Troglitazone-induced hepatic mitochondrial proteome expression dynamics in heterozygous Sod2+/- mice:: Two-stage oxidative injury
Troglitazone-induced hepatic mitochondrial proteome expression dynamics in heterozygous Sod2+/- mice:: Two-stage oxidative injury
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DOI：
10.1016/j.taap.2008.03.025
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发表时间：
2008-08-15
影响因子：
3.8
通讯作者：
Boelsterli, Urs A.
中科院分区：
文献类型：
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作者：
Lee, Yie Hou;Chung, Maxey C. M.;Boelsterli, Urs A.
尚未确定对曲格列酮诱导的特异质肝损伤易感性的决定因素;然而，曲格列酮已被证明靶向线粒体并在体外诱导线粒体介导的肝细胞损伤。本研究的目的是使用系统的方法来分析蛋白质组水平上的线粒体变化的动态，并通过使用先前表征的对曲格列酮肝毒性高度致敏的小鼠模型来更清楚地定义曲格列酮肝毒性的机制和时程。对线粒体超氧化物歧化酶-2（Sod 2（+/-））杂合子小鼠每天腹腔注射曲格列酮（30 mg/kg/天）或溶媒，持续2或4周。肝线粒体分离，纯化，并进行二维差异凝胶电泳（2D-DIGE）。我们发现，在类似的1500解决肝线粒体蛋白，70表现出显着改变后，曲格列酮治疗丰度。MALDI-TOF/TOF MS/MS分析显示，早期变化（2周）包括热休克蛋白家族成员（死亡蛋白，HSP 7 C），Lon蛋白酶和过氧化氢酶的水平增加，表明诱导线粒体应激反应。相反，4周后，一些关键蛋白质，包括ATP合酶β亚基，乌头酸酶-2，和过氧化氢酶表现出减少的丰度，和总蛋白质羰基显着增加，表明未补偿的氧化损伤。在这两个时间点，腺苷三磷酸酶-2（ACO 2）均降低，使该蛋白成为线粒体氧化应激的潜在敏感和早期生物标志物。这些结果表明，在这种潜在的临床沉默线粒体应激的小鼠模型中，叠加曲格列酮诱导两阶段反应：初始适应性反应，随后是涉及线粒体蛋白氧化损伤的毒性反应。(c)2008年爱思唯尔公司All rights reserved.
The determinants of susceptibility to troglitazone-induced idiosyncratic liver injury have not yet been determined; however, troglitazone has been shown to target mitochondria and induce mitochondria-mediated hepatocellular injury in vitro. The aim of this study was to use a systems approach to analyze the dynamics of mitochondrial changes at the proteome level and more clearly define the mechanisms and time course of troglitazone hepatotoxicity by using a previously characterized mouse model that is highly sensitized to troglitazone hepatotoxicity. Mice heterozygous in mitochondrial superoxide dismutase-2 (Sod2(+/-)) were injected intraperitoneally with troglitazone (30 mg/kg/day) or vehicle daily for 2 or 4 weeks. Hepatic mitochondria were isolated, purified, and subjected to two-dimensional difference gel electrophoresis (2D-DIGE). We found that among the similar to 1500 resolved hepatic mitochondrial proteins, 70 exhibited significantly altered abundance after troglitazone treatment. MALDI-TOF/TOF MS/MS analysis revealed that early changes (2 weeks) included increased levels of heat shock protein family members (mortalin, HSP7C), Lon protease, and catalase, indicating induction of a mitochondrial stress response. In contrast, after 4 weeks, a number of critical proteins including ATP synthase beta-subunit, aconitase-2, and catalase exhibited decreased abundance, and total protein carbonyls were significantly increased, suggesting uncompensated oxidative damage. Aconitase-2 (ACO2) was decreased at both time points, making this protein a potential sensitive and early biomarker for mitochondrial oxidant stress. These results show that, in this murine model of underlying clinically silent mitochondrial stress, superimposed troglitazone induces a two-stage response: an initial adaptive response, followed by a toxic response involving oxidant injury to mitochondrial proteins. (c) 2008 Elsevier Inc. All rights reserved.