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目的研究多种注射用辅料刺激肥大细胞RBL-2H3脱颗粒的作用,建立和完善注射用辅料引发类过敏反应的筛选评价体系。方法以不同浓度的注射用辅料羟丙基-β-环糊精(HP-β-CD)、甲氧基聚乙二醇-聚乳酸共聚物40/60(mPEG-PDLLA40/60)、聚乙二醇400(PEG400)、聚乙二醇600(PEG600)、聚氧乙烯蓖麻油(CrEL)和吐温80处理RBL-2H3细胞,测定β-氨基己糖苷酶释放率,并通过MTT法进一步研究有脱颗粒作用的受试物对RBL-2H3细胞的毒性作用。结果 HP-β-CD、高浓度的mPEG-PDLLA40/60(40 mg/ml)和吐温80(3.0 mg/ml)均能直接刺激RBL-2H3细胞脱颗粒,呈浓度依赖性;PEG400、PEG600和CrEL在本研究的浓度范围内均不能直接刺激RBL-2H3细胞脱颗粒。HP-β-CD有明显的细胞毒性,但不呈浓度依赖性;mPEG-PDLLA40/60无细胞毒性;吐温80没有明显的细胞毒性,但呈浓度依赖性。结论 RBL-2H3细胞的β-氨基己糖苷酶释放试验方法灵敏、重复性好,其结果与临床报道有一定的相关性,提示其有评价注射用辅料类过敏反应的潜在应用价值。高浓度的吐温80直接刺激肥大细胞脱颗粒的作用可能是由其细胞毒性引起的。
Objective To study the effects of various injection excipients on the degranulation of mast cells RBL-2H3, and to establish and improve the screening evaluation system for allergic reactions induced by excipients in injection. Methods Different concentrations of hydroxypropyl-β-cyclodextrin (HP-β-CD), methoxypolyethylene glycol-polylactic acid copolymer 40/60 (mPEG-PDLLA40 / 60) RBL-2H3 cells were treated with PEG 400, PEG 600, CrEL and Tween 80. The release rate of β-hexosaminidase was determined and further studied by MTT Toxicity of test subjects with degranulation on RBL-2H3 cells. Results HP-β-CD, mPEG-PDLLA 40/60 (40 mg / ml) and Tween 80 (3.0 mg / ml) could directly stimulate the degranulation of RBL-2H3 cells in a dose- And CrEL did not directly stimulate the degranulation of RBL-2H3 cells in the concentration range of this study. HP-β-CD was significantly cytotoxic but not in a concentration-dependent manner; mPEG-PDLLA 40/60 was cytotoxic; and Tween 80 had no apparent cytotoxicity but in a concentration-dependent manner. Conclusion The method of beta-hexosaminidase release test in RBL-2H3 cells is sensitive and reproducible. The results are correlated with clinical reports, suggesting that it has potential value in the evaluation of allergic reactions in injectable adjuvants. The high concentration of Tween 80 directly stimulates mast cell degranulation may be caused by its cytotoxicity.