Filipović Dragana, Costina Victor, Perić Ivana, Stanisavljević Andrijana, Findeisen Peter
Vinča Institute of Nuclear Sciences, Laboratory for Molecular Biology and Endocrinology, University of Belgrade, Serbia. Electronic address: http://www.vinca.rs.
Institute for Clinical Chemistry, Medical Faculty Mannheim of the University of Heidelberg, University Hospital Mannheim, Germany.
Brain Res. 2017 Mar 15;1659:41-54. doi: 10.1016/j.brainres.2017.01.025. Epub 2017 Jan 21.
Fluoxetine (Flx) is the principal treatment for depression; however, the precise mechanisms of its actions remain elusive. Our aim was to identify protein expression changes within rat hippocampus regulated by chronic Flx treatment versus vehicle-controls using proteomics. Fluoxetine-hydrohloride (15mg/kg) was administered daily to adult male Wistar rats for 3weeks, and cytosolic and nonsynaptic mitochondrial hippocampal proteomes were analyzed. All differentially expressed proteins were functionally annotated according to biological process and molecular function using Uniprot and Blast2GO. Our comparative study revealed that in cytosolic and nonsynaptic mitochondrial fractions, 60 and 3 proteins respectively, were down-regulated, and 23 and 60 proteins, respectively, were up-regulated. Proteins differentially regulated in cytosolic and nonsynaptic mitochondrial fractions were primarily related to cellular and metabolic processes. Of the identified proteins, the expressions of calretinin and parvalbumine were confirmed. The predominant molecular functions of differentially expressed proteins in both cell hippocampal fractions were binding and catalytic activity. Most differentially expressed proteins in nonsynaptic mitochondria were catalytic enzymes involved in the pyruvate metabolism, citric acid cycle, oxidative phosphorylation, ATP synthesis, ATP transduction and glutamate metabolism. Results indicate that chronic Flx treatment may influence proteins involved in calcium signaling, cytoskeletal structure, chaperone system and stimulates energy metabolism via the upregulation of GAPDH expression in cytoplasm, as well as directing energy metabolism toward the citric acid cycle and oxidative phosphorylation in nonsynaptic mitochondria. This approach provides new insight into the chronic effects of Flx treatment on protein expression in a key brain region associated with stress response and memory.
氟西汀（Flx）是治疗抑郁症的主要药物；然而，其确切作用机制仍不清楚。我们的目的是通过蛋白质组学确定慢性氟西汀治疗组与溶剂对照组相比，大鼠海马体中受调控的蛋白质表达变化。将盐酸氟西汀（15mg/kg）每日给予成年雄性Wistar大鼠，持续3周，并分析海马体胞质和非突触线粒体蛋白质组。使用Uniprot和Blast2GO根据生物学过程和分子功能对所有差异表达的蛋白质进行功能注释。我们的比较研究表明，在胞质和非突触线粒体组分中，分别有60种和3种蛋白质下调，23种和60种蛋白质上调。在胞质和非突触线粒体组分中差异调节的蛋白质主要与细胞和代谢过程有关。在已鉴定的蛋白质中，钙视网膜蛋白和小白蛋白的表达得到了证实。海马体两个细胞组分中差异表达蛋白质的主要分子功能是结合和催化活性。非突触线粒体中大多数差异表达的蛋白质是参与丙酮酸代谢、柠檬酸循环、氧化磷酸化、ATP合成、ATP转导和谷氨酸代谢的催化酶。结果表明，慢性氟西汀治疗可能会影响参与钙信号传导、细胞骨架结构、伴侣系统的蛋白质，并通过上调细胞质中GAPDH的表达来刺激能量代谢，以及将能量代谢导向非突触线粒体中的柠檬酸循环和氧化磷酸化。这种方法为氟西汀治疗对与应激反应和记忆相关的关键脑区蛋白质表达的慢性影响提供了新的见解。
