NMR Spectroscopy Identifies Chemicals in Cigarette Smoke Condensate That Impair Skeletal Muscle Mitochondrial Function

Abstract

Tobacco smoke-related diseases such as chronic obstructive pulmonary disease (COPD) are associated with high healthcare burden and mortality rates. Many COPD patients were reported to have muscle atrophy and weakness, with several studies suggesting intrinsic muscle mitochondrial impairment as a possible driver of this phenotype. Whereas much information has been learned about muscle pathology once a patient has COPD, little is known about how active tobacco smoking might impact skeletal muscle physiology or mitochondrial health. In this study, we examined the acute effects of cigarette smoke condensate (CSC) on muscle mitochondrial function and hypothesized that toxic chemicals present in CSC would impair mitochondrial respiratory function. Consistent with this hypothesis, we found that acute exposure of muscle mitochondria to CSC caused a dose-dependent decrease in skeletal muscle mitochondrial respiratory capacity. Next, we applied an analytical nuclear magnetic resonance (NMR)-based approach to identify 49 water-soluble and 12 lipid-soluble chemicals with high abundance in CSC. By using a chemical screening approach in the Seahorse XF96 analyzer, several CSC-chemicals, including nicotine, o-Cresol, phenylacetate, and decanoic acid, were found to impair ADP-stimulated respiration in murine muscle mitochondrial isolates significantly. Further to this, several chemicals, including nicotine, o-Cresol, quinoline, propylene glycol, myo-inositol, nitrosodimethylamine, niacinamide, decanoic acid, acrylonitrile, 2-naphthylamine, and arsenic acid, were found to significantly decrease the acceptor control ratio, an index of mitochondrial coupling efficiency.

Skeletal Muscle Mitochondrial Dysfunction in Chronic Obstructive Pulmonary Disease: Underlying Mechanisms and Physical Therapy Perspectives

“…Previous studies indicate that impaired mitochondrial respiration predominantly manifests as reduced mitochondrial complex I respiration efficiency [23]. Recent experiments in mice exposed to tobacco smoke also confirm that smoke exposure significantly impairs complex I-mediated MRF in a dose-dependent manner [89]. These results correspond to the findings of Brønstad et al The improvement of skeletal-muscle MRF in patients with COPD by exercise is predominantly due to increased mitochondrial mass and improved mitochondrial complex I activity [71].…”

Nontargeted Analysis in Tobacco Research: Challenges and Opportunities

“…Cigarette smoke is a highly complex mixture of chemical compounds that contains over 7000 compounds among which there are known toxicants to human health, while many more compounds are still unidentified. , Despite this complexity, various research groups have attempted to comprehensively analyze cigarette smoke composition using NTA. Some studies focused on the particulate phase, − , while others focused on the volatile components of the smoke. ,,, To reduce the complexity of the sample matrix, researchers used sample pretreatment that separates the tobacco smoke sample into different fractions such as acidic, basic, and neutral fractions. − …”

“…Therefore, to expand the range of detectable analytes, particularly polar compounds that may not be efficiently ionized by LDI, ESI was incorporated into the analytical approach . Also, nuclear magnetic resonance (NMR) spectroscopy has been utilized to profile the chemical composition of cigarette smoke condensate . The NMR method led to the detection of 49 water-soluble and 12 lipid-soluble chemicals that were highly abundant in cigarette smoke.…”

“…Furthermore, NMR sampling boasts high reproducibility due to its advanced automation, which sets it apart from MS methods. , Nevertheless, it has limitations when it comes to identifying highly volatile gaseous state compounds and those lacking protons. Moreover, NMR exhibits lower sensitivity compared to MS, making it less effective in identifying compounds at concentrations in the μM or nM range. , …”

“…al (2022) used a combination of tools and resources such as Chenomx NMR Suite software, Biological Magnetic Resonance Data Bank, Human Metabolome Database, a set of 2D experiments, and published reports to identify compounds in cigarette smoke. 33 Semiquantification. The use of specific standards and internal standards for the quantification of individual compounds or families of compounds during NTA in tobacco testing is challenging and resource-intensive.…”