Functional characteristics of 3'-azido-3'-deoxythymidine transport at the blood-testis barrier.
Functional characteristics of 3'-azido-3'-deoxythymidine transport at the blood-testis barrier.
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DOI：
10.1016/j.ijpharm.2022.122044
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发表时间：
2022-07
影响因子：
5.8
通讯作者：
Takeru Ito;Yoshiyuki Kubo;S. Akanuma;K. Hosoya
中科院分区：
文献类型：
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作者：
Takeru Ito;Yoshiyuki Kubo;S. Akanuma;K. Hosoya
3′-叠氮-3 ′-脱氧胸苷（AZT）是治疗人类免疫缺陷病毒（HIV）感染的常用抗逆转录病毒药物，本研究探讨了AZT在血睾屏障（BTB）上的转运特性。在评价体内转运活性的积分图分析中，[3 H]AZT的表观内流清除率显著大于非渗透性细胞旁转运标记物[14 C] D-甘露醇。在小鼠睾丸支持细胞来源的TM 4细胞体外摄取研究中，[3 H]AZT的摄取呈时间和浓度依赖性，其Km_（max）分别为20.3 μM和102 pmol/（min·mg protein）。在抑制分析中，[3 H]AZT的摄取不受胞外无机物和参与AZT转运的转运蛋白purons的某些底物的影响。在进一步阐明AZT转运特征的抑制分析中，在存在几种核苷（归类为含嘧啶的2′-脱氧核苷）的情况下，[3 H]AZT摄取显著降低，而在存在其他核苷、核碱基和抗逆转录病毒药物的情况下，对[3 H]AZT摄取几乎没有影响。这些结果表明，AZT从循环血液进入睾丸的内流转运，并参与载体介导的过程中的BTB，它选择性地识别2′-脱氧核苷与嘧啶碱基。
3′-Azido-3′-deoxythymidine (AZT), an antiretroviral drug, is often adopted in the therapy for human immunodeficiency virus (HIV) infection, and the characteristics of AZT transport at the blood-testis barrier (BTB) were investigated in this study. In the integration plot analysis that evaluates the transport activityin vivo, the apparent influx clearance of [3H]AZT was significantly greater than that of [14C]D-mannitol, a non-permeable paracellular transport marker. In the uptake studyin vitrowith TM4 cells derived from mouse Sertoli cells, [3H]AZT uptake exhibited a time- and concentration-dependent manner, of whichKmandVmaxvalues being 20.3 µM and 102 pmol/(min·mg protein), respectively. In the inhibition analysis, [3H]AZT uptake was not affected by extracellular inorganics and some substrates of transporters putatively involved in AZT transport. In the further inhibition analyses to elucidate the characteristics of AZT transport, [3H]AZT uptake was strongly reduced in the presence of several nucleosides, that are categorized as 2′-deoxynucleosides with pyrimidine, whereas little effect on [3H]AZT uptake was exhibited in the presence of other nucleosides, nucleobases, and antiretrovirals. These results suggest the influx transport of AZT from the circulating blood to the testis, and the involvement of carrier-mediated process at the BTB, which selectively recognizes 2′-deoxynucleosides with a pyrimidine base.