Testosterone and androstenedione are endogenous substrates of P-glycoprotein
Testosterone and androstenedione are endogenous substrates of P-glycoprotein
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睾酮和雄烯二酮是 P-糖蛋白的内源性底物
DOI：
10.1016/j.bbrc.2019.09.067
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发表时间：
2019
影响因子：
3.1
通讯作者：
Ogihara Takuo
中科院分区：
文献类型：
--
作者：
Yano Kentaro;Seto Saeka;Kamioka Hiroki;Mizoi Kenta;Ogihara Takuo
脑内睾酮（Tes）水平升高以及导致转运功能受损的P-糖蛋白（P-gp）单核苷酸多态性与自杀风险增加有关。在这里，我们研究了Tes及其前体和代谢产物是否是P-gp的底物，使用几种体外方法。在ATP酶测定中，随着Tes、二氢表雄酮（Dhea）、雄烯二酮（Ado）和二氢睾酮（Dht）浓度的增加，观察到ATP消耗增加，但不观察到雄烯二醇（Adol），这表明这四种雄激素由P-gp转运。此外，Tes和Ado，而不是Dhea或Dht，增加了P-gp过表达细胞系中P-gp典型底物罗丹明123（Rho 123）的细胞内蓄积，表明它们抑制Rho 123外排，因此是P-gp的底物或抑制剂。在Transwell插入物中使用P-gp过表达细胞进行的膜渗透性研究表明，Ado和Tes在基底至顶端方向（排泄）的渗透系数显著高于顶端至基底方向的渗透系数。此外，Ado和Tes的转运被典型的P-gp抑制剂维拉帕米显著抑制。这些结果表明，Tes和Ado是P-gp的内源性底物。这些发现为理解先前报道的P-gp功能障碍和Tes脑水平升高与自杀行为的相关性提供了生理学基础，并可能为治疗有自杀风险的患者开辟新的可能性。
Raised brain levels of testosterone (Tes), as well as single nucleotide polymorphisms of P-glycoprotein (P-gp) that cause impaired transport function, are associated with increased risk of suicide. Here, we examined whether Tes and its precursors and metabolites are substrates of P-gp, using several in vitro methods. In ATPase assay, increased ATP consumption was observed as the concentrations of Tes, dihydroepiandrosterone (Dhea), androstenedione (Ado), and dihydrotestosterone (Dht), but not androstenediol (Adol), were increased, suggesting that these four androgens are transported by P-gp. Furthermore, Tes and Ado, though not Dhea or Dht, increased the intracellular accumulation of Rhodamine 123 (Rho123), a typical substrate of P-gp, in a P-gp-overexpressing cell line, suggesting that they inhibit Rho123 efflux and thus are substrates or inhibitors of P-gp. A membrane permeability study using P-gp-overexpressing cells in Transwell inserts indicated that the permeability coefficients of both Ado and Tes in the basal-to-apical direction (excretion) are significantly higher than those in the apical-to-basal direction. Moreover, transport of both Ado and Tes was significantly suppressed by verapamil, a typical P-gp inhibitor. These results indicate that Tes and Ado are endogenous substrates of P-gp. These findings provide a physiological basis for understanding previously reported associations of P-gp dysfunction and raised brain levels of Tes with suicidal behavior, and may open up new possibilities for treating patients at risk of suicide.