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（≤−25.10 kJ/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Ca²⁺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（100 μg/mL）和紫草酸（100 μg/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（100 μg/mL）和紫草酸（100 μg/mL）刺激后，胞内Ca²⁺浓度均显著升高（P＜0.05），并有脱颗粒现象。丹酚酸B（43.5、21.7 μg/kg）和紫草酸（43.5 μg/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Ca²⁺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 to histamine (His) and β-hexosaminidase (β-Hex) release rate, possible components that may induce the anaphylactic reaction were screened out. The intracellular Ca²⁺ mobilization and cell degranulation stimulated by salvianolic acid B and lithospermic acid were further observed. Finally, the mouse hind paw extravasation experiment was observed for verification. Results After P815 cells were stimulated with Xiangdan Injection (1∶25, 1∶250), salvianolic acid B (100 μg/mL) and lithospermic acid (100 μg/mL), the release of His and β-Hex were significantly increased (P < 0.01). After P815 cells were stimulated with salvianolic acid B (100 μg/mL) and lithospermic acid (100 μg/mL), the intracellular Ca²⁺ concentration was significantly increased (P < 0.05), and degranulation was observed. Salvianolic acid B (43.5, 21.7 μg/kg) and lithospermic acid (43.5 μg/kg) could significantly increase the degree of Evans blue exudation in hind paw of mice (P < 0.05, 0.01). Conclusion The anaphylactoid reaction induced by Xiangdan Injection may be related to salvianolic acid B and lithospermic acid, and the anaphylactoid reaction induced by salvianolic acid B and shikonic acid may be related to the mobilization of Ca²⁺ influx.

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)