陈勇灵,蔡犇,张巍,解卓奇,黄维,潘丽,卢敏
CHEN Yong-ling,CAI Ben,ZHANG Wei,XIE Zhuo-qi,HUANG Wei,PAN Li,LU Min

• 1:浙江永宁药业股份有限公司

摘要(Abstract)：

建立RBL-2H3细胞脱颗粒模型及小鼠急性类过敏反应模型的注射用红花黄色素类过敏反应筛查方法,评价不同批次注射用红花黄色素的致类过敏性。采用体外细胞模型,以各批次注射用红花黄色素为药物组,无血清MEM培养基为阴性对照,30 mg·L-1C48/80为阳性对照分别刺激细胞1 h,吸取上清液,以中性红染色法,以发生脱颗粒的阳性细胞率为指标检测注射用红花黄色素对RBL-2H3细胞脱颗粒的影响;建立体内模型以验证细胞实验结果,采用小鼠急性类过敏反应模型,ELISA法检测血浆组胺含量,以血浆组胺含量为检测指标筛查注射用红花黄色素在动物水平的致类过敏性。结果表明,中性红染色实验结果显示阳性对照C48/80能明显引起细胞脱颗粒,大部分细胞深染,而不同批次注射用红花黄色素阳性细胞率与阳性对照有显著性差异;小鼠急性类过敏反应模型中,通过血浆组胺含量的检测,阳性对照C48/80可显著增加小鼠血浆中组胺的含量,而各批次注射用红花黄色素均未引起血浆组胺明显升高(P<0. 000 1)。类过敏反应机制较为复杂,该研究主要基于肥大细胞脱颗粒释放组胺等活性物质,从体内体外2个方面,未检测到注射用红花黄色素引起RBL-2H3细胞发生明显的脱颗粒反应,也未使小鼠血浆组胺水平显著上升。
This paper was aimed to establish screening methods of anaphylactoid reaction caused by safflower yellow for injection based on RBL-2 H3 cell degranulation model and mice model for acute anaphylactoid reaction,and evaluate the hypersensitivity caused by safflower yellow for injection from different batches. An in vitro cell model was used to keep the cells stimulated for an hour with different batches of safflower yellow for injection as the drug group,serum-free MEM medium as negative control group and 30 mg·L-1 C48/80 as positive control group respectively. The supernatant was then absorbed,and neutral red staining technique was used to detect the effect of safflower yellow injection on the degranulation of RBL-2 H3 cells with the positive cell rate of degranulation as the indicator.An in vivo model was established to validate the experimental results,and mice model for acute anaphylactoid reaction and ELISA method were adopted to detect the plasma histamine content,and screen the hypersensitivity caused by safflower yellow for injection at the animal level by using plasma histamine content as a test index. The results of the neutral red staining experiments showed that the positive control C48/80 could cause cell degranulation,and most of the cells were deeply stained. There was significant difference in positive cell rate between different batches of safflower yellow and positive control group. In the mice model for acute anaphylactoid reaction,it was found that the positive control C48/80 significantly increased the histamine content in the plasma of mice,while the safflower yellow in each batch did not cause a significant increase in plasma histamine( P<0. 000 1). The mechanism of anaphylactoid reaction is relatively complicated. This study was mainly based on the release of histamine and other active substances by degranulation of mast cells. No significant degranulation reaction of RBL-2 H3 cells induced by safflower yellow for injection was detected,nor was the plasma histamine level significantly increased in mice from the in vitro and in vivo aspects.

关键词(KeyWords)： 红花黄色素;脱颗粒;MTT;组胺;类过敏
safflower yellow;degranulation;MTT;histamine;anaphylactoid

Abstract:

Keywords:

基金项目(Foundation): 浙江省重点研发计划项目(2019C03067)

作者(Author): 陈勇灵,蔡犇,张巍,解卓奇,黄维,潘丽,卢敏
CHEN Yong-ling,CAI Ben,ZHANG Wei,XIE Zhuo-qi,HUANG Wei,PAN Li,LU Min

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