白芍总苷对人肥大细胞组胺释放活性的影响
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摘要
目的观察白芍总苷(total glucosides of paeony,TGP)对人肥大细胞组胺释放的影响,探讨TGP治疗慢性荨麻疹等变态反应性疾病的机制。方法取正常成人包皮,采用酶消化法分离肥大细胞,甲苯胺蓝染色鉴定肥大细胞。不同浓度TGP(100、200、400、800、1600μg/ml)作用肥大细胞24 h后,采用细胞计数试剂盒(cell counting kit 8,CCK-8)检测TGP对肥大细胞毒性的影响。在抗Ig E抗体激活肥大细胞后,采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测不同TGP浓度(200、400、600、800μg/ml)对肥大细胞组胺释放的影响。结果经甲苯胺蓝染色法鉴定后培养出来的细胞为肥大细胞。TGP的浓度在100、200、400、600、800、1600μg/ml时,对正常肥大细胞均无明显毒性。抗Ig E抗体激活肥大细胞后,200、400、600、800μg/ml TGP可抑制肥大细胞组胺的释放。结论采用此法成功分离出人肥大细胞,TGP在一定浓度范围内对人正常肥大细胞无明显细胞毒性,但能显著抑制人肥大细胞组胺的释放。
Objective To observe the effect of total glucosides of paeony( TGP) on histamine release from human mast cells,and explore the mechanism of TGP in the treatment of allergic diseases such as chronic urticaria. Methods Mast cells were isolated from normal adult foreskin by enzyme digestion,and identified by toluidine blue staining. The cell toxicity of TGP to normal mast cells were detected by cell counting kit 8( CCK-8) at 24 hours after treating cells with different concentrations( 100,200,400,600,800,1600 μg/ml) of TGP. The activities of histamine release from mast cells at the different concentrations( 200,400,600,800 μg/ml) of TGP were measured by enzyme linked immunosorbent assay( ELISA) after the activation of mast cells by anti-IgE antibody. Results The isolated cells were identified as human mast cells by toluidine blue staining. TGP had no obvious toxicity to normal mast cells at the concentrations of 100,200,400,600,800,1600 μg/ml. The histamine release from mast cells were inhibited by TGP at the concentrations of 200,400,600,800 μg/ml after the activation of mast cells by anti-IgE antibody. Conclusion The human mast cell is succsessfully isolated and identified. TGP has no obvious cytotoxicity to human normal mast cells in a certain concentration range,but it can significantly inhibit the release of histamine from human mast cells.
Objective To observe the effect of total glucosides of paeony( TGP) on histamine release from human mast cells,and explore the mechanism of TGP in the treatment of allergic diseases such as chronic urticaria. Methods Mast cells were isolated from normal adult foreskin by enzyme digestion,and identified by toluidine blue staining. The cell toxicity of TGP to normal mast cells were detected by cell counting kit 8( CCK-8) at 24 hours after treating cells with different concentrations( 100,200,400,600,800,1600 μg/ml) of TGP. The activities of histamine release from mast cells at the different concentrations( 200,400,600,800 μg/ml) of TGP were measured by enzyme linked immunosorbent assay( ELISA) after the activation of mast cells by anti-IgE antibody. Results The isolated cells were identified as human mast cells by toluidine blue staining. TGP had no obvious toxicity to normal mast cells at the concentrations of 100,200,400,600,800,1600 μg/ml. The histamine release from mast cells were inhibited by TGP at the concentrations of 200,400,600,800 μg/ml after the activation of mast cells by anti-IgE antibody. Conclusion The human mast cell is succsessfully isolated and identified. TGP has no obvious cytotoxicity to human normal mast cells in a certain concentration range,but it can significantly inhibit the release of histamine from human mast cells.
引文
[1]赵洪伟,崔银锋,姜京植.芝麻素通过PKCα/NF-κB信号途径抑制肥大细胞活化[J].中国免疫学杂志,2018,34(2):167-171
[2]Kalesnokoff J,Galli SJ.Newdevelopment in mast cell biology[J].Nat Immunol,2008,9(11):1215-1223
[3]Abraham SN,St John AL.Mast cell-orchestrated immunity to pathogens[J].Nat Rev Immunol,2010,10(6):440-452
[4]Guillén D,Fiandor-Román A,Caballero T,et al.Urticaria canused by ingestion of pasta and bread containing buckwheat flour[J].J Investig Allergol Clin Immunol,2013,23(3):206-207
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[7]张宇虹,郭在培,焦晓燕,等.白芍总苷作用机理及其在皮肤科的应用[J].中国麻风皮肤病杂志,2012,28(2):109-111
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[10]谢华,何韶衡,傅意玲.蛋白酶激活受体-2激动剂对人皮肤和扁桃体肥大细胞释放组胺和类胰蛋白酶的影响[J].第四军医大学学报,2005,26(7):599-602
[11]路敏敏,金世禄,刘宝珍,等.肥大细胞在胃癌及癌前病变组织中的分布及其临床意义[J].川北医学院学报,2015,30(4):442-445
[12]代渊,任波,陈欢,等.肤乐颗粒对大鼠颅骨肥大细胞脱颗粒作用的拮抗效应及机制[J].中国老年学杂志,2017,37(16):3911-3914
[13]王珊,赵作涛,王誉涵,等.肥大细胞在金黄色葡萄球菌肠毒素B诱导小鼠特应性皮炎样炎症反应中的作用[J].中华皮肤科杂志,2018,51(1):26-30
[14]唐慧,王朵勤,沈燕芸,等.屋尘螨变应原对人肥大细胞脱颗粒的影响[J].上海医药,2016,37(9):10-13
[15]James C,Bernstein JA.Current and future therapies for thetreatment of histamine-induced angioedema[J].Expert Opin Pharmacother,2017,18(3):253-262
[16]席海英,霍月红.依巴斯汀联合白芍总苷、雷公藤多甙治疗慢性荨麻疹的研究[J].医学信息,2016,29(28):64-65
[17]陈文慧,刘乐,李伯埙.白芍总苷联合苯磺贝他斯汀治疗慢性荨麻疹疗效观察[J].中国中西医结合皮肤性病学杂志,2016,15(6):355-357
[18]王小艳,陈星宇,张洪英.白芍总苷在皮肤科的应用进展[J].中国中西医结合皮肤性病学杂志,2015,4(4):262-264
[2]Kalesnokoff J,Galli SJ.Newdevelopment in mast cell biology[J].Nat Immunol,2008,9(11):1215-1223
[3]Abraham SN,St John AL.Mast cell-orchestrated immunity to pathogens[J].Nat Rev Immunol,2010,10(6):440-452
[4]Guillén D,Fiandor-Román A,Caballero T,et al.Urticaria canused by ingestion of pasta and bread containing buckwheat flour[J].J Investig Allergol Clin Immunol,2013,23(3):206-207
[5]赵辨.中国临床皮肤病学[M].2版.南京:江苏凤凰科学技术出版社,2017:783-785
[6]王海坤.白芍总苷在风湿免疫性疾病中的临床应用[J].中国实用医药,2016,11(10):280-281
[7]张宇虹,郭在培,焦晓燕,等.白芍总苷作用机理及其在皮肤科的应用[J].中国麻风皮肤病杂志,2012,28(2):109-111
[8]孙蔚凌,毕志刚.慢性荨麻疹患者血清组胺释放活性检测[J].中华皮肤科杂志,2004,37(6):351-353
[9]He SH,Xie H,He YS.Induction of tryptase and histamine release from human colon mast cells by Ig E dependent or independent mechanisms[J].World J Gastroenterol,2004,10(3):319-322
[10]谢华,何韶衡,傅意玲.蛋白酶激活受体-2激动剂对人皮肤和扁桃体肥大细胞释放组胺和类胰蛋白酶的影响[J].第四军医大学学报,2005,26(7):599-602
[11]路敏敏,金世禄,刘宝珍,等.肥大细胞在胃癌及癌前病变组织中的分布及其临床意义[J].川北医学院学报,2015,30(4):442-445
[12]代渊,任波,陈欢,等.肤乐颗粒对大鼠颅骨肥大细胞脱颗粒作用的拮抗效应及机制[J].中国老年学杂志,2017,37(16):3911-3914
[13]王珊,赵作涛,王誉涵,等.肥大细胞在金黄色葡萄球菌肠毒素B诱导小鼠特应性皮炎样炎症反应中的作用[J].中华皮肤科杂志,2018,51(1):26-30
[14]唐慧,王朵勤,沈燕芸,等.屋尘螨变应原对人肥大细胞脱颗粒的影响[J].上海医药,2016,37(9):10-13
[15]James C,Bernstein JA.Current and future therapies for thetreatment of histamine-induced angioedema[J].Expert Opin Pharmacother,2017,18(3):253-262
[16]席海英,霍月红.依巴斯汀联合白芍总苷、雷公藤多甙治疗慢性荨麻疹的研究[J].医学信息,2016,29(28):64-65
[17]陈文慧,刘乐,李伯埙.白芍总苷联合苯磺贝他斯汀治疗慢性荨麻疹疗效观察[J].中国中西医结合皮肤性病学杂志,2016,15(6):355-357
[18]王小艳,陈星宇,张洪英.白芍总苷在皮肤科的应用进展[J].中国中西医结合皮肤性病学杂志,2015,4(4):262-264