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目的应用Caco-2细胞(the human colon adenocarcinoma cell lines)考察盐酸氯丙嗪对直肠黏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法测定上清液吸光度值[D(595)],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。盐酸氯丙嗪浓度<112μmol/L时Caco-2细胞膜脂质荧光偏振度值随浓度的增高及作用时间的延长,呈逐渐下降的趋势(P<0.05)。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制之一。
Objective To investigate the effect of chlorpromazine hydrochloride on the integrity and stability of rectal mucosal epithelial cells by Caco-2 cells (the human colon adenocarcinoma cell lines). Methods After the Caco-2 cells were treated with different concentrations of chlorpromazine hydrochloride, the absorbance of the supernatant was measured by Bradford method [D (595)], and the percentage of cell membrane destruction was calculated to understand the membrane integrity of Caco-2 cells. 1,6-diphenyl-1,3,5-hexatriene) fluorescence probe method to measure the fluorescence intensity of the cell membrane lipid DPH and calculate the fluorescence polarization degree, evaluate the change of cell membrane fluidity to understand its stability to Caco-2 cell membrane influences. Results When the concentration of chlorpromazine hydrochloride was> 140μmol / L, the integrity of Caco-2 cell membrane was affected and the cell contents were released. The concentration and the degree of damage to membrane were S curve. When the concentration of chlorpromazine hydrochloride was lower than 112μmol / L, the polarization degree of lipid in Caco-2 cells decreased gradually with increasing concentration and prolonging of acting time (P <0.05). Conclusion Chlorpromazine hydrochloride can increase the fluidity of Caco-2 cell membrane at low concentration, and cause disorder of its structure, affecting the stability of membrane. When the concentration of chlorpromazine exceeds a certain concentration, the integrity of cell membrane can be destroyed and cell contents can be released. These changes are likely to be one of the mechanisms by which chlorpromazine hydrochloride promotes transmembrane transport of drugs.