DataSheet7_Compound Kushen Injection Induces Immediate Hypersensitivity Reaction Through Promoting the Production of Platelet-Activating Factor via de Novo Pathway.XLSX

Compound Kushen Injection (CKI) is a bis-herbal formulation extracted from Kushen (Radix Sophorae Flavescentis) and Baituling (Rhizoma Heterosmilacis Yunnanensis). Clinically, it is used as the adjuvant treatment of cancer. However, with the increased application, the cases of immediate hypersensitivity reactions (IHRs) also gradually rise. In this study, we investigated the underlying mechanism(s) and active constituent(s) for CKI-induced IHRs in experimental models. The obtained results showed that CKI did not elevate serum total IgE (tIgE) and mouse mast cell protease 1 (MMCP1) after consecutive immunization for 5 weeks, but could induce Evans blue extravasation (local) and cause obvious hypothermia (systemic) after a single injection. Further study showed that alkaloids in Kushen, especially matrine, were responsible for CKI-induced IHRs. Mechanism study showed that various platelet-activating factor (PAF) receptor antagonists could significantly counter CKI-induced IHRs locally or systemically. In cell system, CKI was able to promote PAF production in a non-cell-selective manner. In cell lysate, the effect of CKI on PAF production became stronger and could be abolished by blocking de novo pathway. In conclusion, our study identifies, for the first time, that CKI is a PAF inducer. It causes non-immunologic IHRs, rather than IgE-dependent IHRs, by promoting PAF production through de novo pathway. Alkaloids in Kushen, especially matrine, are the prime culprits for IHRs. Our findings may provide a potential approach for preventing and treating CKI-induced IHRs.

复方苦参注射液（Compound Kushen Injection，CKI）是由苦参（Radix Sophorae Flavescentis）与白土苓（Rhizoma Heterosmilacis Yunnanensis）提取得到的双药复方制剂。临床中，其被用作癌症的辅助治疗手段。然而随着应用范围扩大，速发型超敏反应（immediate hypersensitivity reactions, IHRs）的报道病例数也逐渐增多。本研究通过实验模型，探究了CKI诱发速发型超敏反应的潜在机制与活性成分。研究结果显示，连续免疫5周后，CKI并未升高小鼠血清总免疫球蛋白E（total IgE, tIgE）与小鼠肥大细胞蛋白酶1（mouse mast cell protease 1, MMCP1）水平，但单次给药后可诱导局部伊文思蓝渗出，并引发明显的全身性体温过低。进一步研究表明，苦参中的生物碱类成分，尤其是苦参碱，是CKI诱发速发型超敏反应的关键物质。机制探究显示，多种血小板活化因子（platelet-activating factor, PAF）受体拮抗剂可显著拮抗CKI诱发的局部或全身性速发型超敏反应。在细胞体系中，CKI能够以非细胞选择性的方式促进PAF的生成。在细胞裂解液中，CKI对PAF生成的促进作用更为显著，且该效应可通过阻断从头合成途径（de novo pathway）被消除。综上，本研究首次证实CKI是一种PAF诱导剂。其通过从头合成途径促进PAF生成，从而引发非免疫依赖性的速发型超敏反应，而非IgE依赖型速发型超敏反应。苦参中的生物碱类成分，尤其是苦参碱，是诱发该类超敏反应的主要元凶。本研究结果可为CKI诱发的速发型超敏反应的防治提供潜在策略。