目的 考察艳山姜挥发油自乳化释药系统（essential oil of Fructus Alpinia zerumbet-self-emulsifying drug delivery system，EOFAZ-SEDDS）的毒性及其对人结直肠腺癌Caco-2细胞单层通透性和P-糖蛋白（P-glycoprotein，P-gp）的影响。方法 采用MTT法和乳酸脱氢酶（lactate dehydrogenase，LDH）法测定EOFAZ-SEDDS的细胞毒性；荧光黄渗透率实验考察EOFAZ-SEDDS对Caco-2单层通透性的影响；Western blotting考察EOFAZ-SEDDS对P-gp蛋白表达的影响；流式细胞仪检测EOFAZ-SEDDS对P-gp功能的影响。小鼠ig不同剂量EOFAZ-SEDDS，连续观察14 d，考察EOFAZ-SEDDS对小鼠的急性毒性作用。结果 与EOFAZ组比较，EOFAZ-SEDDS处理培养1、3、5 d的Caco-2细胞24 h后细胞存活率均显著升高（P＜0.05、0.01），LDH外漏率显著降低（P＜0.05、0.01）。与对照组比较，EOFAZ-SEDDS、单独的EOFAZ均能够显著增加荧光黄透过率（P＜0.01）；EOFAZ处理Caco-2细胞24 h后，P-gp蛋白表达显著降低（P＜0.05），EOFAZ-SEDDS处理后P-gp蛋白表达水平显著升高（P＜0.05、0.01）；10、20 μg/mL的EOFAZ-SEDDS对P-gp外排功能没有影响，40 μg/mL EOFAZ-SEDDS显著增加细胞内罗丹明的荧光强度（P＜0.05）。小鼠口服制剂的半数致死剂量（median lethal dose，LD₅₀）为5.291 5 g/kg，95%置信限为4.927 8～5.682 1 g/kg。根据急性毒性分级标准，EOFAZ-SEDDS对小鼠实际无毒。结论 EOFAZ-SEDDS可以降低EOFAZ的细胞毒性，对小鼠实际无毒，但改变了Caco-2细胞单层屏障功能，同时诱导P-gp表达，在一定浓度范围内对其功能没有影响，为EOFAZ新剂型的研究提供了一定的实验基础。

Objective To evaluate the toxicity and effects of essential oil of Fructus Alpinia zerumbet-self-emulsifying drug delivery system (EOFAZ-SEDDS) on Caco-2 cell monolayer permeability and P-glycoprotein (P-gp). Methods Cytotoxicity of EOFAZ-SEDDS on Caco-2 cell monolayers were detected by MTT and lactate dehydrogenase (LDH) methods. The effect of EOFAZ-SEDDS on Caco-2 cell monolayer permeability was investigated by fluorescence yellow permeability experiment. Western blotting was used to investigate the effect of EOFAZ-SEDDS on P-gp protein expression; The effect of EOFAZ-SEDDS on P-gp function was detected by flow cytometry. Mice were ig different doses of EOFAZ-SEDDS for 14 d, and the acute toxicity of EOFAZ-SEDDS on mice was investigated. Results Compared with EOFAZ group, survival rate of Caco-2 cells cultured for 1, 3, 5 d after EOFAZ-SEDDS treatment were significantly increased (P < 0.05, 0.01), and LDH leakage rate was significantly decreased (P < 0.05, 0.01). Compared with control group, EOFAZ-SEDDS and EOFAZ alone significantly increased the fluorescence yellow transmittance (P < 0.01). After EOFAZ treated Caco-2 cells for 24 h, P-gp protein expression was significantly decreased (P < 0.05); After EOFAZ-SEDDS treatment, P-gp protein expression was significantly increased (P < 0.05, 0.01). 10 and 20 μg/mL of EOFAZ-SEDDS had no effect on the efflux function of P-gp, while 40 μg/mL of EOFAZ-SEDDS significantly increased the fluorescence intensity of rhodamine in cells (P < 0.05). The median lethal dose (LD₅₀) of oral preparations in mice was 5.291 5 g/kg, and the 95% confidence limit was 4.927 8—5.682 1 g/kg. According to the acute toxicity grading standard, EOFAZ-SEDDS was actually non-toxic to mice. Conclusion EOFAZ-SEDDS can reduce the cytotoxicity of EOFAZ, which is actually non-toxic to mice, but it changes the monolayer barrier function of Caco-2 cells and induces the expression of P-gp, which has no effect on its function in a certain concentration range, providing a certain experimental basis for the study of new dosage forms of EOFAZ.

R285.5

贵州省科技厅自然科学计划项目（黔科合基础-ZK[2022]一般380，黔科合基础[2020]1Y210，黔科合基础-ZK[2023]一般303）；贵州医科大学国家自然科学基金培育项目（20NSP053）；贵州医科大学药学国际科技合作基地（黔科合平台人才[2017]5802）；贵州省中医药管理局中医药、民族医药科学技术研究课题（QZYY-2021-072）；贵州省卫生健康委科学技术基金项目（gzwkj2022-478）