熊克朝1, 2,李泽君1, 2,李志刚4,张志伟4,张 纲5,吴纯启2,唐春萍1,丁日高2, 3,李振江4*,王全军2, 3*.参麦注射液致RBL-2H3细胞脱颗粒的研究[J].药物评价研究,2015,38():1-7
参麦注射液致RBL-2H3细胞脱颗粒的研究
Study on degranulation of RBL-2H3 cells induced by Shenmai Injection
投稿时间:2014-12-23  
DOI:
中文关键词:  参麦注射液;RBL-2H3细胞;类过敏反应
英文关键词:Shenmai Injection; RBL-2H3 cells; anaphylactoid reactions
基金项目:
作者单位
熊克朝1, 2,李泽君1, 2,李志刚4,张志伟4,张 纲5,吴纯启2,唐春萍1,丁日高2, 3,李振江4*,王全军2, 3* 1. 广东药学院中药学院,广东 广州 510006
2. 军事医学科医学院毒物药物研究所,国家北京药物安全评价研究中心,北京 100850
3. 抗毒药物与毒理学国家重点实验室(军事医学科学院),北京 100850
4. 河北神威药业有限公司,河北 三河 065201
5. 河北省中药注射剂工程技术研究中心,河北 石家庄 051430 
摘要点击次数: 1449
全文下载次数: 0
中文摘要:
      目的 探讨参麦注射液(SMI)对RBL-2H3细胞脱颗粒的影响及其原因。方法 以C48/80为工具药建立RBL-2H3细胞脱颗粒模型,检测不同浓度C48/80与RBL-2H3细胞作用不同时间后细胞β-己糖苷酶、类胰蛋白酶和组胺的释放率以及细胞活力,在细胞活力大于80%情况下,选择释放程度较高的指标和条件为优选考察指标和条件。将SMI和其溶剂(Tween- 80)原液等比稀释成不同浓度后与RBL-2H3细胞共同培养,通过中性红染色法观察细胞脱颗粒的形态学变化,分别用显色法和间接荧光法检测细胞上清的β-己糖苷酶和组胺释放率,采用细胞计数法(CCK-8)检测细胞的活力。结果 RBL-2H3细胞脱颗粒模型的最佳作用时间为30 min,最佳指标为β-己糖苷酶和组胺释放率。与空白组相比,SMI质量浓度低于13.3 g生药/L(3倍临床浓度)时,细胞中性红染色未见脱颗粒现象,组胺和β-己糖苷酶释放率亦无差异;而在Tween-80质量浓度为1.00 g/L时,SMI 40 g生药/L(9倍临床浓度)组和溶剂1.00 g/L组细胞中性红染色均可见脱颗粒现象,细胞上清组胺和β-己糖苷酶释放率亦明显增加。此外,CCK-8结果显示,与空白组相比,各浓度的SMI对细胞活力均无影响。结论 SMI低于3倍临床浓度无明显RBL-2H3细胞脱颗粒作用;而在9倍临床浓度能刺激细胞脱颗粒,这种脱颗粒作用可能与所含溶剂(Tween-80)有关,与其对RBL-2H3的细胞毒性作用无关。提示SMI在低于3倍临床浓度相对安全,在9倍临床浓度时有致类过敏反应的风险。
英文摘要:
      Objective To explore the effect on degranulation of RBL-2H3 cells and its reasons induced by Shenmai Injection (SMI). Methods Compound 48/80 (C48/80) was used to establish RBL-2H3 cells degranulation model, the release rates of β-hexosaminidase,Tryptase, and histamine and the cell viability of RBL-2H3 cells were detected after treated by different times at the different concentration of C48/80; The indicators and conditions which had a higher degree of release were selected as the preferred study indicators and conditions in the case of that the cell viability was more than 80 percents. RBL-2H3 cells were cultured with the geometric dilution at different concentrations of the stock solution of SMI and its solvent (Tween-80), the morphology of degranulation was observed by neutral red staining. In addition, the releasing rates of β-hexosaminidase and histamine in cell supernatants were detected by chromogenic assay and indirect fluorescence analysis, respectively; The viability of RBL-2H3 cells was tested by cell counting kits. Results The optimal condition of C48/80-induced RBL-2H3 cells degranulation test was 30 min and the best indicators were the release rates of β-hexosaminidase and histamine. When compared with the control group, the release rates of histamine and β-hexosaminidase were no difference and the degranulation by neutral red staining did not appear in SMI groups with the concentrations less than 13.3 g crude drug/L (3 times as much as the clinical concentration) while the release rates of histamine and β-hexosaminidase were significantly increased and the degranulation by neutral red staining appeared in SMI 40 g crude drug/L (9 times as the clinical concentration) group and SMI solvent group that both of them included 1.00 g/L Tween-80. In addition, the results of CCK-8 showed that there was no significant effect in different SMI doses groups on cell viability compared with the control group. Conclusion SMI at the concentration below 3 times as much as the clinical concentration has no significant degranulation of RBL-2H3 cells while at the concentration of 9 times as much as the clinical concentration shows the certain effect on inducing the degranulation of cells which may be related to its solvent that contains Tween-80 and shows no relationship with its toxic effects on the cells. It suggests that SMI is relatively safe at the concentration below 3 times as much as the clinical concentration while it has the risk of allergic reactions at the concentration of 9 times as much as the clinical concentration.
HTML  查看全文  查看/发表评论  下载PDF阅读器
关闭
您是第 2374789 位访客
版权所有 天津中草药杂志社 (天津市南开区鞍山西道308号 300193)
Address: 308# An-shan West Road, Nankai District, Tianjin 300193, China
Tel: +86-22-27474913; 23006822 Fax: +86-022-23006822 E-mail:zcy@tiprpress.com
津备案:津ICP备13000267号 互联网药品信息服务资格证书编号:(津)-非经营性-2015-0031