The effects of protease inhibitors and nonnucleoside reverse transcriptase inhibitors on P-glycoprotein expression in peripheral blood mononuclear cells in vitro
The effects of protease inhibitors and nonnucleoside reverse transcriptase inhibitors on P-glycoprotein expression in peripheral blood mononuclear cells in vitro
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DOI：
10.1097/00126334-200308150-00001
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发表时间：
2003-08-15
影响因子：
3.6
通讯作者：
Back, D
中科院分区：
文献类型：
--
作者：
Chandler, B;Almond, L;Back, D
几种抗逆转录病毒化合物已被证明是外排蛋白P-糖蛋白(P-gp)的底物，尽管很少有研究调查药物对该蛋白表达的影响。本研究采用体外实验系统，研究了蛋白水解酶抑制剂(PI)和非核苷类逆转录酶抑制剂(NNRTI)对外周血单个核细胞(PBMC)P-gp表达的影响。从健康志愿者分离的PBMC分别与10或100 MU的PI(沙奎那韦、利托那韦、洛比那韦、依地那韦、奈非那韦、氨丙那韦)或10 MU的NNRTI(Eefavirenz、奈韦拉平)孵育72小时。用流式细胞仪检测细胞表面P-gp的表达，并与载体孵育的对照组进行比较。用四甲基偶氮唑盐比色法(MTT法)评价各化合物的毒性，并比较MDR1基因外显子26第3435位不同基因型个体对各化合物的影响。Nelfinavir(10.2个P-gp阳性细胞)和efavirenz(10.0个P-gp阳性细胞，6.7%P-gp阳性细胞)孵育后，P-gp表达中位数显著增加。不同基因型(CC、CT、TT)间的诱导效果无显著差异。与100um PI孵育后，除氨丙那韦外，P-gp显著上调。然而，奈非那韦、利托那韦和洛比那韦引起了明显的毒性，表明在较高浓度下，P-gp的增加可能至少部分与应激反应有关。这些结果表明，一些PI和NNRTI在体外可以诱导PBMCs表达P-gp。
Several antiretroviral compounds have been shown to be substrates for the efflux protein P-glycoprotein (P-gp) although few studies have investigated the effects of drug on expression of this protein. Here, an in vitro system has been adopted to investigate the effects of protease inhibitors (PIs) and nonnucleoside reverse transcriptase inhibitors (NNRTIs) on P-gp expression in peripheral blood mononuclear cells (PBMCs). PBMCs isolated from healthy volunteers were incubated with 10 or 100 muM PI (saquinavir, ritonavir, lopinavir, indinavir, nelfinavir, amprenavir) or 10 muM NNRTI (efavirenz, nevirapine) for 72 hours. Surface P-gp expression was measured by flow cytometry and compared with vehicle-incubated controls. Toxicity was assessed by MTT assay and the effects of each compound were compared between individuals with differing genotypes at position 3435 of exon 26 of MDR1, which was assigned by restriction fragment length polymorphism. Significant increases in median P-gp expression were observed following incubation with 10 muM nelfinavir (10.2 versus 6.7% P-gp-positive cells) and efavirenz (10.0 versus 6.7% P-gp-positive cells). No significant differences in induction were observed between genotypes (CC, CT, TT). Following incubation with 100 muM PI, significant upregulation of P-gp occurred except with amprenavir. However, nelfinavir, ritonavir, and lopinavir caused marked toxicity, indicating that at higher concentrations, the increase in P-gp may be at least partially related to a stress response. These results indicate the potential of some PIs and NNRTIs to induce P-gp expression in PBMCs in vitro.