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长期暴露于氟化物的大鼠肾脏的蛋白质组学分析

Proteomic analysis of kidney in rats chronically exposed to fluoride.

作者信息

Kobayashi Claudia A N, Leite Aline L, Silva Thelma L, Santos Lucilene D, Nogueira Fábio C S, Oliveira Rodrigo C, Palma Mario S, Domont Gilberto B, Buzalaf Marília A R

机构信息

Bauru Dental School, University of São Paulo, Bauru, SP, Brazil.

出版信息

Chem Biol Interact. 2009 Jul 15;180(2):305-11. doi: 10.1016/j.cbi.2009.03.009. Epub 2009 Mar 24.

Abstract

Two-dimensional gel electrophoresis (2-DE) was used to better understand alterations in renal metabolism induced by fluoride (F). Three groups of weanling male Wistar rats were treated with drinking water containing 0 (control), 5, or 50 ppm F for 60 days (n=6/group). Kidneys were collected for proteomic and histological (HE) analysis. After protein isolation, renal proteome profiles were examined using 2-DE and Colloidal Coomassie Blue staining. Protein spots with a 2-fold significant difference as detected by quantitative intensity analysis (Image Master Platinum software) and t-test (p<0.05) were excised and analyzed by MALDI-TOF MS (matrix assisted laser desorption ionization-time-of-flight mass spectrometry). The histological analysis revealed no damage in kidneys induced by F, except for a vascular congestion in the 50 ppm F group. Between control vs 50 ppm F, and control vs 5 ppm F groups, 12 and 6 differentially expressed proteins were detected, respectively. Six proteins, mainly related with metabolism, detoxification and housekeeping, were successfully identified. At the high F group, pyruvate carboxylase, a protein involved in the formation of oxaloacetate was found to be downregulated, while enoyl coenzyme A hydratase, involved in fatty acids oxidation, was found to be upregulated. Thus, proteomic analysis can provide new insights into the alterations in renal metabolism after F exposure, even in low doses.

摘要

采用二维凝胶电泳(2-DE)以更好地了解氟(F)诱导的肾脏代谢变化。将三组断乳雄性Wistar大鼠分别用含0(对照)、5或50 ppm氟的饮用水处理60天(每组n = 6)。收集肾脏进行蛋白质组学和组织学(苏木精-伊红染色,HE)分析。蛋白质分离后,使用2-DE和考马斯亮蓝染色检查肾脏蛋白质组图谱。通过定量强度分析(Image Master Platinum软件)和t检验(p<0.05)检测到具有2倍显著差异的蛋白质斑点被切除,并通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)进行分析。组织学分析显示,除50 ppm氟组出现血管充血外,氟未对肾脏造成损伤。在对照与50 ppm氟组之间以及对照与5 ppm氟组之间,分别检测到12种和6种差异表达蛋白。成功鉴定出6种主要与代谢、解毒和管家功能相关的蛋白质。在高氟组中,参与草酰乙酸形成的丙酮酸羧化酶被发现下调,而参与脂肪酸氧化的烯酰辅酶A水合酶被发现上调。因此,蛋白质组学分析可为氟暴露后肾脏代谢变化提供新的见解,即使是低剂量暴露。

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