浙江中西医结合杂志
浙江中西醫結閤雜誌
절강중서의결합잡지
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
2015年
9期
825-828
,共4页
陈琳瑶%阮丹%王跃鑫%黄巧玲
陳琳瑤%阮丹%王躍鑫%黃巧玲
진림요%원단%왕약흠%황교령
黄芪多糖%P糖蛋白%Caco-2细胞
黃芪多糖%P糖蛋白%Caco-2細胞
황기다당%P당단백%Caco-2세포
astragalus polysaccharide%p-glycoprotein%Caco-2 cells
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。
目的：觀察黃芪多糖（APS）對Caco-2細胞P糖蛋白（P-gp）功能、錶達的影響。方法採用MTT法攷察藥物對細胞的毒性,確定藥物最大無毒濃度；流式細胞儀測定Caco-2細胞P-gp底物囉丹明-123（Rh-123）和抗體的熒光彊度，評價APS對P-gp功能和錶達的影響；建立Caco-2細胞單層模型，觀察APS對Rh-123跨膜轉運的影響。結果細胞毒性實驗結果顯示，APS在0.1~100μg/mL濃度範圍內對Caco-2細胞無明顯毒性，細胞存活率躍90%；不同濃度的APS作用後，增加細胞內Rh-123的蓄積，對P-gp功能錶現為抑製作用；中、高濃度50、100μg/mL的APS對P-gp錶達有抑製作用[（26.72±2.43）、（23.97±2.08）比（30.10±1.28），P<0.05]，其錶達量分彆降低11.23%和16.41%；高濃度APS 100μg/mL明顯抑製Rh-123的雙嚮跨膜轉運，對P-gp有抑製作用。結論黃芪多糖（APS）對P-gp的功能和錶達有一定的抑製作用，且在高濃度尤為明顯，能顯著增加Rh-123在Caco-2細胞內的蓄積，併抑製其雙嚮跨膜轉運。
목적：관찰황기다당（APS）대Caco-2세포P당단백（P-gp）공능、표체적영향。방법채용MTT법고찰약물대세포적독성,학정약물최대무독농도；류식세포의측정Caco-2세포P-gp저물라단명-123（Rh-123）화항체적형광강도，평개APS대P-gp공능화표체적영향；건립Caco-2세포단층모형，관찰APS대Rh-123과막전운적영향。결과세포독성실험결과현시，APS재0.1~100μg/mL농도범위내대Caco-2세포무명현독성，세포존활솔약90%；불동농도적APS작용후，증가세포내Rh-123적축적，대P-gp공능표현위억제작용；중、고농도50、100μg/mL적APS대P-gp표체유억제작용[（26.72±2.43）、（23.97±2.08）비（30.10±1.28），P<0.05]，기표체량분별강저11.23%화16.41%；고농도APS 100μg/mL명현억제Rh-123적쌍향과막전운，대P-gp유억제작용。결론황기다당（APS）대P-gp적공능화표체유일정적억제작용，차재고농도우위명현，능현저증가Rh-123재Caco-2세포내적축적，병억제기쌍향과막전운。
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.
