Endogenous bile acid disposition in rat and human sandwich-cultured hepatocytes

详细信息	查看全文 | 推荐本文 |

• 作者：Tracy L. Marion^c ; ¹ ; ^{tracylmarion@qualyst.com} ; Cassandra H. Perry^b ; ^{cassandraperry@qualyst.com} ; Robert L. St. Claire III^b ; ^{bobstclaire@qualyst.com} ; Kim L.R. Brouwer^a ; ^{kbrouwer@unc.edu}
• 关键词：BA ; bile acids ; BSEP ; bile salt export pump ; BEI ; biliary excretion index ; CTL ; control ; CYP450 ; cytochrome P450 ; DMSO ; dimethyl sulfoxide ; DMEM ; Dulbecco's modified Eagle medium ; EDTA ; ethylenediaminetetraacetic acid ; GCDCA ; glycochenodeoxycholic acid ; G
• 刊名：Toxicology and Applied Pharmacology
• 出版年：2012
• 期刊代码：58_0041008x
• 类别：pha
• 出版时间：15 May, 2012
• 卷：261
• 期：1
• 页码：1-9
• 文件大小：528 K

摘要

Sandwich-cultured hepatocytes (SCH) are used commonly to investigate hepatic transport protein-mediated uptake and biliary excretion of substrates. However, little is known about the disposition of endogenous bile acids (BAs) in SCH. In this study, four endogenous conjugated BAs common to rats and humans [taurocholic acid (TCA), glycocholic acid (GCA), taurochenodeoxycholic acid (TCDCA), and glycochenodeoxycholic acid (GCDCA)], as well as two BA species specific to rodents (伪- and 尾-tauromuricholic acid; 伪/尾 TMCA), were profiled in primary rat and human SCH. Using B-CLEAR^庐 technology, BAs were measured in cells + bile canaliculi, cells, and medium of SCH by LC-MS/MS. Results indicated that, just as in vivo, taurine-conjugated BA species were predominant in rat SCH, while glycine-conjugated BAs were predominant in human SCH. Total intracellular BAs remained relatively constant over days in culture in rat SCH. Total BAs in control (CTL) cells + bile, cells, and medium were approximately 3.4, 2.9, and 8.3-fold greater in human than in rat. The estimated intracellular concentrations of the measured total BAs were 64.3 卤 5.9 渭M in CTL rat and 183 卤 56 渭M in CTL human SCH, while medium concentrations of the total BAs measured were 1.16 卤 0.21 渭M in CTL rat SCH and 9.61 卤 6.36 渭M in CTL human SCH. Treatment of cells for 24 h with 10 渭M troglitazone (TRO), an inhibitor of the bile salt export pump (BSEP) and the Na⁺-taurocholate cotransporting polypeptide (NTCP), had no significant effect on endogenous BAs measured at the end of the 24-h culture period, potentially due to compensatory mechanisms that maintain BA homeostasis. These data demonstrate that BAs in SCH are similar to in vivo, and that SCH may be a useful in vitro model to study alterations in BA disposition if species differences are taken into account.