Ablation of aldehyde reductase aggravates carbon tetrachloride-induced acute hepatic injury involving oxidative stress and endoplasmic reticulum stress

Akihara, Ryusuke; Homma, Takujiro; Lee, Jae Yong; Yamada, Ken Ichi; Miyata, Satoshi; Fujii, Junichi

doi:10.1016/j.bbrc.2016.08.022

Cited by 14 publications

(10 citation statements)

References 38 publications

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“…It is involved in the reduction of aldehydes to corresponding alcohols 44,45 as well as the reduction of D-glucuronic acid in the biosynthesis of ascorbic acid. 46,47 Gpx1, the second most stable gene in long-term cultivation, is coding for glutathione peroxidase 1 and is responsible for the balance of intracellular redox environment. 48 Neither of these two genes had been reported to use as reference genes in RT-qPCR analyses before.…”

Section: Discussionmentioning

confidence: 99%

Validation and identification of reference genes in Chinese hamster ovary cells for Fc-fusion protein production

Zhang

et al. 2020

Exp Biol Med (Maywood)

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Chinese hamster ovary cells are the predominant cell lines used for bio-therapeutic production. Real-time quantitative PCR (RT-qPCR) and transcriptomics are powerful tools to understand and optimize the Chinese hamster ovary cells for higher productivity or better control of product qualities. Reliable reference genes, which were proved to be experiment-specific, are critical yardsticks. In this study, we compared expression stability of 20 candidate reference genes at mRNA level, including commonly used housekeeping genes, previous literature reported genes in Chinese hamster ovary cells producing an intact antibody, and new candidates suggested by our RNA-seq transcriptomic database, in RT-qPCR reactions in Fc-fusion protein-producing Chinese hamster ovary cells with various productivity during long-term cultivation and fed-batch cultures at 26 different conditions. geNorm, NormFinder, BestKeeper, and ΔCt programs and methods were utilized to analyze the gene expression stability and gave an overall ranking. Akr1a1, Gpx1, and Aprt in long-term cultivation and Akr1a1, Rps16 in fed-batch culture, which have not been reported previously, exhibited the highest stability of gene expression, while Pabpn1, Hirip3, and Actb in both sets of experiments together with Atp5f1 in long-term passage process showed the weakest stability. The results were then validated using GLP1-Fc (Glucagon-like peptide-1 Fc fusion protein) gene as the target with determined expression level which were doubly confirmed by both absolute RT-qPCR and confocal microscopy. These new references should be considered for the investigations on Chinese hamster ovary cells in related research. Impact statement In order to reveal potential genotype-phenotype relationship, RT-qPCR reactions are frequently applied which require validated and reliable reference genes. With the investigation on long-term passage and fed-batch cultivation of CHO cells producing an Fc-fusion protein, four new reference genes–Akr1a1, Gpx1, Aprt, and Rps16, were identified from 20 candidates with the aid of geNorm, NormFinder, BestKeeper, and ΔCt programs and methods. This article provided more verified options in reference gene selection in related research on CHO cells.

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Section: Discussionmentioning

confidence: 99%

Validation and identification of reference genes in Chinese hamster ovary cells for Fc-fusion protein production

Zhang

et al. 2020

Exp Biol Med (Maywood)

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“…The livers of AKR1A −/− mice are significantly heavier than those from AKR1A +/+ mice [ 185 ], which may be associated with altered glycogen metabolism [ 188 ] and differential responses to hepatotoxic compounds in AKR1A −/− mice [ 189 , 190 ]. Treatment with CCl 4 causes severe hepatic damage with enhanced steatosis in the livers of AKR1A −/− mice compared to WT mice, and Asc supplementation reverses these changes, making them comparable to those in WT mice [ 189 ]. The aggravation in hepatic damage is consistent with observations regarding RGN −/− mice [ 158 ].…”

Section: In Vivo Antioxidative Action Of Asc Revealed By Animal Studiesmentioning

confidence: 99%

“…The aggravation in hepatic damage is consistent with observations regarding RGN −/− mice [ 158 ]. However, there are contradictions in the actions of supplemented Asc between AKR1A −/− mice and RGN −/− mice; while Asc is protective for AKR1A −/− mice [ 189 ], it is detrimental for RGN −/− mice [ 158 ]. These seemingly inconsistent observations might be due to differences in the unique actions of these genes, so clarification is also awaited regarding this issue as well.…”

Section: In Vivo Antioxidative Action Of Asc Revealed By Animal Studiesmentioning

confidence: 99%

Ascorbate Is a Primary Antioxidant in Mammals

Fujii

Osaki

2022

Molecules

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Ascorbate (vitamin C in primates) functions as a cofactor for a number of enzymatic reactions represented by prolyl hydroxylases and as an antioxidant due to its ability to donate electrons, which is mostly accomplished through non-enzymatic reaction in mammals. Ascorbate directly reacts with radical species and is converted to ascorbyl radical followed by dehydroascorbate. Ambiguities in physiological relevance of ascorbate observed during in vivo situations could be attributed in part to presence of other redox systems and the pro-oxidant properties of ascorbate. Most mammals are able to synthesize ascorbate from glucose, which is also considered to be an obstacle to verify its action. In addition to animals with natural deficiency in the ascorbate synthesis, such as guinea pigs and ODS rats, three strains of mice with genetic removal of the responsive genes (GULO, RGN, or AKR1A) for the ascorbate synthesis have been established and are being used to investigate the physiological roles of ascorbate. Studies using these mice, along with ascorbate transporter (SVCT)-deficient mice, largely support its ability in protection against oxidative insults. While combined actions of ascorbate in regulating epigenetics and antioxidation appear to effectively prevent cancer development, pharmacological doses of ascorbate and dehydroascorbate may exert tumoricidal activity through redox-dependent mechanisms.

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“…The copyright holder for this preprint this version posted December 11, 2022. ; https://doi.org/10.1101/2022.12.07.519420 doi: bioRxiv preprint AKR1A1 levels are abundant in the liver and can prevent carbon tetrachloride- [10],…”

Section: Introductionmentioning

confidence: 99%

Aldo-Keto Reductase Family 1 Member A1 (AKR1A1) Deficiency Exacerbates Alcohol-Induced Hepatic Oxidative Stress, Inflammation, Steatosis, and Fibrosis

Lan

Chen

Yen

et al. 2022

Preprint

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Background: Alcohol-associated liver disease (ALD) covers a wide range of hepatic lesions that depend on the amount and duration of alcohol consumption, from early and reversible conditions to hepatic steatosis and severe lesions, including steatohepatitis and alcoholic fibrosis, to irreversible cirrhosis. AKR1A1, an aldo-keto reductase family member, participates in the detoxification of alcohol-derived acetaldehyde, but its role in ALD remains unclear. In this study, we studied the role of AKR1A1 in the development of ALD using Akr1a1-/- knockout mice and palmitic acid/oleic acid (P/O) plus ethanol-treated AML12 hepatocyte cells. Methods: Levels of AKR1A1 were measured in mice fed with the Lieber-DeCarli diet containing 5% alcohol (alcohol-fed, AF) or control liquid diet (pair-fed, PF). The effects of AKR1A1 on the liver function, inflammation, oxidative stress, lipid accumulation, and fibrosis were assessed in AF-induced Akr1a1-/- and ICR control mice. Results: Data showed that AF-Akr1a1-/- mice exhibited an exacerbation of liver injury and increased gene and protein levels of inflammatory mediators, oxidative stress, lipid accumulation, and fibrosis, whilst decreased expression of antioxidant enzymes in their livers than the AF-ICR mice. Therefore, loss of AKR1A1 can activate 4-HNE/p53 signaling to modulate ROS and antioxidant balance, increase lipid peroxidation, fatty acid synthesis and lipid droplet formation, reduced fatty acid β-oxidation, and elevated proinflammatory and fibrotic mediator, eventually exacerbate the ALD. In in vitro study, we further demonstrated that knockdown of Akr1a1 aggravated the effects of alcohol plus P/O-induced oxidative stress and steatosis, LPS-stimulated inflammation, and TGF-β1-induced fibrosis in AML12 hepatocyte cells. Conclusion: our results revealed that AKR1A1 exerts protective effects on alcohol-induced liver injury, steatosis, and fibrosis, possibly by regulating the 4-HNE-p53 signaling pathway.

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Ablation of aldehyde reductase aggravates carbon tetrachloride-induced acute hepatic injury involving oxidative stress and endoplasmic reticulum stress

Cited by 14 publications

References 38 publications

Validation and identification of reference genes in Chinese hamster ovary cells for Fc-fusion protein production

Validation and identification of reference genes in Chinese hamster ovary cells for Fc-fusion protein production

Ascorbate Is a Primary Antioxidant in Mammals

Aldo-Keto Reductase Family 1 Member A1 (AKR1A1) Deficiency Exacerbates Alcohol-Induced Hepatic Oxidative Stress, Inflammation, Steatosis, and Fibrosis

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