摘要
目的 研究普流罗尼克（Pluronic F127，F68，P85，P123对小肠P-糖蛋白（P-gp的调控作用。方法 采用MTT法检测不同质量浓度的Pluronic对Caco-2细胞生长抑制作用；以P-gp底物罗丹明123(R-123为荧光探针，评价各种Pluronic辅料和P-gp抑制剂维拉帕米对R-123细胞蓄积的影响，细胞内的R-123浓度采用高效液相-荧光检测。同时考察了Pluronic P123和F127对P-gp ATP酶活性的影响。结果 除了L61，经Pluronic处理后的细胞生存率都在80％以上，表明在蓄积实验中的细胞活性没有受到Pluronic的影响。在抑制剂维拉帕米和不同浓度的Pluronic(0.001~50 mg·mL-1作用下，可在不同程度上提高R-123在Caco-2细胞的蓄积作用，同时具有浓度依赖性，在接近或超过临界胶束浓度(CMC后，细胞蓄积达到最大值，然后随着Pluronic浓度的继续增大，蓄积作用又逐渐减弱。不同浓度的Pluronic P123和F127对P-gp ATP酶活性具有抑制作用。结论 蓄积实验结果表明，Pluronic F127，F68，P85和P123可以通过抑制P-gp的作用改善药物的吸收，抑制P-gp ATP酶活性可能是原因之一。因此，联合使用Pluronic，有望提高P-gp底物药物的口服生物利用度。
Abstract
OBJECTIVE To determine whether Pluronic， P123， F127， P85， F68 and L61 possess ability to modulate P-glycoprotein(P-gp mediated efflux of drugs. METHODS Caco-2 cell viability was measured using MTT assay in the presence of Pluronic at different concentrations. To evaluate the effects of Pluronic and verapamil（inhibitor of P-gpon cellular drug accumulation in caco-2 cells， rhodamine 123(R-123， a P-gp substrate， was selected as fluorescent probe. A HPLC-fluorescence method was used to determined the concentration of R-123 in Caco-2 cells. Caco-2 cell membranes were exposed to various Pluronic solutions（P123 or F127， and then the ATPase activity of P-gp was assayed by determining liberated inorganic phosphate. RESULTS The viability of Pluronic-treated cells was above 80% except L61， which suggested that Caco-2 cells could maintain their viability under the experiment conditions. In the presence of verapamil and Pluronic at concentrations ranging from 0.00 1-50 mg·mL-1， R-123 accumulation were increased significantly to differernt extent. R-123 accumulation reached maximum levels when Pluronic concentrations were close to or above the CMC， Higher Pluronic concentrations resulted in a drop in accumulation compared to that of R-123 control. Furthermore， P123 and F127 treatments decreased P-gp ATPase activity in Caco-2 cell membranes expressing P-gp. CONCLUSION Pluronic(F68， F127， P123 and P85 can modulate drug efflux by inhibiting P-gp activity in Caco-2 cell membranes. It may be possible to improve the oral bioavailability of P-gp substrates by co-administration of Pluronic.
关键词
Pluronic /
P-糖蛋白 /
罗丹明123 /
Caco-2细胞
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Key words
Pluronic /
P-gp /
rhodamine 123 /
Caco-2 cell
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关延彬 左岚 饶子超 斯陆勤 李高.
普流罗尼克对P-gp底物罗丹明123在Caco-2细胞蓄积的影响[J]. 中国药学杂志, 2011, 46(12): 937-942
GUN Yn-in;ZUO Ln;RO Zi-cho;SI Lu-qin;LI Go.
Effect of Pluronic on Cellular Accumulation of Rhodamine 123， a P-gp Substrate in Caco-2 Cells[J]. Chinese Pharmaceutical Journal, 2011, 46(12): 937-942
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