目的 为了提高木芙蓉Hibiscus mutabilis叶的临床疗效，对其有效组分进行配伍研究并建立设计空间，为制剂成型工艺研究奠定基础。方法 体外培养牛乳腺上皮细胞（ BMEC），以CCK-8法检测的细胞存活率和试剂盒法检测的肿瘤坏死因子（ TNF）-α、白细胞介素（ IL）-1β分泌量为指标，筛选脂多糖（ LPS）造模的适宜浓度及时间；应用建立的BMEC细胞的LPS模型，以细胞存活率和TNF-α、IL-1β分泌量为指标，筛选出木芙蓉叶抗急性乳腺炎有效组分——芦丁、异槲皮苷、槲皮素、山柰酚、金丝桃苷的最佳配伍组合；以细胞存活率为指标，采用单因素实验初步筛选最佳组合成分中芦丁、异槲皮苷、山柰酚抗LPS诱导BMEC细胞的有效给药浓度；通过Box-Behnken设计建立模型，采用多指标叠加法建立芦丁、异槲皮苷、山柰酚抗LPS诱导BMEC细胞的设计空间，并以细胞存活率和细胞上清中TNF-α、IL-1β、IL-6含量为指标，对随机的选取空间点内和点外进行验证，筛选出给药浓度。结果 10 μg·mL^-1 LPS刺激BMEC细胞24 h制备模型；木芙蓉叶抗急性乳腺炎有效组分的最佳配伍组合为芦丁、异槲皮苷和山柰酚3个成分；建立的设计空间芦丁的质量浓度为96～104 μg·mL^-1、异槲皮苷的质量浓度为45～54 μg·mL^-1、山柰酚的质量浓度为4.5～6.5 μg·mL^-1，经验证，在此空间内的点均在所设的预测范围内。结论 成功筛选出木芙蓉叶有效组分的最佳配伍，且建立的设计空间工艺稳定可靠。

Objective To improve the clinical efficacy of H. manihot leaves, the effective components were studied for compatibility and a design space was established to lay the foundation for subsequent formulation and process research. Methods Bovine mammary epithelial cells (BMEC) were cultured in vitro. The appropriate concentration and time of lipopolysaccharide (LPS) for model establishment were screened by cell viability detected by CCK-8 and the secretion of TNF-α and IL-1β detected by kits. Using the established LPS model of BMEC cells, the best compatibility combination of effective components of H. manihot leaves against acute mastitis (rutin, isorhamnetin, quercetin, kaempferol and hyperoside) was screened by cell viability and the secretion of TNF-α and IL- 1β. The effective drug concentration of rutin, isorhamnetin and kaempferol against LPS-induced BMEC cells was preliminarily screened by single factor experiments with cell viability as the index. A model was established by Box-Behnken design, and the design space of rutin, isorhamnetin and kaempferol against LPS-induced BMEC cells was established by multi-index superposition method. The random points within and outside the space were verified by cell viability and the content of TNF-α, IL-1β and IL-6 in the cell supernatant as the index, and the drug concentration was screened. Results The model was established by stimulating BMEC cells with 10 μg·mL^-1 LPS for 24 h; the best compatibility combination of effective components of H. manihot leaves against acute mastitis was rutin, isorhamnetin and kaempferol; the established design space was rutin mass concentration of 96—104 μg·mL^-1, isorhamnetin mass concentration of 45—54 μg·mL^-1, and kaempferol mass concentration of 4.5—6.5 μg·mL^-1. All the points within this space were within the predicted range after verification. Conclusion The best compatibility of effective components of H. manihot leaves was successfully screened, and the established design space process was stable and reliable.

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贵州省基础研究计划（自然科学类）（黔科合基础-ZK［2023］一般407）；贵州中医药大学外用制剂研究中心；贵州省中医药管理局中医药、民族医药科学技术研究专项课题（QYZZ-2025-066）；毕节市科技计划项目（毕科合（2024）15号）