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. 2024 Aug 19;37(8):1306-1314.
doi: 10.1021/acs.chemrestox.4c00052. Epub 2024 Jul 27.

Quantitative Analysis of Glutathione and Carnosine Adducts with 4-Hydroxy-2-nonenal in Muscle in a hSOD1G93A ALS Rat Model

Pablo V M Reis  1 , Bianca S Vargas  1 , Rafael A Rebelo  1 , Mariana P Massafera  1 , Fernanda M Prado  1 , Hector Oreliana  1 , Henrique V de Oliveira  1 , Florêncio P Freitas  1 , Graziella E Ronsein  1 , Sayuri Miyamoto  1 , Paolo Di Mascio  1 , Marisa H G Medeiros  1
Affiliations

Quantitative Analysis of Glutathione and Carnosine Adducts with 4-Hydroxy-2-nonenal in Muscle in a hSOD1G93A ALS Rat Model

Pablo V M Reis et al. Chem Res Toxicol. .

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the dysfunction and death of motor neurons through multifactorial mechanisms that remain unclear. ALS has been recognized as a multisystemic disease, and the potential role of skeletal muscle in disease progression has been investigated. Reactive aldehydes formed as secondary lipid peroxidation products in the redox processes react with biomolecules, such as DNA, proteins, and amino acids, resulting in cytotoxic effects. 4-Hydroxy-2-nonenal (HNE) levels are elevated in the spinal cord motor neurons of ALS patients, and HNE-modified proteins have been identified in the spinal cord tissue of an ALS transgenic mice model, suggesting that reactive aldehydes can contribute to motor neuron degeneration in ALS. One biological pathway of aldehyde detoxification involves conjugation with glutathione (GSH) or carnosine (Car). Here, the detection and quantification of Car, GSH, GSSG (glutathione disulfide), and the corresponding adducts with HNE, Car-HNE, and GS-HNE, were performed in muscle and liver tissues of a hSOD1G93A ALS rat model by reverse-phase high-performance liquid chromatography coupled to electrospray ion trap tandem mass spectrometry in the selected reaction monitoring mode. A significant increase in the levels of GS-HNE and Car-HNE was observed in the muscle tissue of the end-stage ALS animals. Therefore, analyzing variations in the levels of these adducts in ALS animal tissue is crucial from a toxicological perspective and can contribute to the development of new therapeutic strategies.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Scheme 1
Scheme 1. Detoxification Products of trans-4-Hydroxynonenal Reaction with Either Glutathione or Carnosine
Figure 1
Figure 1
Representative chromatograms of the GS-HNE adducts in the liver of a control rat (A). The endogenous adduct (top) exhibits two equally intense peaks, representing biologically formed isomers. The labeled internal standard (lower) predominantly forms the first peak, indicating a difference in isomeric formation between in vitro and in vivo systems. GS-HNE levels in liver (B) and muscle tissue (C) of 70 and 120-day-old ALS rats along with the corresponding controls. Full MS2 of the GS-HNE adduct, showing the two fragments monitored in SRM mode and m/z 179.2 (D).
Figure 2
Figure 2
Representative chromatograms of GSSG and the GS-NEM adduct in the liver of a control rat (A). The reduced form of glutathione forms two equally intense peaks, indicating biological isomers. Endogenous (upper) and internal standard (lower) peaks for GSSG show the same profile, indicating that isomers formed both in vivo and in vitro. GSSG (B) and GS-NEM (C) levels in the liver. GSSG (D) and GS-NEM (E) levels in the muscle tissue of 70 and 120-day-old ALS rats along with the corresponding controls.
Figure 3
Figure 3
Representative chromatogram for control rat muscle sample, showing successful Carnosine and CAR-HNE separation, in both endogenous analytes (upper) and internal standards (lower) (A). Car-HNE shows two peaks in both in vivo and in vitro synthesis, indicating that endogenous (upper) and internal standard (lower) forms are composed of the same isomers. Levels of Carnosine (B) and Car-HNE (C) in muscle tissue of 70 and 120-day-old ALS rats along with corresponding controls. Full MS2 of the Car-HNE adduct (D).
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References

    1. Hardiman O.; Al-chalabi A.; Chio A.; Corr E. M.; Logroscino G.; Robberecht W.; Shaw P. J.; Simmons Z.; van den Berg L. H. Amyotrophic Lateral Sclerosis. Nat. Rev. Dis. Primers 2017, 3, 71.10.1038/nrdp.2017.71. - DOI
    1. Ingre C.; Roos P. M.; Piehl F.; Kamel F.; Fang F. Risk Factors for Amyotrophic Lateral Sclerosis. Clin. Epidemiol. 2015, 7, 181–193. 10.2147/CLEP.S37505. - DOI - PMC - PubMed
    1. Rosen D. R.; Siddique T.; Patterson D.; Figlewicz D. A.; Sapp P. II; Hentati A.; Donaldson D.; Goto J.; O’Regan J. P.; Deng H.; Rahmani Z.; Krizus A.; Mckenna-yasek D.; Cayabyab A.; Gaston S. M.; Berger R.; Tanzi R. E.; Halperin J. J.; Herzfeldt B.; Van den Bergh R.; Hung W.; Bird T.; Deng G.; Mulder D. W.; Smyth C.; Laing N. G.; Soriano E.; Pericak-Vance M. A.; Haines J.; Rouleau G. A.; Gusella J. S.; Horvitz H. R.; Brown Jr R. H. Mutations in Cu/Zn Superoxide Dismutase Gene Are Associated with Familial Amyptrophic Lateral Sclerosis. Nature 1993, 362, 59–62. 10.1038/362059a0. - DOI - PubMed
    1. Borchelt D. R.; Lee M. K.; Slunt H. S.; Guarnieri M.; Xu Z.; Wong P. C.; Brown R. H.; Price D. L.; Sisodia S. S.; Cleveland D. O. N. W. Superoxide Dismutase 1 with Mutations Linked to Familial Amyotrophic Lateral Sclerosis Possesses Significant Activity. Proc. Natl. Acad. Sci. U. S. A. 1994, 91, 8292–8296. 10.1073/pnas.91.17.8292. - DOI - PMC - PubMed
    1. Rabizadeh S.; Gralla E. B.; Borchelt D. R.; Gwinn R.; Valentine J. S.; Sisodia S.; Wong P.; Lee M.; Hahn H.; Bredesen D. E. Mutations Associated with Amyotrophic Lateral Sclerosis Convert Superoxide Dismutase from an Antiapoptotic Gene to a Proapoptotic Gene: Studies in Yeast and Neural Cells. Proc. Natl. Acad. Sci. U. S. A. 1995, 92, 3024–3028. 10.1073/pnas.92.7.3024. - DOI - PMC - PubMed