目的 制备花旗松素单体并测定花旗松素在不同介质中的表观油水分配系数，研究其在结肠癌Caco-2细胞模型中吸收转运机制。方法 酶解法制备花旗松素，采用高效液相色谱法测定花旗松素在37℃、不同pH值缓冲盐溶液和水中的饱和溶解度，通过油水分配系数计算公式得花旗松素表观油水分配系数；CCK-8实验考察花旗松素在Caco-2细胞中的安全浓度范围，再利用Caco-2细胞单层模型研究其双侧跨膜吸收转运机制；CCK-8实验考察花旗松素在人脐血管内皮细胞HDMEC细胞中的安全浓度范围，采用脂多糖诱导的HDMEC细胞建立炎症模型，用花旗松素进行干预，乳酸脱氢酶（LDH）试剂盒检测LDH的活力。结果 花旗松素在不同溶剂中的lgP分别为0.29（0.1 mol/L盐酸）、0.48（pH 2.0）、0.46（pH 5.8）、0.34（pH 6.8）、0.26（pH 7.4）、0.38（水）；花旗松素质量浓度在50～500 μg/mL对Caco-2细胞无明显毒性作用；不同质量浓度花旗松素在Caco-2单层细胞模型中的转运具有时间依赖性，不同质量浓度花旗松素双侧转运P_app值无显著性差异，且均小于1×10^-6 cm/s，外排率（ER）均小于2。花旗松素在50～300 μg/mL对HDMEC细胞无明显毒性作用，给予花旗松素干预后，与单独给予脂多糖（LPS）刺激组相比，花旗松素各用药组均能明显降低LDH活力（P<0.05），花旗松素在50～100 μg/mL明显降低LDH活力，100～250 μg/mL趋于平稳。结论 花旗松素在肠道中属于难吸收药物，其跨膜转运机制为被动转运，其有对LPS致HDMEC细胞炎症有抑制作用，具有抗炎活性。

Objective To prepare the taxifolin and determine its apparent oil-water partition coefficient in different media, and to study the mechanism of absorption and transport of taxifolin in Caco-2 cell model. Methods Taxifolin was prepared by enzymolysis. HPLC was used to determine the saturated solubility of taxifolin in 37℃, different pH buffer solution and water, apparent oil-water distribution coefficient of taxifolin obtained by calculation formula of oil-water distribution coefficient; CCK-8 experiment was used to investigate the safe concentration range of taxifolin in Caco-2 cells, and then the single-layer model of Caco-2 cells was used to study the mechanism of bilateral transmembrane absorption and transport. CCK-8 experiment was used to investigate the safe concentration range of taxifolin in HDMEC cells. The inflammatory model of HDMEC cells induced by lipopolysaccharide was established, and the activity of lactic dehydrogenase was detected by the intervention of floxacin. The activity of lactic dehydrogenase was detected by lactic dehydrogenase kit. Results The lgP values of taxifolin in the following solvents were 0.29 (0.1 mol/L hydrochloric acid), 0.48 (pH 2.0), 0.46 (pH 5.8), 0.34 (pH 6.8), 0.26 (pH 7.4), and 0.38 (water), respectively; There was no significant toxic effect on Caco-2 cells in the range of 50-500 μg/mL; There was no significant difference in P_app value of bilateral transport between different concentrations of taxifolin in Caco-2 monolayer cell model, and it was less than 1×10^-6 cm/s and ER was less than 2. There was no significant toxic effect on HDMEC cells in the range of 50-300 μg/mL; After treatment with taxifolin, compared with LPS stimulation group, the activity of LDH in each treatment group was decreased significantly (P<0.05), and the activity of LDH was decreased significantly in the range of 50-100 μg/mL, and tended to be stable in the range of 100-250 μg/mL. Conclusion Taxifolin is a kind of drug which is difficult to absorb in the intestine. The mechanism of transmembrane transport is passive transport. It can inhibit the inflammation of hdmec cells induced by LPS and has anti-inflammatory activity.

R285.5

国家新药创新项目（2017ZX09301062）