Effect of hr-IL2 treatment on intestinal P-glycoprotein expression and activity in Caco-2 cells
Effect of hr-IL2 treatment on intestinal P-glycoprotein expression and activity in Caco-2 cells
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DOI：
10.1211/002235702320266262
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发表时间：
2002-08-01
影响因子：
3.3
通讯作者：
Leroy, C
中科院分区：
文献类型：
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作者：
Belliard, AM;Tardivel, S;Leroy, C
采用体外培养的Caco-2细胞，观察重组人白介素2(IL-2)对肠道P-糖蛋白(P-gp)转运体活性的影响。更具体地说，通过测量P-gp的荧光底物罗丹明-123的跨上皮转运来研究P-gp的外排功能。在我们的实验中，两种特异性P-gp抑制剂环孢素A和GG918完全抑制了它的转运。相反，当以[H-3]长春新碱为底物时，这两种特异性P-gp抑制剂仅抑制50%的跨上皮转运。100IU mL(-1)(6.1ngmL(-1))的IL2作用于Caco-2细胞24 h后，罗丹明-123底物上的P-gp转运蛋白功能明显下降(21%)。在暴露于IL2的48和72小时后，这一效应也得到证实。然而，对于较高浓度的IL2(1000和5000IU mL(-1))，P-gp功能只有在较长的治疗时间(48h或更长)后才会发生。IL2对P-gp活性的抑制作用不依赖于紧密连接功能，本研究中所有实验条件(IL2暴露时间和浓度)的跨上皮电阻(TEER)的恒定测量证实了这一点。此外，1000IU m L-1IL2作用72 h后，Caco-2细胞中mdr1基因的表达水平受到强烈抑制，而MRP1基因的表达水平无明显变化。结论：Caco-2细胞与IL-2急性孵育后，P-gp转运蛋白表达和活性下降。
Caco-2 cells were used to investigate the effect of human recombinant interleukin-2 (IL-2) on intestinal P-glycoprotein (P-gp) transporter activity in-vitro. More specifically the efflux function of P-gp was studied by measuring the transepithelial transport of rhodamine-123, a fluorescent substrate of P-gp. Its transport was completely inhibited by two specific P-gp inhibitors, ciclosporin A and GG918, in our experiments. Conversely, these two specific P-gp inhibitors inhibited only 50% of transepithelial transport when [H-3]vincristine was used as substrate. After Caco-2 cells were treated with 100 IU mL(-1) (6.1 ng mL(-1)) IL2 for 24 h, a significant diminution (21%) of P-gp transporter function was observed with rhodamine-123 substrate. This effect was also confirmed after 48 and 72 h of exposure to IL2. However, for higher concentrations of IL2 (1000 and 5000 IU mL(-1)), diminution of P-gp function only occurred after a longer treatment period (48 h and more). The inhibitory effect of IL2 on P-gp activity was found to be independent of tight junction function as demonstrated by constant transepithelial electrical resistance (TEER) measurements for all experimental conditions encountered in this study (time and concentration of IL2 exposure). Furthermore, the MDR1 mRNA level was found to be strongly repressed in Caco-2 cells exposed with 1000 IU mL-1 IL2 for 72 h while the amount of MRP1 mRNA remained unchanged. In conclusion, acute incubation of Caco-2 cells with IL2 induced a decrease of P-gp transporter expression and activity.