Robust protein nitration contributes to acetaminophen-induced mitochondrial dysfunction and acute liver injury.
Robust protein nitration contributes to acetaminophen-induced mitochondrial dysfunction and acute liver injury.
复制标题
DOI：
10.1016/j.freeradbiomed.2013.02.018
复制
发表时间：
2013-07
影响因子：
7.4
通讯作者：
Song, Byoung-Joon
中科院分区：
文献类型：
--
作者：
Abdelmegeed, Mohamed A.;Jang, Sehwan;Banerjee, Atrayee;Hardwick, James P.;Song, Byoung-Joon
关键词：
对乙酰氨基酚（APAP）是一种广泛使用的镇痛剂，可通过增加硝化应激导致蛋白质硝化而引起肝损伤。然而，硝化蛋白的身份及其在肝毒性中的作用知之甚少。因此，我们的目的是研究APAP诱导的肝毒性的机制，通过系统的鉴定和表征硝化蛋白质的存在或不存在的抗氧化剂N-乙酰半胱氨酸（NAC）。APAP单次给药（350 mg/kg ip）2 h后，小鼠肝脏硝化蛋白水平明显升高，24 h后肝脏出现严重坏死。在暴露于无毒3-羟基乙酰苯胺的小鼠或APAP和NAC联合治疗的动物中，蛋白质硝化和肝坏死最小。亲和纯化的硝化蛋白质的质谱分析确定了许多线粒体和胞质蛋白质，包括线粒体醛脱氢酶，锰超氧化物歧化酶，谷胱甘肽过氧化物酶，ATP合酶，和3-酮脂酰辅酶A硫解酶分别参与抗氧化防御，能量供应和脂肪酸代谢。免疫沉淀，然后用抗3-NT抗体免疫印迹证实，上述蛋白质被硝化在APAP暴露的小鼠，但不是在NAC共处理的小鼠。一致地，NAC共处理显著恢复了这些酶的抑制活性。因此，我们证明了一种新的机制，许多硝化蛋白与伴随抑制活动促进APAP诱导的线粒体功能障碍和肝毒性。
Acetaminophen (APAP), a widely-used analgesic agent, can cause liver injury through increased nitrative stress, leading to protein nitration. However, the identities of nitrated proteins and their roles in hepatotoxicity are poorly understood. Thus, we aimed at studying the mechanism of APAP-induced hepatotoxicity by systematic identification and characterization of nitrated proteins in the absence or presence of an anti-oxidant N-acetylcysteine (NAC). The levels of nitrated proteins markedly increased at 2 h in mice exposed to a single APAP dose (350 mg/kg ip), which caused severe liver necrosis at 24 h. Protein nitration and liver necrosis were minimal in mice exposed to nontoxic 3-hydroxyacetanilide or animals co-treated with APAP and NAC. Mass-spectral analysis of the affinity-purified nitrated proteins identified numerous mitochondrial and cytosolic proteins including mitochondrial aldehyde dehydrogenase, Mn-superoxide dismutase, glutathione peroxidase, ATP synthase, and 3-ketoacyl-CoA thiolase involved in anti-oxidant defense, energy supply, and fatty acid metabolism, respectively. Immunoprecipitation followed by immunoblot with anti-3-NT antibody confirmed that the aforementioned proteins were nitrated in APAP-exposed mice but not in NAC-co-treated mice. Consistently, NAC co-treatment significantly restored the suppressed activities of these enzymes. Thus, we demonstrate a new mechanism by which many nitrated proteins with concomitantly suppressed activities promotes APAP-induced mitochondrial dysfunction and hepatotoxicity.
登录
查看更多内容
影响因子：
4.1
作者：
Chen, Chi;Krausz, Kristopher W.;Shah, Yatrik M.;Idle, Jeffrey R.;Gonzalez, Frank J.
通讯作者：
Gonzalez, Frank J.
影响因子：
4.1
作者：
Hinson, JA;Pike, SL;Mayeux, PR
通讯作者：
Mayeux, PR
影响因子：
3.8
作者：
Jaeschke, H;Gores, GJ;Lemasters, JJ
通讯作者：
Lemasters, JJ
影响因子：
4.2
作者：
Abdelmegeed, Mohamed A.;Yoo, Seong-Ho;Song, Byoung-Joon
通讯作者：
Song, Byoung-Joon
影响因子：
13.5
作者：
Bourdi, M;Masubuchi, Y;Pohl, LR
通讯作者：
Pohl, LR