Reactive nitrogen species in acetaminophen-induced mitochondrial damage and toxicity in mouse hepatocytes.
Reactive nitrogen species in acetaminophen-induced mitochondrial damage and toxicity in mouse hepatocytes.
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DOI：
10.1021/tx1001755
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发表时间：
2010-07-19
影响因子：
4.1
通讯作者：
Hinson, Jack A.
中科院分区：
文献类型：
--
作者：
Burke, Angela S.;MacMillan-Crow, Lee Ann;Hinson, Jack A.
对乙酰氨基酚（APAP）对小鼠原代肝细胞的毒性发生在两个阶段。初始阶段（0-2小时）发生在n -乙酰-对苯醌亚胺的代谢过程中，它消耗谷胱甘肽，并与蛋白质共价结合，但毒性很小。随后洗涤肝细胞以去除APAP并在培养基中单独孵育（2-5小时）导致毒性。我们之前报道过，孵育期发生在线粒体通透性转变（MPT）和氧化应激（二氯二氢荧光素荧光）（DCFH2）增加的过程中。由于DCFH2可能通过多种氧化机制被氧化，我们通过ELISA和免疫印迹研究了活性氮物种（RNS）在蛋白质中导致3-硝基酪氨酸的作用。APAP与肝细胞孵育2小时不产生毒性或蛋白质硝化；然而，洗涤肝细胞并单独在培养基中孵育（2-5h）会导致蛋白质硝化，这与毒性有关。在孵育培养基中加入MPT抑制剂环孢素A可消除毒性和蛋白质硝化。再培养液中加入一般一氧化氮合酶（NOS）抑制剂L-NMMA和神经元一氧化氮合酶（NOS1）抑制剂7-硝基茚唑可降低毒性和蛋白质硝化作用；然而，诱导型NOS （NOS2）抑制剂L-NIL （N6-（1-氨基乙基）-l-赖氨酸）和SAIT （S-（2-氨基乙基）异硫脲）均未降低蛋白质硝化或毒性。在孵育期添加RNS清除剂n -乙酰半胱氨酸和高浓度APAP可降低毒性和蛋白质硝化。7-硝基吲哚唑和环孢素A抑制apap诱导的线粒体膜电位损失。这些数据表明RNS在APAP诱导的毒性中起作用。
Acetaminophen (APAP) toxicity in primary mouse hepatocytes occurs in two phases. The initial phase (0–2 h) occurs with metabolism to N-acetyl-p-benzoquinoneimine which depletes glutathione, and covalently binds to proteins, but little toxicity is observed. Subsequent washing of hepatocytes to remove APAP and reincubating in media alone (2–5 h) results in toxicity. We previously reported that the reincubation phase occurs with mitochondrial permeability transition (MPT) and increased oxidative stress (dichlorodihydrofluorescein fluorescence) (DCFH2). Since DCFH2 may be oxidized by multiple oxidative mechanisms, we investigated the role of reactive nitrogen species (RNS) leading to 3-nitrotyrosine in proteins by ELISA and by immunoblots. Incubation of APAP with hepatocytes for 2 h did not result in toxicity or protein nitration; however, washing hepatocytes and reincubating in media alone (2–5h) resulted in protein nitration which correlated with toxicity. Inclusion of the MPT inhibitor, cyclosporine A, in the reincubation media eliminated toxicity and protein nitration. The general nitric oxide synthase (NOS) inhibitor L-NMMA and the neuronal NOS (NOS1) inhibitor, 7-nitroindazole, added in the reincubation media decreased toxicity and protein nitration; however, neither the inducible NOS (NOS2) inhibitors L-NIL (N6-(1-iminoethyl)-l-lysine) nor SAIT (S-(2-aminoethyl)isothiourea) decreased protein nitration or toxicity. The RNS scavengers, N-acetylcysteine, and high concentrations of APAP, added in the reincubation phase decreased toxicity and protein nitration. 7-Nitroindazole and cyclosporine A inhibited the APAP-induced loss of mitochondrial membrane potential when added in the reincubation phase. The data indicate a role for RNS in APAP induced toxicity.
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DOI：
10.1080/15287390701551399
发表时间：
2007-01-01
影响因子：
2.6
作者：
Burke, Angela S.;Redeker, Kelly;Hinson, Jack A.
通讯作者：
Hinson, Jack A.
影响因子：
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作者：
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影响因子：
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影响因子：
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