroxicam gel by cell membrane chromatography were piroxicam,laurindione,triethanolamine and ethylparaben.Compared with the control group,piroxicam and laurindione could significantly promote the release ofβ-Hex and tumor necrosis factor(TNF)-α(P<0.05,0.01,0.001),and the sensitizing effect of laurindione was concentration-dependent and the strongest.The results of body temperature measurement in mice showed that compared with the control group,piroxicam and laurindione could significantly cause a decrease in body temperature in mice(P<0.05,0.01,0.001),and the effect of laurindione was stronger,which was consistent with the results at the cellular level.Conclusion Piroxicam and laurindione in piroxicam gel have the risk of causing anaphylactoid reactions,and the sensitizing effect of laurindione is stronger.
作者
李婕
庾莉菊
吴彦霖
王文林
陈华
LI Jie;YU Liju;WU Yanlin;WANG Wenlin;CHEN Hua(Key Laboratory of Chemical Drug Quality Research and Evaluation,National Medical Products Administration,National Institutes for Food and Drug Control,Beijing 102629,China;Shaanxi Institute for Food and Drug Control,Xi’an 710076,China)
出处
《药物评价研究》
北大核心
2025年第11期3143-3150,共8页
Drug Evaluation Research
基金
陕西省药品监管科学研究项目(2D/CMC分析方法在中药注液剂质量控制中的应用)。
