目的 為了提高木芙蓉Hibiscus mutabilis葉的臨床療效，對其有效組分進行配伍研究并建立設計空間，為制劑成型工藝研究奠定基礎。方法 體外培養(yǎng)牛乳腺上皮細胞（ BMEC），以CCK-8法檢測的細胞存活率和試劑盒法檢測的腫瘤壞死因子（ TNF）-α、白細胞介素（ IL）-1β分泌量為指標，篩選脂多糖（ LPS）造模的適宜濃度及時間；應用建立的BMEC細胞的LPS模型，以細胞存活率和TNF-α、IL-1β分泌量為指標，篩選出木芙蓉葉抗急性乳腺炎有效組分——蘆丁、異槲皮苷、槲皮素、山柰酚、金絲桃苷的最佳配伍組合；以細胞存活率為指標，采用單因素實驗初步篩選最佳組合成分中蘆丁、異槲皮苷、山柰酚抗LPS誘導BMEC細胞的有效給藥濃度；通過Box-Behnken設計建立模型，采用多指標疊加法建立蘆丁、異槲皮苷、山柰酚抗LPS誘導BMEC細胞的設計空間，并以細胞存活率和細胞上清中TNF-α、IL-1β、IL-6含量為指標，對隨機的選取空間點內和點外進行驗證，篩選出給藥濃度。結果 10 μg·mL^-1 LPS刺激BMEC細胞24 h制備模型；木芙蓉葉抗急性乳腺炎有效組分的最佳配伍組合為蘆丁、異槲皮苷和山柰酚3個成分；建立的設計空間蘆丁的質量濃度為96～104 μg·mL^-1、異槲皮苷的質量濃度為45～54 μg·mL^-1、山柰酚的質量濃度為4.5～6.5 μg·mL^-1，經(jīng)驗證，在此空間內的點均在所設的預測范圍內。結論 成功篩選出木芙蓉葉有效組分的最佳配伍，且建立的設計空間工藝穩(wěn)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.

R285.5

貴州省基礎研究計劃（自然科學類）（黔科合基礎-ZK［2023］一般407）；貴州中醫(yī)藥大學外用制劑研究中心；貴州省中醫(yī)藥管理局中醫(yī)藥、民族醫(yī)藥科學技術研究專項課題（QYZZ-2025-066）；畢節(jié)市科技計劃項目（畢科合（2024）15號）