体外研究黄酮类和芪类化合物对血脑屏障P-糖蛋白外排沙奎那韦的影响

Effects of several flavonoids and stilbenes on P-glycoprotein mediated efflux of saquinavir on blood-brain barrier in vitro

目的研究9种黄酮类化合物和5种芪类化合物对血脑屏障上P-糖蛋白(P-gp)外排沙奎那韦的影响。方法以过表达P-gp的马丁达比犬肾上皮细胞(MDCKII-MDR1)作为血脑屏障体外模型。药物干预分为16组(n=4),包括阴性对照组、阳性对照组、黄酮类实验I-IX组和芪类实验组Ⅰ-Ⅴ组。药物干预分别为50μmol·L^(-1)沙奎那韦(阴性对照组);50μmol·L^(-1)沙奎那韦联用50μmol·L^(-1)维拉帕米(阳性对照组);50μmol·L^(-1)沙奎那韦联用25μmol·L^(-1)黄酮类(黄酮类实验I-IX组)或联用25μmol·L^(-1)芪类化合物(芪类实验组Ⅰ-Ⅴ组)。药物干预时间均为4 h。用液相色谱质谱联用技术测定沙奎那韦在各组细胞内的积聚量。结果细胞内沙奎那韦积聚量:与阴性对照组(101.46±22.00)ng·mg^(-1)相比,柚皮素组(229.41±74.86)ng·mg^(-1)、桑黄素组(149.89±30.38)ng·mg^(-1)、白杨黄素组(209.75±65.35)ng·mg^(-1)、氧化白藜芦醇组(178.54±44.40)ng·mg^(-1)、染料木黄酮组(294.49±66.35)ng·mg^(-1)、表儿茶素组(212.27±49.70)ng·mg^(-1)、三乙酰白藜芦醇组(290.94±45.54)ng·mg^(-1)和白藜芦醇三甲醚组(205.55±35.16)ng·mg^(-1),均显著升高,差异有统计学意义(P<0.05);异槲皮苷组(61.98±11.02)ng·mg^(-1)、槲皮素组(41.54±4.92)ng·mg^(-1)和白藜芦醇组(29.59±4.61)ng·mg^(-1)细胞内沙奎那韦积聚量,均显著降低,差异有统计学意义(P<0.05)。结论一些黄酮类、芪类化合物可能对P-gp介导的沙奎那韦在血脑屏障上的转运过程产生调节作用。

Objective To investigate the effects of 9 flavonoids and 5 stilbenes on the P - glycoprotein （ P - gp） mediated efflux of saquinavir on blood - brain barrier （BBB） in vitro. Methods Madin - Darby ca- nine kidney cells （MDCKII -MDR1 ） expressing P-gp were performed as an in vitro blood - brain barrier （ BBB ） model. Incubations were divi- ded into 16 groups （n =4）, including a negative control group, a posi- tive control group, 9 flavonoid intervention groups, and 5 stilbene inter- vention groups. In negative control group, cells were incubated in 50 μmol · L^-1 saquinavir. In positive control group, cells were incubated in 50 μmol · L^-l saquinavir in the presence of 50 μmol ·L^-1 verapamil. In flavonoid intervention groups （ I - IX ）, cells were incubated in 50 μmol · L^-1 saquinavir in the presence of 25 μmol ·L^-lcertain flavonoid.In stilbene intervention groups （ I - V ）, cells were incubated in 50 μmol · L^-1 saquinavir in the presence of 25 μmol · L^-1 certain stilbene. Incubations were terminated after 4 h. The intracellular concentrations of saquinavir were determined using a high - performance liquid chromatography - tandem mass spectrometry （ HPLC - MS/MS） method. Results Compared with the negative control group （101.46 ± 22. 00） ng · mg^-1, the accmnulation of saquinavir in MDCKII - MDRI cells in the presence of naringenin （229. 41± 74. 86 ） ng ·mg^-1, morinhydrate （ 149. 89 ± 30. 38 ） ng·mg^-1 chrysin （209.75 ± 65. 35） ng · rng^-1 , genistein ng.mg , , and oxyresveratrol （178.54 ± 44.40） ng ·mg^-1 （294.49±66.35） ng · mg^-1, L - epicatechin （212.27 ±49.70） ng · mg-^1, triacetyl - resveratrol （290.94 ±45.54） -1 1 ng· mg , and 3,4＇ ,5 - trimethoxy - trans - stilbene （205. 55 ±35. 16） ng · mg- increased significantly （P 〈 0. 05 ）. In contrast, the accumulation of saquinavir in MDCKII -MDR1 cells in the presence ofisoquercitrin （61.98 ± 11.02 ） ng· mg^-1 , quercetin （41.54±4.92） ng · rng^-1 , and resveratrol （29.59 ±4.61） ng · mg^-1 decreased significantly （P〈0.05）. Conclusion Several flavonoids and stilbenes may exert modulaory efiects on P-gp mediated efflux of saquinavir on BBB.