大鼠下颌骨来源的成骨细胞对普伐他汀的摄取

The transmembrane transport of pravastatin by osteoblast from rat mandible

目的:应用SD大鼠下颌骨来源的成骨细胞,观察成骨细胞对普伐他汀的摄取,研究其可能的跨膜转运机制。方法:将成年大鼠成骨细胞与溶于PBS的药物共同孵育后,弃去细胞外液,收集细胞悬液,用高效液相色谱法测定细胞内药物量,用考马斯亮蓝法测定细胞蛋白总量,以细胞内药物含量与细胞蛋白的比值作为药物转运量的观察指标。同时考察时间和药物浓度对成骨细胞摄取普伐他汀的影响,以及普伐他汀对成骨细胞OATP1(Organic anion transporting polypeptide1)转运蛋白表达的影响。结果:(1)高效相色谱法可以准确测定普伐他汀的量。(2)成骨细胞可将普伐他汀转运至细胞内。(3)成骨细胞对普伐他汀的摄取随着药物浓度及给药时间的增加呈现饱和趋势。(4)普伐他汀可上调成骨细胞OATP1的表达。结论:成骨细胞有转运普伐他汀的能力,且可能以OATP1转运载体为介导的主动转运。

Objective： To observe the transmembrane transport of pravastatin by osteoblasts from rat mandible and investigate the absorption mechanism of pravastatin. Methods： Co-culture the osteoblasts of adult rat and agents in PBS at 37℃, remove the extracellular fluid at designated times respectively, collect the suspension, then measure the intra-cellular contents of agents with high performance liquid chromatography（HPLC）, and the cell total protein with Coomassie Brilliant Blue. Uptake was given as the ratio of the cell total protein divided by the intra-cellular contents of agents. The effect of time and drug concentration on the absorption of pravastatin was studied. Results： 1, HPLC is an accurate method for the measurement of the intra-cellular contents ofpravastatin. 2, Adult rat osteoblasts has the ability of intra-cellular transport of pravastatin.3, The absorption of pravastatin could be saturable with time and drug concentration increasing.4, OATP1 was expressed in osteoblasts and the protein level of OATP1 can be up-regulated by pravastatin. Conclusions： Pravastatin can be transported into osteoblasts from SD rat＇s mandible, and the absorption ofpravastatin in osteoblasts may be mediated by OATP1.