Nrf2 protects against oxidative damage induced by hemoglobin in the liver of grass carp ( Ctenopharyngodon idella)

被引:1
|
作者
Li, Ningjing [1 ]
Song, Jialing [1 ]
Yang, Yan [1 ]
Huang, Xiaoman [1 ]
Tian, Ye [1 ]
Chen, Bing [2 ]
Lin, Li [1 ]
Qin, Zhendong [1 ]
机构
[1] Zhongkai Univ Agr & Engn, Coll Anim Sci & Technol, Guangzhou 510225, Guangdong, Peoples R China
[2] Guangdong Acad Agr Sci, Inst Anim Sci, Lab Anim Nutr & Feed Sci South China, Guangdong Key Lab Anim Breeding & Nutr,Minist Agr, Guangzhou 510640, Peoples R China
基金
中国国家自然科学基金;
关键词
Nrf2; AMPK; Hb; Oxidative damage; Ctenopharyngodon idella; TISSUE-DAMAGE; HEMOLYSIS; STRESS; HEME; HOMEOSTASIS; IRON; AMPK;
D O I
10.1016/j.bbadis.2024.167600
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Hemoglobin (Hb) releases during hemorrhaging and causes oxidative damage, further exacerbates the development of multiple diseases. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that regulates cellular defenses against toxic and oxidative challenges. However, the regulation mechanism of Nrf2 in Hb-induced oxidative stress remains unclear in teleost. To accomplish this goal, a hemolysis model was established by injecting grass carp with phenylalanine (PHZ), and the immunofluorescence analysis (IFA) and hematoxylin and eosin (H&E) staining revealed that PHZ-induced hemolysis caused Hb accumulation and hepatic vacuolization, resulted in tissue damage. Prussian blue, Sirius red, and Masson staining results revealed significant iron deposition and extensive collagen fiber accumulation in the liver. IFA and immunohistochemical analyses demonstrated that PHZ-induced hemolysis markedly increased the production of reactive oxygen species (ROS), malondialdehyde (MDA), and 4-hydroxynonenal (4-HNE). The quantitative real-time PCR (qRT-PCR) analysis data revealed that the PHZ-induced hemolysis also significantly upregulated the expression of antioxidant-related genes through activation of the adenosine 5 '-monophosphate (AMP)-activated protein kinase (AMPK)/Nrf2 signaling pathway. To further explore the molecule regulation mechanism of PHZ-induced hemolysis, the RNA-seq analysis was performed, and the data revealed that the AMPK/Nrf2 and multiple programmed cell death pathways, including ferroptosis, autophagy, apoptosis, and necroptosis in PHZ injection groups were significant upregulated. In vitro, the hemin supplementation activated the expression of target genes in the AMPK/Nrf2 pathway detected by qRT-PCR. To further verify the regulation function of Nrf2, an Nrf2 activator (4OI) was supplemented, and the flow cytometer analysis results suggested that the Hb-induced cell damage was significantly attenuated. However, the supplementary of ML385 down-regulated the AMPK/Nrf2 pathway and aggravated the hemin induced cell death. In conclusion, these findings highlight the critical regulatory role of the AMPK/Nrf2 signaling pathway in protecting against Hb-induced oxidative damage in the liver of grass carp.
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页数:15
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