Mass Spectrometric Approaches to the Identification of Potential Ingredients in Cigarette Smoke Causing Cytotoxicity

Abstract

Cigarette smoke contains many harmful chemicals that contribute to the pathogenesis of smoking-related diseases such as chronic obstructive pulmonary disease, cancer, and cardiovascular disease. Many studies have been done to identify cytotoxic chemicals in cigarette smoke and elucidate the onset of the above-mentioned diseases caused by smoking. However, definitive mechanisms for cigarette smoke toxicity remain unknown. As candidates for cytotoxic chemicals, we have recently found methyl vinyl ketone (MVK) and acetic anhydride in nicotine/tar-free cigarette smoke extract (CSE) using L-tyrosine (Tyr), an amino acid with highly reactive hydroxyl group. The presence of MVK and acetic anhydride in CSE was confirmed by gas chromatography-mass spectrometry (GC/MS). We also found new reaction products formed in B16-BL6 mouse melanoma (B16-BL6) cells treated with CSE using LC/MS. These were identified as glutathione (GSH) conjugates of α,β-unsaturated carbonyl compounds, MVK, crotonaldehyde (CA), and acrolein (ACR), by the mass value and product ion spectra of these new products. ACR and MVK are type-2 alkenes, which are well known as electron acceptors and form Michael-type adducts to nucleophilic side chain of amino acids on peptides. These α,β-unsaturated carbonyl compounds may have a key role in CSE-induced cell death.

Relationships among smoking, oxidative stress, inflammation, macromolecular damage, and cancer

“…Hydroxypropanodeoxyguanosine (HOPdG) is the predominant promutagenic acrolein-DNA adduct [79,107,108]. Binding of acrolein to intracellular GSH results in depletion of the supply of this antioxidant [109]. 4-HNE is another highly reactive aldehyde, capable of binding to GSH via GST activity, resulting in decreased antioxidant capacity [110,111].…”

Assessment of AcuteIn VitroHuman Cellular Responses to Smoke Extracts from a Reduced Toxicant Prototype Cigarette

“…Carbonyls such as acrolein have been linked to the induction of cellular oxidative stress 24 and it has been reported that carbonyls are captured in AqE. 23,[25][26][27][28] Furthermore, the ratio of carbonyl components of AqE are similar to that observed in cigarette smoke, although the absolute levels were lower. 29 The antioxidant status of NCI-H292 lung epithelial cells was assessed using the GSH-GloÔ assay to measure changes in the level of the reduced form of the antioxidant glutathione.…”

“…Literature shows that the vapor phase is primarily responsible for the cytotoxic effects observed in this assay. 25,[30][31][32] The RTP cigarette had significant reductions in acetaldehyde and acrolein ( Supplementary Table S3) compared with the CC cigarette, both of which have been shown to inhibit cell proliferation and/or induce cytotoxicity in vitro. 12,[33][34][35][36][37] This may explain why WS from RTP cigarettes induced significantly less cytotoxicity in lung epithelial cells versus the CC cigarettes.…”

Oxidative, inflammatory, genetic, and epigenetic biomarkers associated with chronic obstructive pulmonary disorder

“…To remove NO and unstable superoxide anion both (cause inflammation) rapidly, they are scavenged by oxyhaemoglobin present in red blood cells. In phagocytes common ROS is NO, and when NO combines with superoxide anion, it produce peroxynitrite, which can target protein thiols and tyrosine (Miller et al, 2013), whereas unstable superoxide anion can produce superoxide dismutase (SOD) producing hydrogen peroxide (Horiyama, Kunitomo, Yoshikawa, & Nakamura, 2016).…”

“…To remove NO and unstable superoxide anion both (cause inflammation) rapidly, they are scavenged by oxyhaemoglobin present in red blood cells. In phagocytes common ROS is NO, and when NO combines with superoxide anion, it produce peroxynitrite, which can target protein thiols and tyrosine (Miller et al, ), whereas unstable superoxide anion can produce superoxide dismutase (SOD) producing hydrogen peroxide (Horiyama, Kunitomo, Yoshikawa, & Nakamura, ). Hydrogen peroxide participates in various reactions to produce other oxidant species, for example, reactive nitrogen species and reactive carbon species by Fenton reaction or by the production of HOCl by utilizing myeloperoxidase (Dekhuijzen et al, ; Marotta et al, ).…”