Nutr Res Pract.  2022 Dec;16(6):729-744. 10.4162/nrp.2022.16.6.729.

L-Methionine inhibits 4-hydroxy-2-nonenal accumulation and suppresses inflammation in growing rats

• Wang Z
• Liang M
• Li H
• Liu B
• Yang L

• KMID: 2536507
• DOI: http://doi.org/10.4162/nrp.2022.16.6.729

Abstract

BACKGROUND/OBJECTIVES
4-Hydroxy-2-nonenal (HNE) is a biomarker for oxidative stress to induce inflammation. Methionine is an essential sulfur-containing amino acid with antioxidative activity. On the other hand, the evidence on whether and how methionine can depress HNE-derived inflammation is lacking. In particular, the link between the regulation of the nuclear factor-κB (NF-κB) signaling pathway and methionine intake is unclear. This study examined the link between depression from HNE accumulation and the antiinflammatory function of L-methionine in rats.
MATERIALS/METHODS
Male Wistar rats (3-week-old, weighing 70–80 g) were administered different levels of L-methionine orally at 215.0, 268.8, 322.5, and 430.0 mg/kg body weight for two weeks. The control group was fed commercial pellets. The hepatic HNE contents and the protein expression and mRNA levels of the inflammatory mediators were measured. The interleukin-10 (IL-10) and glutathione S-transferase (GST) levels were also estimated.
RESULTS
Compared to the control group, hepatic HNE levels were reduced significantly in all groups fed L-methionine, which were attributed to the stimulation of GST by L-methionine. With decreasing HNE levels, L-methionine inhibited the activation of NF-κB by up-regulating inhibitory κBα and depressing phosphoinositide 3 kinase/protein kinase B. The mRNA levels of the inflammatory mediators (cyclooxygenase-2, interleukin-1β, interleukin-6, inducible nitric oxide synthase, tumor necrotic factor alpha) were decreased significantly by L-methionine. In contrast, the protein expression of these inflammatory mediators was effectively down regulated by L-methionine. The anti-inflammatory action of L-methionine was also reflected by the up-regulation of IL-10.
CONCLUSIONS
This study revealed a link between the inhibition of HNE accumulation and the depression of inflammation in growing rats, which was attributed to L-methionine availability. The anti-inflammatory mechanism exerted by L-methionine was to inhibit NF-κB activation and to up-regulate GST.

Keyword

L-Methionine; 4-hydroxy-2-nonenal; inflammation; NF-κB; GST

Figure

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  Fig. 1 Schematic procedure of the animal experiment.

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  Fig. 2 Hepatic contents of HNE in growing rats. Hepatic contents of HNE in growing rats after the oral administration of L-methionine for 14 days. The values are expressed as the means ± SEM (n = 6).HNE, 4-hydroxy-2-nonenal; C, control; M50, basal diet and administered L-methionine orally at 215.0 mg/kg bw; M62.5, basal diet and administered L-methionine orally at 268.8 mg/kg bw; M75, basal diet and administered L-methionine orally at 322.5 mg/kg bw; M100, basal diet and administered L-methionine orally at 430.0 mg/kg bw.*P < 0.05, in comparison with C.

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  Fig. 3 Hepatic protein expression and mRNA level of COX-2 after the oral administration of L-methionine for 14 days. (A) Hepatic protein expression of COX-2. (B) Hepatic mRNA level of COX-2. The values are expressed as the means ± SEM (n = 6).COX-2, cyclooxygenase-2; C, control; M50, basal diet and administered L-methionine orally at 215.0 mg/kg bw; M62.5, basal diet and administered L-methionine orally at 268.8 mg/kg bw; M75, basal diet and administered L-methionine orally at 322.5 mg/kg bw; M100, basal diet and administered L-methionine orally at 430.0 mg/kg bw.*P < 0.05, in comparison with C.

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  Fig. 4 Hepatic protein expression and mRNA levels of inflammatory mediators after the oral administration of L-methionine for 14 days. (A) Hepatic protein expression and mRNA levels of IL-1β. (B) Hepatic protein expressions and mRNA levels of IL-6. (C) Hepatic protein expressions and mRNA levels of iNOS. (D) Hepatic protein expressions and mRNA levels of TNF-α. Values are the means ± SEM (n = 6).IL-1β, interleukin-1β; IL-6, interleukin-6; iNOS, inducible nitric oxide synthase; TNF-α, tumor necrotic factor-alpha; C, control; M50, basal diet and administered L-methionine orally at 215.0 mg/kg bw; M62.5, basal diet and administered L-methionine orally at 268.8 mg/kg bw; M75, basal diet and administered L-methionine orally at 322.5 mg/kg bw; M100, basal diet and administered L-methionine orally at 430.0 mg/kg bw.*P < 0.05, compared with C.

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  Fig. 5 Hepatic protein expressions and mRNA levels of NF-κB1 and RelA after the oral administration of L-methionine for 14 days. (A) Hepatic protein expressions and mRNA levels of NF-κB1. (B) Hepatic protein expressions and mRNA levels of RelA. The values are expressed as the means ± SEM (n = 6).NF-κB1, nuclear factor-κB1; RelA, reticuloendotheliosis viral oncogene homolog A; C, control; M50, basal diet and administered L-methionine orally at 215.0 mg/kg bw; M62.5, basal diet and administered L-methionine orally at 268.8 mg/kg bw; M75, basal diet and administered L-methionine orally at 322.5 mg/kg bw; M100, basal diet and administered L-methionine orally at 430.0 mg/kg bw.*P < 0.05, compared with C.

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  Fig. 6 Effects of L-methionine on nuclear factor-κB activation after the oral administration for 14 days. (A) Hepatic protein expressions of IκBα. (B) Hepatic mRNA levels of IκBα. (C) Nuclear and cytosolic protein contents of p50. (D) Nuclear and cytosolic protein contents of p65. The values are expressed as the means ± SEM (n = 6).IκBα, inhibitory κBα; C, control; M50, basal diet and administered L-methionine orally at 215.0 mg/kg bw; M62.5, basal diet and administered L-methionine orally at 268.8 mg/kg bw; M75, basal diet and administered L-methionine orally at 322.5 mg/kg bw; M100, basal diet and administered L-methionine orally at 430.0 mg/kg bw.*P < 0.05, in comparison with C.

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  Fig. 7 Hepatic protein expressions and mRNA levels of PI3K and AKT after the oral administration of L-methionine for 14 days. (A) Hepatic protein expressions and mRNA levels of PI3K. (B) Hepatic protein expressions and mRNA levels of AKT. The values are expressed as the means ± SEM (n = 6).PI3K, phosphoinositide 3 kinase; AKT, protein kinase B; C, control; M50, basal diet and administered L-methionine orally at 215.0 mg/kg bw; M62.5, basal diet and administered L-methionine orally at 268.8 mg/kg bw; M75, basal diet and administered L-methionine orally at 322.5 mg/kg bw; M100, basal diet and administered L-methionine orally at 430.0 mg/kg bw.*P < 0.05, compared with C.

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  Fig. 8 Hepatic protein expression and mRNA levels of IL-10 and GST in growing rats. (A) Hepatic protein expressions of IL-10. (B) Hepatic mRNA levels of IL-10. (C) Hepatic protein expressions of GST. (D) Hepatic mRNA levels of GST. The values are expressed as the means ± SEM (n = 6).IL-10, interleukin-10; GST, glutathione S-transferase α1; C, control; M50, basal diet and administered L-methionine orally at 215.0 mg/kg bw; M62.5, basal diet and administered L-methionine orally at 268.8 mg/kg bw; M75, basal diet and administered L-methionine orally at 322.5 mg/kg bw; M100, basal diet and administered L-methionine orally at 430.0 mg/kg bw.*P < 0.05, compared with C.

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  Fig. 9 L-Methionine inhibits HNE accumulation and depresses inflammation by inhibiting NF-κB activation and up-regulating GST.Nrf2, nuclear factor erythroid 2-related factor 2; GST, glutathione S-transferase; PI3K, phosphoinositide 3 kinase; AKT, protein kinase B; HNE, 4-hydroxy-2-nonenal; IκBα, inhibitory κBα; NF-κB, nuclear factor-κB; COX-2, cyclooxygenase-2; TNF-α, tumor necrotic factor alpha; IL, interleukin; iNOS, inducible nitric oxide synthase.

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