药学学报, 2019, 54(9): 1667-1672
高嘉盼, 车德路, 张涛, 贺浪冲. 天然产物吉马酮致类过敏反应机制初步研究[J]. 药学学报, 2019, 54(9): 1667-1672.
GAO Jia-pan, CHE De-lu, ZHANG Tao, HE Lang-chong. Preliminary study on the mechanism of anaphylactoid reaction induced by natural product germacrone[J]. Acta Pharmaceutica Sinica, 2019, 54(9): 1667-1672.

高嘉盼, 车德路, 张涛, 贺浪冲
西安交通大学医学部药学院, 陕西 西安 710061
含有倍半萜类天然产物制备而成的中药注射剂在临床使用的过程中可引起多种不良反应,其中以皮肤过敏反应最为普遍。但造成这种过敏反应的原因是否与倍半萜类物质共同含有的3个异戊二烯单元相关尚不明确,因此用药安全性评价具有重要指导意义。倍半萜类物质的相对分子质量小,不是抗原或半抗原,有可能通过直接作用于肥大细胞的方式引发过敏样反应。近期研究证实,肥大细胞上的一种7次跨膜的G蛋白偶联受体(Mas-related G protein-coupled receptor-X2,MRGPRX2)是介导多种小分子药物引发过敏反应的关键靶点,不同于IgE介导的Ι型过敏反应,这种反应与给药剂量和速度具有直接相关性,首次接触致敏物即会出现过敏症状,被称为类过敏反应。本文通过一系列体外实验研究发现并不是所有倍半萜类物质均引发类过敏反应。实验选取倍半萜类物质人参皂苷Re和Rb1及吉马酮等进行钙成像筛选分析。结果表明,仅吉马酮可通过MRGPRX2激活钙动员,引起肥大细胞内钙离子浓度的增加。进一步以吉马酮为研究对象,通过β-氨基己糖苷酶释放率测定和组胺释放量测定实验证实吉马酮能够直接引起肥大细胞脱颗粒反应。进一步利用siRNA敲低MRGPRX2受体表达和吉马酮与环丙沙星的竞争性结合实验证实了作用靶点为MRGPRX2。结果表明,吉马酮可通过MRGPRX2受体直接激活肥大细胞,引发类过敏反应,这可能是含吉马酮成分中药注射剂引发皮肤过敏反应的原因。
关键词:    倍半萜类物质      吉马酮      肥大细胞      类过敏      Mas相关的G蛋白偶联受体X2     
Preliminary study on the mechanism of anaphylactoid reaction induced by natural product germacrone
GAO Jia-pan, CHE De-lu, ZHANG Tao, HE Lang-chong
School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, China
The Chinese medicine injections prepared by the natural products containing sesquiterpenoids caused various adverse reactions in clinical use, among which skin allergic reactions are the most common. However, whether the reason of allergic reaction was related to the three isoprene units contained in the sesquiterpenoids is not clear, so the evaluation of drug safety has important guiding significance. The sesquiterpenoids are small molecular substances, and they are not antigens or haptens. They may induce anaphylaxis reactions by acting mast cells directly. Current research confirmed that Mas-related G protein-coupled receptor-X2 (MRGPRX2) which is a 7-transmembrane G protein coupled receptor on mast cells was a key target mediated allergic reactions induced by many small molecular drugs. Unlike IgE-mediated allergic reactions, pseudo-allergic reaction is related to dosage and dosing rate, and occurs in the first exposure to the sensitizer. In this paper, a series of experiments in vitro found that not all sesquiterpenoids caused anaphylactoid reactions. Ginsenoside Re, ginsenoside Rb1 and germacrone were selected as representative of sesquiterpenoids for calcium imaging assay. The data confirmed that only germacrone activated calcium mobilization through MRGPRX2, causing an increase in intracellular calcium ion concentration in mast cells. Furthermore, the release rate of β-hexosaminidase and the release amount of histamine analysis confirmed that germacrone induced mast cells degranulation directly. Knockdown of MRGPRX2 expression by siRNA and competitive binding experiments against ciprofloxacin were used to prove the target of germacrone was MRGPRX2. The results indicated that germacrone could activate mast cells directly to induce anaphylactoid reaction via MRGPRX2, which might be the reason of skin allergic reactions caused by injections containing germacrone.
Key words:    sesquiterpenoid    germacrone    mast cell    pseudo-allergic reaction    Mas-related G protein-coupled receptor-X2   
收稿日期: 2019-04-23
DOI: 10.16438/j.0513-4870.2019-0323
基金项目: 国家自然科学基金重点项目(81230079).
通讯作者: 贺浪冲,Tel:86-29-82656788,E-mail:helc@mail.xjtu.edu.cn
Email: helc@mail.xjtu.edu.cn
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[1] McNeil BD, Pundir P, Meeker S, et al. Identification of a mast-cell-specific receptor crucial for pseudo-allergic drug reactions[J]. Nature, 2015, 519:237-241.
[2] Feng CX, Xiu X, Tian W, et al. Establishment of in-vitro model of complement activation and its application[J]. Acta Pharm Sin (药学学报), 2017, 52:722-728.
[3] Blanca-Lopez N, Ariza A, Doña I, et al. Hypersensitivity reactions to fluoroquinolones:analysis of the factors involved[J]. Clin Exp Allergy, 2013, 43:560-567.
[4] Xu YB, Dou DQ. Advance and prospect in studies on anaphylactoid reaction of traditional Chinese medicine injections[J]. China J Chin Mater Med (中国中药杂志), 2015, 40:2765-2773.
[5] Wang J, Wang T, Li Y S, et al. Allergic constitution theory of Chinese medicine and its assessment criterion and related studies[J]. Chin J Integr Med, 2015, 21:716-720.
[6] Hu CQ, Xu MZ, Ma Y, et al. Determination of the allergic impurities in the parenteral injection of Chinese traditional medicines containing Salvia miltiorrhiza[J]. Acta Pharm Sin (药学学报), 2008, 43:518-522.
[7] Jiang X, Lv B, Li P, et al. Bioactivity-integrated UPLC/Q-TOF-MS of Danhong injection to identify NF-κB inhibitors and anti-inflammatory targets based on endothelial cell culture and network pharmacology[J]. J Ethnopharmacol, 2015, 174:270-276.
[8] Moon P D, Choi I S, Go J H, et al. Inhibitory effects of BiRyuChe-bang on mast cell-mediated allergic reactions and inflammatory cytokines production[J]. Am J Chin Med, 2013, 41:1267-1282.
[9] Deng C, Ji J, Li N, et al. Fast determination of curcumol, curdione and germacrone in three species of Curcuma rhizomes by microwave-assisted extraction followed by headspace solid-phase microextraction and gas chromatography-mass spectrometry[J]. J Chromatogr A, 2006, 1117:115-120.
[10] Han S, Zhang T, Huang J, et al. New method of screening allergenic components from Shuanghuanglian injection:with RBL-2H3/CMC model online HPLC/MS system[J]. J Pharm Biomed Anal, 2014, 88:602-608.
[11] Braakman RB, Bezstarosti K, Sieuwerts AM, et al. Integrative analysis of genomics and proteomics data on clinical breast cancer tissue specimens extracted with acid guanidinium thiocyanate-phenol-chloroform[J]. J Proteome Res, 2015, 14:1627-1636.
[12] Ashmole I, Bradding P. Ion channels regulating mast cell biology[J]. Clin Exp Allergy, 2013, 43:491-502.
[13] Szebeni J. Complement activation-related pseudoallergy:a stress reaction in blood triggered by nanomedicines and biologicals[J]. Mol Immunol, 2014, 61:163-173.
[14] Berkes EA. Anaphylactic and anaphylactoid reactions to aspirin and other NSAIDs[J]. Clin Rev Allergy Immunol, 2003, 24:137-148.
[15] Farnam K, Chang C, Teuber S, et al. Nonallergic drug hypersensitivity reactions[J]. Int Arch Allergy Immunol, 2012, 159:327-345.
[16] Qi LW, Wang CZ, Yuan CS. American ginseng:potential structure-function relationship in cancer chemoprevention[J]. Biochem Pharmacol, 2010, 80:947-954.
[17] Sibilano R, Frossi B, Pucillo CE. Mast cell activation:a complex interplay of positive and negative signaling pathways[J]. Eur J Immunol, 2014, 44:2558-2566.