BN大鼠和RBL-2H3细胞体内外实验评价丹红注射液致敏性风险

Evaluation of the sensitization risk of Danhong Injection based on in vivo and in vitro experiments using BN rats and RBL-2H3 cells

关键词:  丹红注射液  类过敏  BN大鼠  脱颗粒

Key words:  Danhong Injection  pseudo-allergy  BN rat  degranulation

摘要:

[目的] 研究通过体内外实验评估丹红注射液的致敏性风险。[方法] 丹红注射液与临床常用溶媒配伍后进行理化性质相关检测。体外实验中，大鼠嗜碱性粒细胞白血病细胞（RBL-2H3）给药分组设置为阴性对照组、阳性对照组（C48/80组）、丹红注射液（20、30、50 μL/mL）组分别检测细胞活力、甲苯胺蓝染色、脱颗粒标志物[免疫球蛋白E（IgE）、组胺（His）、β氨基已糖苷酶（β-Hex）、类胰蛋白酶（TPS）]。采用BN大鼠致敏模型（阴性对照组、阳性对照组、丹红氯化钠组、丹红葡萄糖组），通过双次激发实验观察过敏反应，取血及肺组织进行病理分析。[结果] 丹红注射液与溶媒配伍后理化性质稳定；高剂量丹红注射液（50 μL/mL）（P<0.05）显著提升RBL-2H3细胞His释放量。体内实验未发现BN大鼠出现显著过敏反应。[结论] 丹红注射液在常规剂量下未表现显著致敏性。

Abstract:

[Objective] This study aimed to evaluate the sensitization risk of Danhong Injection through in vivo and in vitro experiments. [Methods] The physicochemical properties of Danhong Injection were tested after compatibility with commonly used clinical solvents. For the in vitro experiment，RBL-2H3 cells were allocated to a negative control group，a positive control group（C48/80），and Danhong Injection groups（20，30 and 50 μL/mL）. Cell viability，toluidine blue staining，and the levels of degranulation markers（immunoglobulin E（IgE），histamine（His），β-hexosaminidase（β-Hex），and tryptase（TPS）） were assessed. A BN rat sensitization model was established， including normal saline control，ovalbumin positive control，Danhong Injection-sodium chloride injection compatibility，and Danhong Injection-glucose injection compatibility groups. Allergic reactions were observed following a double-challenge protocol. Blood and lung tissues were collected for histopathological analysis. [Results] Danhong Injection exhibited stable physicochemical properties after compatibility. In RBL-2H3 cells，the high dose（50 μL/mL） of Danhong Injection significantly increased histamine release（P<0.05）. However，no significant allergic reactions were observed in BN rats in vivo. [Conclusion] Danhong Injection did not demonstrate significant sensitization risk at conventional clinical doses.