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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。
Objective To study the regulation of P-glycoprotein (P-gp) by Pluronic F127, F68, P85 and P123. Methods MTT assay was used to determine the inhibitory effect of Pluronic on the proliferation of Caco-2 cells. P-gp inhibitor Rhodamine 123 (R-123) was used as fluorescent probe to evaluate the effects of various Pluronic excipients and P-gp inhibitors Lapatin on the accumulation of R-123 cells, intracellular concentration of R-123 using high-performance liquid-fluorescence detection. The effects of Pluronic P123 and F127 on P-gp ATPase activity were also investigated. Results With the exception of L61, cell viability after Pluronic treatment was above 80%, indicating that the cell viability in the accumulation experiment was not affected by Pluronic. The inhibitory effect of verapamil and different concentrations of Pluronic (0.001 ~ 50 mg.mL-1) increased the accumulation of R-123 in Caco-2 cells to a certain extent and in a concentration-dependent manner. At or near the critical micelle concentration (CMC), cell accumulation reached its maximum, and then with the Pluronic concentration continues to increase, the accumulation gradually weakened. Different concentrations of Pluronic P123 and F127 have an inhibitory effect on P-gp ATPase activity. Conclusion Accumulation experiments showed that Pluronic F127, F68, P85 and P123 could improve the drug absorption and inhibit the activity of P-gp ATPase by inhibiting the action of P-gp. Therefore, the combination of Pluronic, is expected to increase the oral bioavailability of P-gp substrate drugs.