目的 研究丹红注射液中主要成分羟基红花黄色素A（HSYA）在Caco-2细胞单层模型的转运特征。方法 MTT法确定HSYA对Caco-2细胞单层模型作用的安全浓度范围；采用Caco-2细胞单层模型考察转运时间、药物质量浓度、温度、pH值以及P-糖蛋白（P-gp）抑制剂维拉帕米和能量代谢抑制剂叠氮化钠对HSYA转运的影响；RT-PCR法检测HSYA及维拉帕米对多药耐药基因（MDR1）表达的影响。结果 从顶侧（AP）到底侧（BL）（AP→BL），HSYA的表观渗透系数（P_app）在2×10^-6～5×10^{-6 cm/s}，表明其吸收性中等；HSYA的转运与其质量浓度和时间呈正相关，37℃下HSYA的P_app与4、25℃下的P_app有显著差异（P<0.01），pH值为9.0时的P_app与pH值为5.0、7.4时的P_app值也有明显差异（P<0.01）。维拉帕米能明显下调MDR1基因的表达，但HSYA的转运不受维拉帕米的影响；叠氮化钠影响细胞能量代谢，但HSYA的转运不受能量代谢异常的影响，且P_app(BL→AP)/P_app(AP→BL)在1～1.5，HSYA的吸收过程基本符合被动扩散。结论 HSYA在Caco-2细胞模型的转运方式为被动扩散，且不受P-gp和能量代谢的影响，低温和碱性环境下不利于HSYA的吸收。

Objective To investigate the transport characteristics of hydroxysafflor yellow A (HSYA) in Danhong Injection across Caco-2 cell monolayer. Methods Safe concentration range of HSYA against Caco-2 cell monolayer model was selected by MTT method; The effects of time, drug concentration, temperature, pH, P-gp inhibitor (Verapamil), and energy metabolism inhibitor (sodium azide) on the absorption of HSYA were observed by Caco-2 cell monolayer model; The multidrug resistance (MDR1) gene expression in Caco-2 cells was analyzed by the RT-PCR method. Results The P_app of HSYA transport from apical (AP) side to basolateral (BL) side was in 2×10^-6 -5×10^{-6 cm/s, which showed a medium absorption. The transport of HSYA was positively correlated with time and concentration. The P}_app of HSYA transport at 37℃ has significant differences with those at 4 and 25℃ (P < 0.01). The P_app of HSYA transport under pH 9.0 has significant differences with those under pH 5.0 and 7.4 (P < 0.01). The gene expression of MDR1 was significantly reduced by Verapamil, but the transport of HSYA was not influenced by Verapamil and sodium azide, the number of P_app(BL→AP)/P_app(AP→BL) was between 1 and 1.5, so the absorption of HSYA was basically in line with the passive diffusion. Conclusion The transport of HSYA across Caco-2 cell monolayer model is passive diffusion, and is not influenced by the change of P-gp and energy metabolism. Low temperature and alkaline environment are not conducive to the absorption of HSYA.

国家自然科学基金资助项目（81373898，81173647，81202636，81274176）；浙江省自然科学基金资助项目（LR12H27001）；浙江省科技厅项目（2013C33244）；浙江省卫生高层次创新人才培养工程项目；浙江省中医药（中西医结合）重点学科项目（2012-XK-A06）；浙江省教育厅科研资助项目（Y201329166）