Acrolein Causes Inhibitor κB-independent Decreases in Nuclear Factor κB Activation in Human Lung Adenocarcinoma (A549) Cells

Horton, Noel D.; Biswal, Shyam; Corrigan, Lucindra L.; Bratta, Julie; Kehrer, James P.

doi:10.1074/jbc.274.14.9200

Cited by 88 publications

(68 citation statements)

References 49 publications

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“…Further studies demonstrated that both the basal and activated NF-B DNA binding activity was inhibited by acrolein due to a decrease in IB␣ phosphorylation and degradation (14). In contrast, inhibition of NF-B DNA binding activity in human lung adenocarcinoma cells was independent of IB␣ (15). In primary or immortalized human bronchial epithelial cells, acrolein (25 M) suppressed both IL-8 protein and mRNA levels reportedly through inactivation of NF-B and reduced IB␣ degradation (16).…”

Section: Discussionmentioning

confidence: 96%

Acrolein Inhibits Cytokine Gene Expression by Alkylating Cysteine and Arginine Residues in the NF-κB1 DNA Binding Domain

Lambert¹,

Li²,

Jonscher

et al. 2007

Journal of Biological Chemistry

107

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Cigarette smoke is a potent inhibitor of pulmonary T cell responses, resulting in decreased immune surveillance and an increased incidence of respiratory tract infections. The ␣,␤-unsaturated aldehydes in cigarette smoke (acrolein and crotonaldehyde) inhibited production of interleukin-2 (IL-2), IL-10, granulocyte-macrophage colony-stimulating factor, interferon-␥, and tumor necrosis factor-␣ by human T cells but did not inhibit production of IL-8. The saturated aldehydes (acetaldehyde, propionaldehyde, and butyraldehyde) in cigarette smoke were inactive. Acrolein inhibited induction of NF-B DNA binding activity after mitogenic stimulation of T cells but had no effect on induction of NFAT or AP-1. Acrolein inhibited NF-B1 (p50) binding to the IL-2 promoter in a chromatin immunoprecipitation assay by >99%. Using purified recombinant p50 in an electrophoretic mobility shift assay, we demonstrated that acrolein was 2000-fold more potent than crotonaldehyde in blocking DNA binding to an NF-B consensus sequence. Matrix-assisted laser desorption/ionization time-offlight and tandem mass spectrometry demonstrated that acrolein alkylated two amino acids (Cys-61 and Arg-307) in the DNA binding domain. Crotonaldehyde reacted with Cys-61, but not Arg-307, whereas the saturated aldehydes in cigarette smoke did not react with p50. These experiments demonstrate that aldehydes in cigarette smoke can regulate gene expression by direct modification of a transcription factor.Cigarette smoke produces profound suppression of pulmonary immunity, resulting in an increased incidence and severity of respiratory tract infections. A recent Institute of Medicine study concluded that smoking increased the incidence of influenza and bacterial pneumonia and accounted for 19,000 smoking-related deaths per year (1). Children infected with Mycobacterium tuberculosis are five times more likely to develop pulmonary tuberculosis if exposed to cigarette smoke (2), and smoking doubles the risk of developing Pneumocystis carinii pneumonia in human immunodeficiency virus-infected individuals (3). Several studies have demonstrated that smoking suppresses T and B cell responses in the lungs without affecting cells in the peripheral blood (4 -7), but little research has been done to elucidate the underlying mechanism behind this phenomenon.We have recently identified two classes of immunosuppressive compounds in cigarette smoke. The dihydroxyphenols (hydroquinone and catechol) in the particulate phase inhibit T cell proliferation by blocking cell cycle progression in late G 1 and S phase (8 -12). In addition, the ␣,␤-unsaturated aldehydes, acrolein (CH 2 ACHCHO) and crotonaldehyde (CH 3 CHACHCHO) in the gas phase of cigarette smoke inhibit the production of several proinflammatory cytokines including IL-2, 4 tumor necrosis factor-␣, and granulocyte-macrophage colony-stimulating factor, with an IC 50 of 3 and 6 M, respectively (13). The saturated aldehydes (acetaldehyde, propionaldehyde, and butyraldehyde) have IC 50 values of Ͼ1500 M. The typical cigarett...

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Section: Discussionmentioning

confidence: 96%

Acrolein Inhibits Cytokine Gene Expression by Alkylating Cysteine and Arginine Residues in the NF-κB1 DNA Binding Domain

Lambert¹,

Li²,

Jonscher

et al. 2007

Journal of Biological Chemistry

107

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“…Evaluation of this possibility would require the development of new analytical tools to detect acrolein-linked macromolecules in biological systems, perhaps via similar approaches to those used to identify 4-hydroxynonenal-linked lysine residues in adducted proteins (Tsai et al, 1998;Xu et al, 2000). Such methodologies would facilitate study of such questions as whether cross-linking contributes to the molecular effects of acrolein in exposed cells, including protein kinase C activation (Maddox et al, 2003) or alterations in the activity of various transcription factors (e.g., activator protein-1, nuclear factor-␤, or nuclear factor-E2-related factor-2) (Horton et al, 1999;Biswal et al, 2002;Tirumalai et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

Strong Protein Adduct Trapping Accompanies Abolition of Acrolein-Mediated Hepatotoxicity by Hydralazine in Mice

Kaminskas

Pyke²,

Burcham³

2004

The Journal of Pharmacology and Experimental Therapeutics

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Acrolein is a highly reactive ␣,␤-unsaturated aldehyde that readily alkylates nucleophilic centers in cell macromolecules. Typically, such reactions proceed via Michael addition chemistry, forming adducts that retain an electrophilic carbonyl group. Since these species participate in secondary deleterious reactions, we hypothesize that inactivation of carbonyl adducts may attenuate acrolein toxicity. Indeed, we recently established that the nucleophilic antihypertensive drug hydralazine readily "traps" acrolein adducts in cell proteins and strongly suppresses acrolein-mediated toxicity in isolated hepatocytes. This work sought to determine whether hydralazine prevents the in vivo hepatotoxicity of the acrolein precursor allyl alcohol in whole mice and whether adduct trapping accompanies any such hepatoprotection. Mice received allyl alcohol alone or in conjunction with several doses of hydralazine. Four hours later, mice were sacrificed to allow for the determination of liver enzymes in plasma as markers of hepatic injury, whereas livers were assessed for glutathione and hydralazine-stabilized protein adducts. Hydralazine afforded strong, dose-dependent protection against the increases in plasma marker enzymes but not the hepatic glutathione depletion produced by allyl alcohol. Western blotting revealed intense, dose-dependent adduct trapping by hydralazine in numerous liver proteins over a broad 26-to 200-kDA mass range. In keeping with these findings, immunohistochemical analysis of liver slices indicated diffuse, extranuclear adduct trapping by hydralazine that was uniformly distributed across the liver lobule, with partial localization in parenchymal cell membranes. These findings concur with our hypothesis that hydralazine readily inactivates reactive carbonyl-retaining protein adducts formed by acrolein, thereby preventing secondary reactions that trigger cellular death.

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“…For the experiments involving direct addition of acrolein to cell media ( Fig. 1), Krebs-Henseleit buffer, pH 7.3, supplemented with glucose (5 g/l) and pyruvate (1 mM) was used in place of RPMI 1640 medium, in an effort to minimize extracellular side-reactions between free acrolein and nucleophilic buffer constituents (Horton et al, 1999). RPMI 1640 medium was used in all experiments that involved the use of allyl alcohol as an intracellular acrolein precursor.…”

Section: Methodsmentioning

confidence: 99%

“…This reactivity underlies most of the cellular effects of acrolein, including alterations in the activity of transcription factors such as AP-1, nuclear factor B, and Nrf2 (Horton et al, 1999;Biswal et al, 2002;Tirumalai et al, 2002); inhibition of cytokine production (Li et al, 1997); and cell death (Li et al, 1997;Kern and Kehrer, 2002). Typically, cellular nucleophiles target acrolein's ␤-carbon, generating carbonyl-retaining Michael adducts (Esterbauer et al, 1991;Uchida et al, 1998a;Burcham and Fontaine, 2001).…”

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confidence: 99%

Protein Adduct-Trapping by Hydrazinophthalazine Drugs: Mechanisms of Cytoprotection Against Acrolein-Mediated Toxicity

Burcham¹,

Fontaine²,

Kaminskas³

et al. 2004

Molecular Pharmacology

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Acrolein is a highly toxic aldehyde involved in a number of diseases as well as drug-induced toxicities. Its pronounced toxicity reflects the readiness with which it forms adducts in proteins and DNA. As a bifunctional electrophile, initial reactions between acrolein and protein generate adducts containing an electrophilic center that can participate in secondary deleterious reactions (e.g., cross-linking). We hypothesize that inactivation of these reactive protein adducts with nucleophilic drugs may counteract acrolein toxicity. Because we previously observed that 1-hydrazinophthalazine (hydralazine) strongly diminishes the toxicity of the acrolein precursor allyl alcohol, we explored the possibility that hydralazine targets reactive acrolein adducts in proteins. We report that hydralazine abolished the immunoreactivity of an acrolein-modified model protein (bovine serum albumin), but only if the drug was added to the protein within 30 min of commencing modification by acrolein. The ability of a range of carbonyl-trapping drugs to interfere with "early" events in protein modification strongly correlated with their protective potencies against allyl alcohol toxicity in hepatocytes. In mass spectrometry studies using a model lysine-containing peptide, hydralazine rapidly formed hydrazones with Michael adducts generated by acrolein. Using an antibody raised against such ternary drug-acroleinprotein complexes in Western blotting experiments, clear adduct-trapping was evident in acrolein-preloaded hepatocytes exposed to cytoprotective concentrations of hydralazine ranging from 2 to 50 M. These novel findings begin to reveal the molecular mechanisms whereby hydralazine functions as an efficient "protein adduct-trapping" drug.

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Acrolein Causes Inhibitor κB-independent Decreases in Nuclear Factor κB Activation in Human Lung Adenocarcinoma (A549) Cells

Cited by 88 publications

References 49 publications

Acrolein Inhibits Cytokine Gene Expression by Alkylating Cysteine and Arginine Residues in the NF-κB1 DNA Binding Domain

Acrolein Inhibits Cytokine Gene Expression by Alkylating Cysteine and Arginine Residues in the NF-κB1 DNA Binding Domain

Strong Protein Adduct Trapping Accompanies Abolition of Acrolein-Mediated Hepatotoxicity by Hydralazine in Mice

Protein Adduct-Trapping by Hydrazinophthalazine Drugs: Mechanisms of Cytoprotection Against Acrolein-Mediated Toxicity

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