Inhibition of Na+-Taurocholate Co-transporting Polypeptide-mediated Bile Acid Transport by Cholestatic Sulfated Progesterone Metabolites
Inhibition of Na+-Taurocholate Co-transporting Polypeptide-mediated Bile Acid Transport by Cholestatic Sulfated Progesterone Metabolites
复制标题
DOI：
10.1074/jbc.m109.072140
复制
发表时间：
2010-05-28
影响因子：
4.8
通讯作者：
Williamson, Catherine
中科院分区：
文献类型：
--
作者：
Abu-Hayyeh, Shadi;Martinez-Becerra, Pablo;Williamson, Catherine
硫酸孕酮代谢物（P4-S）水平在正常妊娠中升高，在妊娠肝内胆汁淤积症（ICP）中进一步升高，这是一种妊娠胆汁酸-肝脏疾病。ICP可因早产和宫内死亡而复杂化。采用培养的原代人肝细胞（PHH）和表达Na+-牛磺胆酸共转运多肽（NTCP）的非洲爪蟾卵母细胞两种肝脏胆汁酸摄取实验模型，研究P4-S对胆汁酸摄取的影响。两种P4-S化合物，异孕酮-硫酸盐（PM4-S）和表异孕酮-硫酸盐（PM5-S），在PHH中以剂量依赖的方式减少[H-3]牛磺胆酸（TC）的摄取，Na+依赖性和非依赖性胆汁酸摄取系统均被显著抑制。PM5-S介导的TC摄取抑制可以通过增加TC浓度来逆转，而PM5-S剂量固定表明竞争性抑制。利用表达ncpp的爪蟾卵母细胞进行的实验证实，PM4-S/PM5-S能够竞争性地抑制ncpp介导的[H-3]TC摄取。采用液相色谱-电喷雾串联质谱法测定未怀孕和妊娠晚期孕妇血清PM4-S + PM5-S水平，孕妇血清PM4-S + PM5-S水平升高，达到能够抑制TC摄取的水平。综上所述，妊娠期P4-S水平可抑制PHH中Na+依赖性和非依赖性牛磺胆酸内流，并竞争性抑制ncp介导的爪蟾卵母细胞对牛磺胆酸的摄取。
Sulfated progesterone metabolite (P4-S) levels are raised in normal pregnancy and elevated further in intrahepatic cholestasis of pregnancy (ICP), a bile acid-liver disorder of pregnancy. ICP can be complicated by preterm labor and intrauterine death. The impact of P4-S on bile acid uptake was studied using two experimental models of hepatic uptake of bile acids, namely cultured primary human hepatocytes (PHH) and Na+-taurocholate co-transporting polypeptide (NTCP)-expressing Xenopus laevis oocytes. Two P4-S compounds, allopregnanolone-sulfate (PM4-S) and epiallopregnanolone-sulfate (PM5-S), reduced [H-3] taurocholate (TC) uptake in a dose-dependent manner in PHH, with both Na+-dependent and-independent bile acid uptake systems significantly inhibited. PM5-S-mediated inhibition of TC uptake could be reversed by increasing the TC concentration against a fixed PM5-S dose indicating competitive inhibition. Experiments using NTCP-expressing Xenopus oocytes confirmed that PM4-S/PM5-S are capable of competitively inhibiting NTCP-mediated uptake of [H-3]TC. Total serum PM4-S + PM5-S levels were measured in non-pregnant and third trimester pregnant women using liquid chromatography-electrospray tandem mass spectrometry and were increased in pregnant women, at levels capable of inhibiting TC uptake. In conclusion, pregnancy levels of P4-S can inhibit Na+-dependent and-independent influx of taurocholate in PHH and cause competitive inhibition of NTCP-mediated uptake of taurocholate in Xenopus oocytes.