盐酸氯丙嗪对caco-2细胞膜流动性的影响-第三军医大学学报.doc

PAGE PAGE 2 作者简介：杨丹，女，硕士研究生，药剂学专业 *通讯作者：王章阳，男，博士，教授，主要从事药物新剂型的研究 Tel：（023 *Email：cgwangzy@163.com 盐酸氯丙嗪促进药物直肠吸收机制研究 杨丹，王章阳*，陈勇川，陈剑鸿，戴青，刘松青，夏培元（第三军医大学西南医院药学部 重庆 400038） 摘要：目的　前期研究结果表明盐酸氯丙嗪能够促进药物在直肠的吸收。为进一步了解其作用机理，应用Caco-2细胞（the human colon adenocarcinoma cell lines）考察盐酸氯丙嗪对直肠粘膜上皮细胞完整性与稳定性的影响。方法 以不同浓度的盐酸氯丙嗪作用Caco-2细胞后，采用Bradford法测定上清液OD值，计算细胞膜破坏百分率以了解Caco-2细胞膜完整性的变化；应用DPH（1,6-diphenyl-1,3,5-hexatriene）荧光探针法测定细胞膜脂质DPH的荧光强度并计算荧光偏振度，评价细胞膜流动性的变化以了解其对Caco-2细胞膜稳定性的影响。结果 盐酸氯丙嗪浓度＞140μmol·L-1时能影响Caco-2细胞膜完整结构，使细胞内容物释出，且浓度与对膜破坏程度呈现S型曲线。与对照组比较，盐酸氯丙嗪＜112μmol·L-1时造成Caco-2细胞膜脂质荧光偏振度值降低，并随浓度的增高及作用时间的延长，呈逐渐下降的趋势（P0.05）。结论 盐酸氯丙嗪在低浓度时能够增大Caco-2细胞膜的流动性，使其结构紊乱，影响膜的稳定性；当超过一定浓度时可破坏细胞膜的完整性，使细胞内容物释出；这些改变很可能是盐酸氯丙嗪促进药物跨膜转运吸收的机理之一。 关键词 盐酸氯丙嗪；直肠吸收；细胞膜；机理； A study on mechanism of Chlorpromazine Hydrochloride promotes the absorption of drugs in rectum YANG Dan,WANG Zhang-yang*,CHEN Yong-chuan,CHEN Jian-hong,DAI Qing,LIU Song-qing,XIA Pei-yuan（Department of Pharmacy , The Affiliated Southwest Hospital of Third Military Medical University, Chongqing 400038 , China） Abstract：Objective Our previous research have shown that Chlorpromazine Hydrochloride can increase the relative bioavailability of drugs by rectal administration, and this alternation could be a consequence of altered cell membrane architecture and permeability caused by the amphipathic structure of Chlorpromazine Hydrochloride.To probe this,the effect of Chlorpromazine Hydrochloride on Caco-2 cell membrane was investigated in the present study. Methods After the Caco-2 cells were treated with different concentrations of Chlorpromazine Hydrochloride at different durations, the OD-value of the supernatant was determinated by Bradford potein assays, then calculated the OD ratio to display the changes on Caco-2 cell membrane integrity.And the fluorescent intensity was determinated by fluorescence spectrophotometer with 1,6-diphenyl-1,3,5-hexatriene(DPH) as a fluorescein mark, thus to calculate the fluorescence polariz