Differential effect of covalent protein modification and glutathione depletion on the transcriptional response of Nrf2 and NF-kappaB.
Differential effect of covalent protein modification and glutathione depletion on the transcriptional response of Nrf2 and NF-kappaB.
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DOI：
10.1016/j.bcp.2010.04.004
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发表时间：
2010-08-01
影响因子：
5.8
通讯作者：
Park BK
中科院分区：
文献类型：
--
作者：
Chia AJ;Goldring CE;Kitteringham NR;Wong SQ;Morgan P;Park BK
CRM激活NRF2，但抑制NF-κB，而没有共价修饰的GSH耗竭激活NRF2和NF-κB，这导致细胞保护免受氧化还原扰动的潜在有害影响。与暴露于进行肝脏代谢的治疗药物相关的肝损伤可能涉及反应性代谢物的形成。这些可能会引起氧化还原扰动，从而导致氧化应激以及蛋白质修饰，从而激活或抑制细胞转录反应。然而，这些挑战同时对多个转录途径的影响仍不清楚。我们研究了两个已知对亲电应激和氧化还原扰动敏感的转录因子，NRF2和NF-κB，在小鼠肝细胞中。用对乙酰氨基酚的反应性代谢物N-乙酰基-对苯喹亚胺、模型电泳剂二硝基氯苯和谷氨酸-半胱氨酸连接酶抑制剂丁硫氨酸-磺胺亚胺诱导细胞应激。NAPQI、DNCB和BSO都能引起谷胱甘肽(GSH)耗竭，但只有NAPQI和DNCB能与蛋白质共价结合。我们还利用RNA干扰来操纵Keap1(NRF2的抑制物)、NRF2本身和NF-κB-P65，以了解它们在药物应激反应中的作用。三种化学物质均可诱导NRF2，但只有在BSO处理后，NF-κB结合活性才增加。事实上，暴露于NAPQI和DNCB后，NF-κB结合活性降低。当KEAP1的RNAi耗尽导致DNCB暴露后的毒性降低时，NRF2和NF-κB的耗尽增加了毒性。有趣的是，Keap1耗尽引起的Nrf2增加可以通过NF-κB的共同耗尽而被逆转。我们证明了Keap1/Nrf2和NF-κB对共价结合蛋白质的亲电体和与谷胱甘肽耗竭相关的氧化还原扰动的反应不同，并且串扰可能使NF-κB能够在细胞应激过程中部分影响NRF2的表达。
CRMs activate Nrf2, but inhibit NF-κB, and GSH depletion without covalent modification activates both Nrf2 and NF-κB. This leads to cellular protection against the potentially harmful effects of redox perturbation. Liver injury associated with exposure to therapeutic agents that undergo hepatic metabolism can involve the formation of reactive metabolites. These may cause redox perturbation which can result in oxidative stress as well as protein modification leading to activation or inhibition of cellular transcriptional responses. Nevertheless, the effects of these challenges on more than one transcriptional pathway simultaneously remain unclear. We have investigated two transcription factors known to be sensitive to electrophilic stress and redox perturbation, Nrf2 and NF-κB, in mouse liver cells. Cellular stress was induced by the probes: N-acetyl-p-benzoquinineimine (NAPQI), the reactive metabolite of acetaminophen; dinitrochlorobenzene (DNCB), a model electrophile; and buthionine (S,R)-sulfoximine (BSO), an inhibitor of glutamate-cysteine ligase. NAPQI, DNCB and BSO can all cause glutathione (GSH) depletion; however only NAPQI and DNCB can covalently bind proteins. We also employed RNAi to manipulate Keap1 (the inhibitor of Nrf2), Nrf2 itself and NF-κB-p65, to understand their roles in the response to drug stress. All three chemicals induced Nrf2, but NF-κB binding activity was only increased after BSO treatment. In fact, NF-κB binding activity decreased after exposure to NAPQI and DNCB. While RNAi depletion of Keap1 led to reduced toxicity following exposure to DNCB, depletion of Nrf2 and NF-κB augmented toxicity. Interestingly, increased Nrf2 caused by Keap1 depletion was reversed by co-depletion of NF-κB. We demonstrate that Keap1/Nrf2 and NF-κB respond differently to electrophiles that bind proteins covalently and the redox perturbation associated with glutathione depletion, and that crosstalk may enable NF-κB to partly influence Nrf2 expression during cellular stress.
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影响因子：
5.3
作者：
Itoh, K;Mochizuki, M;Yamamoto, M
通讯作者：
Yamamoto, M
影响因子：
10.5
作者：
Itoh, K;Wakabayashi, N;Yamamoto, M
通讯作者：
Yamamoto, M
影响因子：
4.8
作者：
Cristofanon, Silvia;Morceau, Franck;Diederich, Marc
通讯作者：
Diederich, Marc
DOI：
10.1006/bbrc.1997.6943
发表时间：
1997-07-18
影响因子：
3.1
作者：
Itoh, K;Chiba, T;Nabeshima, Y
通讯作者：
Nabeshima, Y
DOI：
10.1152/ajpcell.2001.281.2.c719
发表时间：
2001-08-01
影响因子：
5.5
作者：
Huang, AN;Xiao, H;Keaney, JF
通讯作者：
Keaney, JF