Proteomic analysis for the impact of hypercholesterolemia on expressions of hepatic drug transporters and metabolizing enzymes
Proteomic analysis for the impact of hypercholesterolemia on expressions of hepatic drug transporters and metabolizing enzymes
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高胆固醇血症对肝药物转运蛋白和代谢酶表达影响的蛋白质组学分析
DOI：
10.3109/00498254.2016.1144228
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发表时间：
2016-01-01
期刊：
影响因子：
1.8
通讯作者：
Yu, Chao
中科院分区：
文献类型：
--
作者：
Liu, Yan;Pu, Qiang-Hong;Yu, Chao
1.本研究旨在探讨高胆固醇血症时肝脏药物转运体和代谢酶的变化。给雄性Sprague-Dawley大鼠喂食高胆固醇饲料8周以诱导高胆固醇血症。采用iTRAQ标记结合LC-TRIPLE-TOF技术分析肝脏药物转运蛋白和代谢酶的蛋白水平。通过蛋白质组学分析，共鉴定出239个差异表达蛋白。其中，与对照组相比，高胆固醇血症组肝脏药物转运蛋白（MRP 2、ABCD 3、OAT 2、SLC 25 A12、SCL 38 A3、SLC 2A 2和SLC 25 A5）和代谢酶（CYP 2B 3、CYP 2C 7、CYP 2C 11、CYP 2C 13、CYP 4A 2和UGT 2B）的蛋白水平显著降低，而CYP 2C 6和CYP 2 E1的水平升高。通过真实的时间定量PCR（RT-qPCR）和western blot进一步证实了药物转运蛋白MRP 2和OAT 2的表达降低.免疫抑制通路分析表明，这些差异表达的蛋白质受到多种信号通路的调控，包括核受体和炎症细胞因子。通过RT-qPCR和蛋白质印迹进一步验证了核受体候选物之一，肝X受体α（LXR α）。此外，LXR α激动剂T0901317还能挽救高胆固醇血清处理后HepG 2细胞中MRP 2和OAT 2表达的降低.本研究结果表明，高胆固醇血症通过核受体途径影响肝脏多种药物转运体和代谢酶的表达。特别是，LXR α的功能降低有助于降低MRP 2和OAT 2的表达。
1. Our objective is to investigate the alterations of hepatic drug transporters and metabolizing enzymes in hypercholesterolemia. Male Sprague-Dawley rats were fed high-cholesterol chows for 8 weeks to induce hypercholesterolemia. Protein levels of hepatic drug transporters and metabolizing enzymes were analyzed by iTRAQ labeling coupled with LC TRIPLE-TOF.2. Total 239 differentially expressed proteins were identified using proteomic analysis. Among those, protein levels of hepatic drug transporters (MRP2, ABCD3, OAT2, SLC25A12, SCL38A3, SLC2A2 and SLC25A5) and metabolizing enzymes (CYP2B3, CYP2C7, CYP2C11, CYP2C13, CYP4A2 and UGT2B) were markedly reduced, but the levels of CYP2C6 and CYP2E1 were increased in hypercholesterolemia group compared to control. Decreased expressions of drug transporters MRP2 and OAT2 were further confirmed by real time quantitative PCR (RT-qPCR) and western blot.3. Ingenuity pathway analysis revealed that these differentially expressed proteins were regulated by various signaling pathways including nuclear receptors and inflammatory cytokines. One of the nuclear receptor candidates, liver X receptor alpha (LXRa), was further validated by RT-qPCR and western blot. Additionally, LXRa agonist T0901317 rescued the reduced expressions of MRP2 and OAT2 in HepG2 cells in hypercholesterolemic serum treatment.4. Our present results indicated that hypercholesterolemia affected the expressions of various drug transporters and metabolizing enzymes in liver via nuclear receptors pathway. Especially, decreased function of LXRa contributes to the reduced expressions of MRP2 and OAT2.