体外抗过敏作用研究方法优化及中药药对抗过敏活性筛选  

Optimization of Methods for Study of Antiallergic Activity in vitro and Screening of Chinese Medicine Anti-Allergic Activity

作　　者：李依林 左泽平 田颖颖 刘闯 赵新月 吕英楠 王志斌[1,2] LI Yilin;ZUO Zeping;TIAN Yingying;LIU Chuang;ZHAO Xinyue;LYU Yingnan;WANG Zhibin(School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102401,China;Beijing Zhongyan Tong Ren Tang Technology Development Co.,LTD.,Beijing 100071,China)

机构地区：[1]北京中医药大学中药学院,北京102401 [2]北京中研同仁堂科技发展股份有限公司,北京100071

出　　处：《世界科学技术-中医药现代化》2024年第5期1261-1268,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家科学技术部国家重点研发计划(2019YFC1711400):经典名方标准颗粒制备与标准研究,负责人:刘晖晖;国家科学技术部国家科技重大专项(民口)课题(2018ZX09201-011):中药先进制药与信息化技术融合示范研究,负责人:李正

摘　　要：目的优化两种中药体外抗过敏活性筛选方法,比较5个中药药对的体外抗过敏活性。方法优化RBL-2H3(Rat basophil leukemia cells)细胞脱颗粒实验方法,通过甲苯胺蓝染色以及β-氨基己糖苷酶(β-hexosaminidase,β-HEX)和组胺(Histamine,HIS)释放3个指标比较5个药对抑制肥大细胞脱颗粒的活性;优化了透明质酸酶抑制实验,对5个中药药对的透明质酸酶抑制作用进行比较。结果在RBL-2H3细胞脱颗粒实验中,5个中药药对中荆芥-防风、麻黄-细辛、防风-白芷、川芎-细辛均能不同程度抑制模型细胞释放β-HEX(P<0.05),苍耳子-辛夷与细胞共培养后脱颗粒模型细胞上清中β-HEX释放率显著升高(P<0.05);5个中药药对均可以降低脱颗粒模型细胞上清中HIS释放量。透明质酸酶抑制率实验中,各药对的透明质酸酶抑制率川芎-细辛>防风-白芷>荆芥-防风>麻黄-细辛,其中川芎-细辛抗过敏活性最强。结论两种方法获得的不同药对体外抗过敏活性趋势具有一致性,说明两种方法可以用于体外抗过敏活性的筛选。Objective To optimize the screening methods for antiallergic activity of two Chinese medicines in vitro and compare the antiallergic activity of five medicine pairs in vitro.Methods The degranulation assay of RBL-2H3 cells was optimized,and the activity of the five drugs on inhibiting mast cell degranulation was compared by toluidine blue staining and the release ofβ-HEX and histamine(HIS).The hyaluronidase inhibition test was optimized to compare the hyaluronidase inhibition effect of five Chinese medicine pairs.Results In the experiment of degranulation of RBL-2H3 cells,5 medicine pairs could inhibitβ-HEX release from model cells to different degrees(P<0.05),including Schizonepetae Herba-Saposhnicovia divaricata,Ephedrae Herba-Asarum,Saposhnicovia divaricata-Radix Angelicae dahuricae,Rhizoma Chuanxiong-Asarum.Theβ-HEX release rate in the supernatant of degranulated model cells was significantly increased(P<0.05)after coculture with Flos Magnoliae-Fructus Xanthii.All the five medicine pairs could reduce HIS release in the supernatant of degranulated model cells.In the hyaluronase inhibition rate test,the hyaluronase inhibition rate of each medicine pair was Rhizoma Chuanxion-Asarum>Saposhnicovia divaricate-Radix Angelicae dahuricae>Schizonepetae Herba-Saposhnicovia divaricate>Ephedrae Herba-Asarum,among which Rhizoma Chuanxiong-Asarum had the strongest anti-allergic activity.Conclusion The trend of antiallergic activity of different drugs obtained by the two methods is consistent in vitro,indicating that the two methods can be used for screening antiallergic activity in vitro.

关 键 词：抗过敏 透明质酸酶抑制 肥大细胞脱颗粒实验 RBL-2H3细胞 

分 类 号：R285.5[医药卫生—中药学]