Clarification of xenobiotic recognition mechanism at the blood-placentabarrier by transporter array

转运阵列阐明血胎盘屏障的异生素识别机制

• 批准号: 18590154
• 负责人: SAI Yoshimichi
• 金额: $ 2.57万
• 依托单位: Kyoritsu University of Pharmacy
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590154/
• 关键词: Placenta barrier; Nucleoside; Nucleobase; Transporter; Drug Drug Interaction; Contribution; Anti-virus drug; recognition of xenobiotics; トロホブラスト; 妊娠時薬物療法; 有機アニオン; 定量PCR; Caco-2細胞; エストロン

The uptake mechanisms of nucleosides at the blood-placenta barrier were clarified by using the rat syncytiotrophoblast cell line TR-TBT 18d-1. The initial uptake of [3H]uridine and [3H]adenosine by TR-TBT 18d-1 were sodium-independent and were sensitive to nitrobenzylthioinosine. The Km value of adenosine uptake was approximately 17 μM. These results suggested involvement of equilibrative nucleoside transporters ENT1 and ENT2. The uptakes were significantly reduced by several nucleoside drugs, including zidovudine (AZT), cytarabine, vidarabine, mizoribine and caffeine. The effects were small within the therapeutic concentration ranges. Therefore, these drugs at the therapeutic concentrations might have little influence on maternal-to-fetal nucleoside transfer. The uptake mechanism of AZT was characterized. Initial uptake of AZT was sodium-independent and saturable (Km, approx. 16 μM). Thymidine and 2' -deoxyuridine strongly inhibited the uptake, but nitrobenzylthioinosine, probenecid a … More nd cimetidine had little effect on the uptake. Cyclosporin A, Ko143 and probenecid had little effect on the steady state AZT accumulation, suggesting that transporter-mediated efflux of AZT is not substantial. These results indicate that the saturable AZT uptake by TR-TBT 18d-1 was mediated by a so-far unidentified transporter.A human enterocyte model Caco-2 cell exhibits estrone-3-sulfate uptake, but the responsible transporters have not been clarified, yet. We have examined the contribution of OATP2B1 to the uptake in comparison with that of its homolog genes. The OATP2B1 mRNA expression was higher than that of OATP3A1 or OATP4A1 in Caco-2 and in human jejunum biopsies. The estrone-3-sulfate uptakes normalized to OATP2B1 mRNA expression were similar in Caco-2 cells and HEK293/OATP2B1 cells. The specific activity of OATP2B1 per mRNA expression was much higher than that of OATP3A1 and OATP4A1. These results suggest that OATP2B1 is predominantly responsible for the uptake in Caco-2 cells. This method can be applied to determine the contribution of each transporter in the placenta barrier. Less

利用大鼠合胞滋养细胞TR-TBT 18d-1，阐明了核苷在血-胎盘屏障中的摄取机制。TR-TBT 18d-1对[3H]尿苷和[3H]腺苷的初始摄取不依赖于钠，对硝基苄基硫代肌苷敏感。腺苷摄取Km值约为17 μM。这些结果表明参与了平衡核苷转运蛋白ENT1和ENT2。几种核苷类药物，包括齐多夫定（AZT）、阿糖胞苷、阿糖腺苷、米佐利滨和咖啡因，显著减少了摄取。在治疗浓度范围内，效果很小。因此，这些药物在治疗浓度下可能对母胎核苷转移影响不大。表征了AZT的吸收机理。AZT的初始摄取不依赖于钠，并且是饱和的(Km，大约。16μM)。胸腺嘧啶和2′-脱氧尿嘧啶对其吸收有较强的抑制作用，而硝基苄基硫代肌苷、丙戊酸、丙戊酸和西咪替丁对其吸收的影响较小。环孢素A、Ko143和probenecid对AZT稳态积累影响不大，提示转运体介导的AZT外排不明显。这些结果表明，TR-TBT 18d-1的饱和AZT摄取是由一种迄今尚未确定的转运体介导的。人肠细胞模型Caco-2细胞表现出雌酮-3-硫酸摄取，但负责的转运蛋白尚未明确。我们研究了OATP2B1对摄取的贡献，并与它的同源基因进行了比较。在Caco-2和人空肠活检组织中，OATP2B1 mRNA的表达高于OATP3A1或OATP4A1。在Caco-2细胞和HEK293/OATP2B1细胞中，雌激素-3-硫酸盐摄取与OATP2B1 mRNA表达归一化相似。每mRNA表达的OATP2B1比活性远高于OATP3A1和OATP4A1。这些结果表明OATP2B1主要负责Caco-2细胞的摄取。该方法可用于确定每个转运体在胎盘屏障中的贡献。少

项目成果

ラット胎盤細胞株を用いたヌクレオシド/ヌクレオベースの母体-胎児血輸送における薬物の相互作用解析.

使用大鼠胎盘细胞系分析核苷/核碱基母胎血液转运中的药物相互作用。

• 发表时间: 2007
• 作者: 地主 拓也;崔 吉道;巨勢 典子;佐藤 和子;西村 友宏;中島 恵美.
• 通讯作者: 中島 恵美.

薬物輸送駆動力解明に用いるラット胎盤刷子縁膜小胞の方向性及び精製度評価

评估大鼠胎盘刷状缘膜囊泡的方向和纯度，用于阐明药物转运的驱动力

• 发表时间: 2007
• 作者: 高木 彰紀;西村 友宏;崔 吉道;中島 恵美
• 通讯作者: 中島 恵美

Characterization of the placental transport mechanism of erythromycin using syncytiotrophoblast cell line TR-TBT 18d-1

使用合体滋养层细胞系 TR-TBT 18d-1 表征红霉素的胎盘转运机制

• 发表时间: 2007
• 作者: Nishimura T;Sai Y;Ochi K;Kose N;Nakashima E.
• 通讯作者: Nakashima E.

The expression levels of ERM during pregnancy and role of ezrin in localization of GLUT1 and P-glycoprotem in placenta

妊娠期ERM的表达水平及ezrin在胎盘GLUT1和P-糖蛋白定位中的作用

• 发表时间: 2007
• 作者: Higuchi K;Wada M;Deguchi M;Horieya S;Kose N;Nishimura T;Sai Y;Wakayama T;Tamura A;Tsukita S;Nakashima E.
• 通讯作者: Nakashima E.

Involvement of nucleoside transporters in the uptake of anti-HIV drugs,2',3'-dideoxyinosine and 3'-azido-3'-deoxythinudine by a conditionally immortalized syncytiotrophoblast cell line,TR-TBT

核苷转运蛋白参与条件永生化合体滋养层细胞系 TR-TBT 摄取抗 HIV 药物、2,3-二脱氧肌苷和 3-叠氮基-3-脱氧替尼啶

• 发表时间: 2007
• 作者: Nisimura T;Sato K;Sai Y;Kose N;Shimpo S;Nakashima E.
• 通讯作者: Nakashima E.