Methyl Vinyl Ketone, a Toxic Ingredient in Cigarette Smoke Extract, Modifies Glutathione in Mouse Melanoma Cells

Horiyama, Shizuyo; Takahashi, Y.; Hatai, Mayuko; Honda, Chie; Suwa, Kiyoko; Ichikawa, Atsushi; Yoshikawa, Norishige; Nakamura, Kazuki; Kunitomo, Masaru; Date, Sachiko; Masujima, Tsutomu; Takayama, Mitsuo

doi:10.1248/cpb.c14-00109

Cited by 17 publications

(27 citation statements)

References 21 publications

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“…[19][20][21] These carbonyl compounds rapidly react with intracellular GSH and form their corresponding adducts, thereby depleting cellular GSH 4,22) and leading to cell damage. 4,22,23) In addition, it has been reported that a substantial amount of GSH in epithelial cells exposed to the gaseous phase of cigarette smoke is irreversibly modified to the GSH-ACR adduct (m/z 364) and GSH-CA adduct (m/z 378) by direct infusion mass spectra of the cell lysate, but the GSH-MVK adduct (m/z 378) and GSH-CA adduct are not separated. 7) Furthermore, it has been reported that the GSH-ACR and GSH-CA adducts can be detected in airway epithelial cells exposed to cigarette smoke, although the rela- tive yields were limited.…”

Section: Discussionmentioning

confidence: 99%

“…In a previous study, we revealed that MVK has a fairly stronger cytotoxicity than CA in mouse melanoma cells. 4) This phenomenon can be explained by the fact that aldo-keto reductases in the cells are more sensitive to the GSH-CA adduct than the GSH-MVK adduct, although further study is needed to elucidate such differences in the cytotoxicity of α,β-unsaturated carbonyls in cigarette smoke.…”

Section: )mentioning

confidence: 99%

“…The cytotoxic effects of cigarette smoke have been shown to be due to the presence of many oxidants and reactive aldehydes. [2][3][4] Among them, the α,β-unsaturated aldehydes acrolein (ACR) and crotonaldehyde (CA), abundantly present in cigarette smoke, have been often reported to strongly react with protein and peptide thiol groups. [5][6][7][8][9][10] In particular, glutathione (GSH), a tripeptide containing a thiol group, is an important antioxidant and essential cofactor for antioxidant enzyme glutathione peroxidase.…”

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confidence: 99%

“…GSH-CA and GSH-ACR adducts are barely detected, whereas the GSH-MVK adduct is clearly detected. 3,4) In this study, we chose B16-BL6 mouse melanoma cells, which are of a black and highly metastatic cell line, 15) in order to examine the effect of CSE and its active ingredients on the metastatic ability of these cancer cells.…”

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Intracellular Metabolism of α,β-Unsaturated Carbonyl Compounds, Acrolein, Crotonaldehyde and Methyl Vinyl Ketone, Active Toxicants in Cigarette Smoke: Participation of Glutathione Conjugation Ability and Aldehyde–Ketone Sensitive Reductase Activity

et al. 2016

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The major toxicants in cigarette smoke, α,β-unsaturated aldehydes, such as acrolein (ACR) and crotonaldehyde (CA), and α,β-unsaturated ketone, methyl vinyl ketone (MVK), are known to form Michael-type adducts with glutathione (GSH) and consequently cause intracellular GSH depletion, which is involved in cigarette smoke-induced cytotoxicity. We have previously clarified that exposure to cigarette smoke extract (CSE) of a mouse melanoma cell culture medium causes rapid reduction of intracellular GSH levels, and that the GSH-MVK adduct can be detected by LC/MS analysis while the GSH-CA adduct is hardly detected. In the present study, to clarify why the GSH-CA adduct is difficult to detect in the cell medium, we conducted detailed investigation of the structures of the reaction products of ACR, CA, MVK and CSE in the GSH solution or the cell culture medium. The mass spectra indicated that in the presence of the cells, the GSH-CA and GSH-ACR adducts were almost not detected while their corresponding alcohols were detected. On the other hand, both the GSH-MVK adducts and their reduced products were detected. In the absence of the cells, the reaction of GSH with all α,β-unsaturated carbonyls produced only their corresponding adducts. These results show that the GSH adducts of α,β-unsaturated aldehydes, CA and ACR, are quickly reduced by certain intracellular carbonyl reductase(s) and excreted from the cells, unlike the GSH adduct of α,β-unsaturated ketone, MVK. Such a difference in reactivity to the carbonyl reductase might be related to differences in the cytotoxicity of α,β-unsaturated aldehydes and ketones.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Intracellular Metabolism of α,β-Unsaturated Carbonyl Compounds, Acrolein, Crotonaldehyde and Methyl Vinyl Ketone, Active Toxicants in Cigarette Smoke: Participation of Glutathione Conjugation Ability and Aldehyde–Ketone Sensitive Reductase Activity

et al. 2016

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“…Glutathione adduct formation is known as a major cellular defense mechanism against acrolein and MVK . When we examined the levels of total and reduced glutathione (GSH), they were significantly decreased in Nox1 ‐disrupted clones (Fig.…”

Section: Resultsmentioning

confidence: 99%

NOX1/NADPH oxidase regulates the expression of multidrug resistance‐associated protein 1 and maintains intracellular glutathione levels

et al. 2019

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The involvement of superoxide‐generating NADPH oxidase (NOX) in the cytotoxic effects of cigarette smoke extracts has been documented. However, the underlying molecular mechanisms and NOX isoform involved have not been fully clarified. Among the different NADPH oxidase isoforms identified so far, NOX1 and NOX4 were found to be expressed in rat H9c2 cardiomyocytes. When H9c2 cells were exposed to acrolein or methyl vinyl ketone (MVK), major toxic components of cigarette smoke extracts, a dose‐dependent decline in cell viability was observed. Unexpectedly, disruption of Nox1 as well as Nox4 significantly exacerbated cytotoxicity induced by acrolein or MVK. Compared with Nox4‐disrupted cells, Nox1‐disrupted cells were more vulnerable to acrolein and MVK at lower concentrations. Disruption of Nox1 markedly attenuated the levels of total and reduced glutathione (GSH) in H9c2 clones. Reduction in the cystine level in the culture medium to deplete intracellular GSH significantly exacerbated acrolein or MVK‐induced cytotoxicity. Nox1 disruption neither attenuated the level of glutamate–cystine antiporter protein nor the activity of glutamate–cysteine ligase, both rate‐limiting factors for GSH synthesis. On the other hand, increased expression of multidrug resistance‐associated protein 1 (MRP1), which mediates glutathione efflux, was demonstrated in Nox1‐disrupted cells. The augmented toxicity of acrolein and MVK in these cells was partially but significantly blunted in the presence of an MRP1 inhibitor, reversan. Taken together, these results show that NOX1/NADPH oxidase regulates the expression of MRP1 to maintain intracellular GSH levels in cardiomyocytes and protect against cytotoxic components of cigarette smoke extracts. A novel crosstalk between NOX1 and MRP1 was demonstrated in this study.

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RNAi-mediated gene knockdown of progesterone 5β-reductases in Digitalis lanata reduces 5β-cardenolide content

et al. 2021

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Key message Studying RNAi-mediated DlP5βR1 and DlP5βR2 knockdown shoot culture lines of Digitalis lanata, we here provide direct evidence for the participation of PRISEs (progesterone 5β-reductase/iridoid synthase-like enzymes) in 5β-cardenolide formation. Abstract Progesterone 5β-reductases (P5βR) are assumed to catalyze the reduction of progesterone to 5β-pregnane-3,20-dione, which is a crucial step in the biosynthesis of the 5β-cardenolides. P5βRs are encoded by VEP1-like genes occurring ubiquitously in embryophytes. P5βRs are substrate-promiscuous enone-1,4-reductases recently termed PRISEs (progesterone 5β-reductase/iridoid synthase-like enzymes). Two PRISE genes, termed DlP5βR1 (AY585867.1) and DlP5βR2 (HM210089.1) were isolated from Digitalis lanata. To give experimental evidence for the participation of PRISEs in 5β-cardenolide formation, we here established several RNAi-mediated DlP5βR1 and DlP5βR2 knockdown shoot culture lines of D. lanata. Cardenolide contents were lower in D. lanata P5βR-RNAi lines than in wild-type shoots. We considered that the gene knockdowns may have had pleiotropic effects such as an increase in glutathione (GSH) which is known to inhibit cardenolide formation. GSH levels and expression of glutathione reductase (GR) were measured. Both were higher in the Dl P5βR-RNAi lines than in the wild-type shoots. Cardenolide biosynthesis was restored by buthionine sulfoximine (BSO) treatment in Dl P5βR2-RNAi lines but not in Dl P5βR1-RNAi lines. Since progesterone is a precursor of cardenolides but can also act as a reactive electrophile species (RES), we here discriminated between these by comparing the effects of progesterone and methyl vinyl ketone, a small RES but not a precursor of cardenolides. To the best of our knowledge, we here demonstrated for the first time that P5βR1 is involved in cardenolide formation. We also provide further evidence that PRISEs are also important for plants dealing with stress by detoxifying reactive electrophile species (RES).

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Methyl Vinyl Ketone, a Toxic Ingredient in Cigarette Smoke Extract, Modifies Glutathione in Mouse Melanoma Cells

Cited by 17 publications

References 21 publications

Intracellular Metabolism of α,β-Unsaturated Carbonyl Compounds, Acrolein, Crotonaldehyde and Methyl Vinyl Ketone, Active Toxicants in Cigarette Smoke: Participation of Glutathione Conjugation Ability and Aldehyde–Ketone Sensitive Reductase Activity

Intracellular Metabolism of α,β-Unsaturated Carbonyl Compounds, Acrolein, Crotonaldehyde and Methyl Vinyl Ketone, Active Toxicants in Cigarette Smoke: Participation of Glutathione Conjugation Ability and Aldehyde–Ketone Sensitive Reductase Activity

NOX1/NADPH oxidase regulates the expression of multidrug resistance‐associated protein 1 and maintains intracellular glutathione levels

RNAi-mediated gene knockdown of progesterone 5β-reductases in Digitalis lanata reduces 5β-cardenolide content

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