摘要
目的：观察黄芪多糖（APS）对Caco-2细胞P糖蛋白（P-gp）功能、表达的影响。方法采用MTT法考察药物对细胞的毒性,确定药物最大无毒浓度；流式细胞仪测定Caco-2细胞P-gp底物罗丹明-123（Rh-123）和抗体的荧光强度，评价APS对P-gp功能和表达的影响；建立Caco-2细胞单层模型，观察APS对Rh-123跨膜转运的影响。结果细胞毒性实验结果显示，APS在0.1~100μg/mL浓度范围内对Caco-2细胞无明显毒性，细胞存活率跃90%；不同浓度的APS作用后，增加细胞内Rh-123的蓄积，对P-gp功能表现为抑制作用；中、高浓度50、100μg/mL的APS对P-gp表达有抑制作用[（26.72±2.43）、（23.97±2.08）比（30.10±1.28），P〈0.05]，其表达量分别降低11.23%和16.41%；高浓度APS 100μg/mL明显抑制Rh-123的双向跨膜转运，对P-gp有抑制作用。结论黄芪多糖（APS）对P-gp的功能和表达有一定的抑制作用，且在高浓度尤为明显，能显著增加Rh-123在Caco-2细胞内的蓄积，并抑制其双向跨膜转运。
Objective To investigate the effect of astragalus polysaccharide（APS） on the function and expression of p-glycoprotein （P-gp） in Caco-2 cells. Methods The cytotoxicity of different drug concentrations was measured by MTT assay, in order to confirm the maximum non-toxic concentration of the drug. Flow cytometry was used to detect the fluorescence of rhodamine 123 concentration and antibody concentration to evaluate the effect of APS on the function and expression of P-gp. The Caco-2 cell monolayer was built to evaluate the effect of APS on the bi-directional transports of Rh-123. Results The cell viability of Caco-2 cells was higher than 90% when the concentration of APS was between 0.1 and 100μg/mL with MTT. Compared with control groups, APS increased the cellular Rh-123 concentration, showing an inhibitory effect on P-gp function. When the cells treated with APS at 50 and 100μg/mL concentrations, respectively, the expression levels of P-gp in Caco-2 cells were decreased by 11.23% and 16.41% to 26.72±2.43 and 23.97±2.08 compared to that in control group （30.10±1.28, P〈0.05）. Rh-123 bi-directional transport experiment showed that the p-gp level was obviously inhibited after 72 h incubation with 100 μg/mL concentration APS. Conclusion APS can inhibit the function and expression of P-gp in Caco-2 cells, and can also reduce the bi-directional transport of Rh-123 when it is at a high concentration.
出处
《浙江中西医结合杂志》
2015年第9期825-828,共4页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
基金
浙江省中西医结合学会科研项目(No.2013LYZD016)
