目的:建立大鼠体外肥大细胞以及RBL-2H3细胞脱颗粒试验的方法,评估2种细胞脱颗粒方法对中药注射剂(traditional Chinese medicine injections,TCMI)引起脱颗粒的可行性。方法:提取大鼠腹腔肥大细胞(rat peritoneal mast cell,RPMC),优...目的:建立大鼠体外肥大细胞以及RBL-2H3细胞脱颗粒试验的方法,评估2种细胞脱颗粒方法对中药注射剂(traditional Chinese medicine injections,TCMI)引起脱颗粒的可行性。方法:提取大鼠腹腔肥大细胞(rat peritoneal mast cell,RPMC),优化实验条件,用阳性药物C48/80和各种TCMI等与其共培养,通过甲苯胺蓝染色法计数腹腔MC脱颗粒率;通过RBL-2H3细胞与TCMI共同培养,底物显色法检测β-氨基己糖苷酶释放率。结果:阳性对照品C48/80的浓度与RPMC脱颗粒有一定的量效关系;参麦、冠心宁、痰热清、清开灵和生脉5种TCMI能显著引起细胞脱颗粒,并与药物浓度有一定的正相关性;RBL-2H3细胞脱颗粒方法中参麦、黄芪、银杏、痰热清、丹香冠心、清开灵和生脉7种TCMI能是β-氨基己糖苷酶释放率升高。结论:一些TCMI能引起体外RPMC和RBL-2H3细胞脱颗粒,2种细胞脱颗粒方法在评价一些TCMI可能引起类过敏反应方面具有一定的互补性与可行性。展开更多
目的通过建立体外抑制肥大细胞脱颗粒的药效学模型,探讨丹皮酚抗Ⅰ型变态反应作用机制。方法用MTT比色法检测不同浓度的丹皮酚对RBL-2H3肥大细胞增殖的影响。通过ELISA法,明确体外RBL-2H3细胞活化分泌组胺及TNF-α动力学特征。检测不同...目的通过建立体外抑制肥大细胞脱颗粒的药效学模型,探讨丹皮酚抗Ⅰ型变态反应作用机制。方法用MTT比色法检测不同浓度的丹皮酚对RBL-2H3肥大细胞增殖的影响。通过ELISA法,明确体外RBL-2H3细胞活化分泌组胺及TNF-α动力学特征。检测不同浓度药物对RBL-2H3肥大细胞分泌组胺及TNF-α的影响。结果丹皮酚对RBL-2H3细胞增殖有抑制作用,IC50值为0.22 mg/mL。RBL-2H3细胞活化脱颗粒,分泌组胺的释放率在前30 m in上升较快,30 m in后上升进入平台期;TNF-α分泌的量在1h达高峰。丹皮酚抑制RBL-2H3细胞释放组胺和TNF-α作用不弱于色甘酸钠0.1,0.5 mg/mL。丹皮酚浓度的对数与其对RBL-2H3细胞释放组胺和TNF-α的抑制率呈线性相关(P=0.000<0.01)。结论丹皮酚呈剂量依赖性抑制RBL-2H3细胞分泌组胺和TNF-α。展开更多
The present study aimed at developing a natural compound with anti-allergic effect and stability under latex glove manufacturing conditions and investigating whether its anti-allergic effect is maintained after its ad...The present study aimed at developing a natural compound with anti-allergic effect and stability under latex glove manufacturing conditions and investigating whether its anti-allergic effect is maintained after its addition into the latex. The effects of nine natural compounds on growth of the RBL-2H3 cells and mouse primary spleen lymphocytes were determined using MTT assay. The compounds included glycyrrhizin, osthole, tetrandrine, tea polyphenol, catechin, arctigenin, oleanolic acid, baicalin and oxymatrine. An ELISA assay was used for the in vitro anti-type I/IV allergy screening; in this process β-hexosaminidase, histamine, and IL-4 released from RBL-2H3 cell lines and IFN-γ and IL-2 released from mouse primary spleen lymphocytes were taken as screening indices. The physical stability of eight natural compounds and the dissolubility of arctigenin, selected based on the in vitro pharnacodynamaic screening and the stability evaluation, were detected by HPLC. The in vivo pharmacodynamic confirmation of arctigenin and final latex product was evaluated with a passive cutaneous anaphylaxis(PCA) model and an allergen-specific skin response model. Nine natural compounds showed minor growth inhibition on RBL-2H3 cells and mouse primary spleen lymphocytes. Baicalin and arctigenin had the best anti-type I and IV allergic effects among the natural compounds based on the in vitro pharmacodynamic screening. Arctigenin and catechin had the best physical stability under different manufacturing conditions. Arctigenin was the selected for further evaluation and proven to have anti-type I and IV allergic effects in vivo in a dose-dependent manner. The final product of the arctigenin-containing latex glove had anti-type I and IV allergic effects in vivo which were mainly attributed to arctigenin as proved from the dissolubility results. Arctigenin showed anti-type I and IV allergic effects in vitro and in vivo, with a good stability under latex glove manufacturing conditions, and a persistent anti-allergic effect after being 展开更多
文摘目的:建立大鼠体外肥大细胞以及RBL-2H3细胞脱颗粒试验的方法,评估2种细胞脱颗粒方法对中药注射剂(traditional Chinese medicine injections,TCMI)引起脱颗粒的可行性。方法:提取大鼠腹腔肥大细胞(rat peritoneal mast cell,RPMC),优化实验条件,用阳性药物C48/80和各种TCMI等与其共培养,通过甲苯胺蓝染色法计数腹腔MC脱颗粒率;通过RBL-2H3细胞与TCMI共同培养,底物显色法检测β-氨基己糖苷酶释放率。结果:阳性对照品C48/80的浓度与RPMC脱颗粒有一定的量效关系;参麦、冠心宁、痰热清、清开灵和生脉5种TCMI能显著引起细胞脱颗粒,并与药物浓度有一定的正相关性;RBL-2H3细胞脱颗粒方法中参麦、黄芪、银杏、痰热清、丹香冠心、清开灵和生脉7种TCMI能是β-氨基己糖苷酶释放率升高。结论:一些TCMI能引起体外RPMC和RBL-2H3细胞脱颗粒,2种细胞脱颗粒方法在评价一些TCMI可能引起类过敏反应方面具有一定的互补性与可行性。
文摘目的通过建立体外抑制肥大细胞脱颗粒的药效学模型,探讨丹皮酚抗Ⅰ型变态反应作用机制。方法用MTT比色法检测不同浓度的丹皮酚对RBL-2H3肥大细胞增殖的影响。通过ELISA法,明确体外RBL-2H3细胞活化分泌组胺及TNF-α动力学特征。检测不同浓度药物对RBL-2H3肥大细胞分泌组胺及TNF-α的影响。结果丹皮酚对RBL-2H3细胞增殖有抑制作用,IC50值为0.22 mg/mL。RBL-2H3细胞活化脱颗粒,分泌组胺的释放率在前30 m in上升较快,30 m in后上升进入平台期;TNF-α分泌的量在1h达高峰。丹皮酚抑制RBL-2H3细胞释放组胺和TNF-α作用不弱于色甘酸钠0.1,0.5 mg/mL。丹皮酚浓度的对数与其对RBL-2H3细胞释放组胺和TNF-α的抑制率呈线性相关(P=0.000<0.01)。结论丹皮酚呈剂量依赖性抑制RBL-2H3细胞分泌组胺和TNF-α。
基金supported by National Science and Technology Infrastructure Program of China(No.2012BAI30B001)Technological Innovation Team of Jiangsu Higher Education+3 种基金National Natural Science Foundation of China(Nos.3087315630672524)New Century Excellent Talents in University(NCET-07-0851)Mega-projects of Science Research for the 12th Five-Year Plan of China(No.2011AX09401-007)
文摘The present study aimed at developing a natural compound with anti-allergic effect and stability under latex glove manufacturing conditions and investigating whether its anti-allergic effect is maintained after its addition into the latex. The effects of nine natural compounds on growth of the RBL-2H3 cells and mouse primary spleen lymphocytes were determined using MTT assay. The compounds included glycyrrhizin, osthole, tetrandrine, tea polyphenol, catechin, arctigenin, oleanolic acid, baicalin and oxymatrine. An ELISA assay was used for the in vitro anti-type I/IV allergy screening; in this process β-hexosaminidase, histamine, and IL-4 released from RBL-2H3 cell lines and IFN-γ and IL-2 released from mouse primary spleen lymphocytes were taken as screening indices. The physical stability of eight natural compounds and the dissolubility of arctigenin, selected based on the in vitro pharnacodynamaic screening and the stability evaluation, were detected by HPLC. The in vivo pharmacodynamic confirmation of arctigenin and final latex product was evaluated with a passive cutaneous anaphylaxis(PCA) model and an allergen-specific skin response model. Nine natural compounds showed minor growth inhibition on RBL-2H3 cells and mouse primary spleen lymphocytes. Baicalin and arctigenin had the best anti-type I and IV allergic effects among the natural compounds based on the in vitro pharmacodynamic screening. Arctigenin and catechin had the best physical stability under different manufacturing conditions. Arctigenin was the selected for further evaluation and proven to have anti-type I and IV allergic effects in vivo in a dose-dependent manner. The final product of the arctigenin-containing latex glove had anti-type I and IV allergic effects in vivo which were mainly attributed to arctigenin as proved from the dissolubility results. Arctigenin showed anti-type I and IV allergic effects in vitro and in vivo, with a good stability under latex glove manufacturing conditions, and a persistent anti-allergic effect after being
