香丹注射液中致类过敏反应成分筛选及机制初探  

Screening and preliminary mechanism of inducer of anaphylactic reaction in Xiangdan Injection

作　　者：庞菲 王弯弯 郝瑞瑞 周甜雨 陈世忠[2] 靳洪涛[1,4,5] PANG Fei;WANG Wan-wan;HAO Rui-rui;ZHOU Tian-yu;CHEN Shi-zhong;JING Hong-tao(New Drug Safety Evaluation Center,Institute of Materia Medica,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050,China;School of Pharmaceutical Sciences,Peking University,Beijing 100191,China;Shaanxi University of Chinese Medicine,Xianyang 712046,China;NMPA Key Laboratory for Safety Research and Evaluation of Innovative Drug,Beijing 102206,China;Beijing Union-Genius Pharmaceutical Technology Development Co.,Ltd.,Beijing 100176,China)

机构地区：[1]中国医学科学院,北京协和医学院药物研究所,新药安全评价研究中心,北京100050 [2]北京大学药学院,北京100191 [3]陕西中医药大学,陕西咸阳712046 [4]NMPA创新药物安全性研究与评价重点实验室,北京102206 [5]北京协和建昊医药技术开发有限责任公司,北京100176

出　　处：《中草药》2022年第20期6500-6508,共9页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81773996);中国毒理学会临床毒理专项资助课题(CST2021CT101)。

摘　　要：目的 筛选香丹注射液中可能存在的致类过敏成分并探讨其作用机制。方法 使用Auto Dock Tools Vina软件,以人类Mas相关G蛋白偶联受体X2(Mas-related G protein-coupled receptor X2,MRGPRX2)和小鼠MRGPRb2为目标蛋白,对香丹注射液中的水溶性酚酸类物质进行分子对接,并将香丹注射液及与MRGPRX2和MRGPRb2有较高结合(≤-25.10k J/mol)的组成成分丹酚酸A、丹酚酸B、丹酚酸C、迷迭香酸、紫草酸与小鼠肥大细胞瘤P815细胞孵育1 h,检测组胺(histamine,His)和β-氨基己糖苷酶(β-hexosaminidase,β-Hex)释放率,筛选出致类过敏成分,进一步观察丹酚酸B及紫草酸刺激P815细胞胞内Ca^(2+)动员及细胞脱颗粒情况,最后进行小鼠后爪外渗实验验证。结果 P815细胞分别给予香丹注射液(1:25、1:250)及丹酚酸B(100μg/mL)和紫草酸(100μg/mL)刺激后,His、β-Hex释放均显著升高(P<0.01)。P815细胞分别给予丹酚酸B(100μg/mL)和紫草酸(100μg/mL)刺激后,胞内Ca^(2+)浓度均显著升高(P<0.05),并有脱颗粒现象。丹酚酸B(43.5、21.7μg/kg)和紫草酸(43.5μg/kg)均能显著提高小鼠后爪伊文思蓝渗出程度(P<0.05、0.01)。结论 香丹注射液诱导的类过敏反应可能与丹酚酸B和紫草酸有关,且丹酚酸B和紫草酸诱导的类过敏反应可能与动员Ca^(2+)内流相关。Objective To screen the components induced the anaphylactic reaction in Xiangdan Injection(香丹注射液)and explore the mechanism.Methods With human Mas-related G protein-coupled receptor X2(MRGPRX2)and mouse MRGPRb2 as target proteins,water-soluble phenolic acids in Xiangdan Injection were docked with target protein by Auto Dock Tools Vina.P815 cells were incubated with Xiangdan Injection and its constituents of salvianolic acid A,salvianolic acid B,salvianolic acid C,rosmarinic acid and lithospermic acid for 1 h,which had high binding energy(≤−25.10 kJ/mol)to MRGPRX2 and MRGPRb2.According tohistamine (His) and β-hexosaminidase (β-Hex) release rate,possible components that may induce the anaphylactic reaction werescreened out. The intracellular Ca^(2+) mobilization and cell degranulation stimulated by salvianolic acid B and lithospermic acid werefurther observed. Finally,the mouse hind paw extravasation experiment was observed for verification. Results After P815 cells werestimulated with Xiangdan Injection (1∶25,1∶250),salvianolic acid B (100 μg/mL) and lithospermic acid (100 μg/mL),the releaseof His and β-Hex were significantly increased (P < 0.01). After P815 cells were stimulated with salvianolic acid B (100 μg/mL) andlithospermic acid (100 μg/mL),the intracellular Ca^(2+) concentration was significantly increased (P < 0.05),and degranulation wasobserved. Salvianolic acid B (43.5,21.7 μg/kg) and lithospermic acid (43.5 μg/kg) could significantly increase the degree of Evansblue exudation in hind paw of mice (P < 0.05,0.01). Conclusion The anaphylactoid reaction induced by Xiangdan Injection may berelated to salvianolic acid B and lithospermic acid,and the anaphylactoid reaction induced by salvianolic acid B and shikonic acid maybe related to the mobilization of Ca^(2+) influx.

关 键 词：香丹注射液 类过敏反应 分子对接 丹酚酸B 紫草酸 P815细胞 

分 类 号：R285.5[医药卫生—中药学]