Menthol Cigarette Smoke Induces More Severe Lung Inflammation Than Non-menthol Cigarette Smoke Does in Mice With Subchronic Exposure – Role of TRPM8

Abstract

In smokers with chronic obstructive pulmonary disease, more severe lung inflammation is associated with menthol cigarette smoking compared to non-menthol cigarette smoking. However, the mechanisms remain unclear. Menthol is an activator of transient receptor potential melastatin-8 (TRPM8), which is also sensitive to reactive oxygen species (ROS). Our recent in vitro study demonstrated that the extracts of menthol cigarette smoke (M-CS) can induce greater ROS-sensitive, TRPM8-mediated, mitogen-activated protein kinase (MAPK)-dependent inflammatory responses in lung epithelial cells than the extracts of non-menthol cigarette smoke (Non-M-CS) can. In this study, we tested the hypothesis that M-CS can induce more severe lung inflammation than Non-M-CS can via the additional action of menthol in M-CS on epithelial and lung TRPM8 in mice. Compared with Non-M-CS exposure, subchronic M-CS exposure for 7 days up-regulated the epithelial and lung expression of TRPM8, induced more vigorous activation of epithelial and lung MAPKs, and caused more severe lung inflammation. The MAPK activation was evidenced by the increased expression of phosphor-extracellular signal-regulated and phosphor-c-Jun N-terminal kinases. The lung inflammation was evidenced by pathohistological findings and increases in several inflammatory indices. Moreover, treatment with a TRPM8 antagonist (N-(3-aminopropyl)-2-{[(3-methylphenyl)methyl]oxy}-N-(2-thienylmethyl)benzamide; AMTB) greatly suppressed the MAPK activation and lung inflammation induced by Non-M-CS and M-CS, and the residual responses to these two types of CS did not differ. Conversely, the levels of biomarkers of acute CS exposure (20 min), including carboxyhemoglobin and cotinine (a nicotine metabolite) in blood plasma, and superoxide and hydrogen peroxide (two major types of ROS) in bronchoalveolar lavage fluid, did not show significant differences in the mice with Non-M-CS and M-CS exposure. We concluded that M-CS could induce greater TRPM8-mediated activation of MAPKs and lung inflammation than Non-M-CS could in mice with subchronic exposure. The augmented inflammatory effects of M-CS are unlikely due to a larger total amount of CS inhaled, but may be caused by an additional stimulation of epithelial and lung TRPM8 by menthol in M-CS. A common stimulant (presumably ROS) generated by both CS types may also stimulate TRPM8, activate MAPKs, and induce lung inflammation because treatment with AMTB could reduce these responses to Non-M-CS.

Reversal of NADPH Oxidase-Dependent Early Oxidative and Inflammatory Responses in Chronic Obstructive Pulmonary Disease by Puerarin

“…In our study, superoxide production was found significantly increased in mouse lung tissues harvested 2 hours after the 1 hour ACS exposure, and most dramatically increased at 6 hours post exposure. Our data indicate a time-dependent production of superoxide in ACS exposed mouse lung tissues, which is consistent with previous observations evidencing ACS induction of superoxide and hydrogen peroxide accumulation in BAL fluid [ 35 ]. Of note, here, we elucidated that Puerarin dose-dependently abolished ACS induced superoxide production in the mouse lung tissues.…”

E-cigarette vape and lung ACE2 expression: Implications for coronavirus vulnerability

“…Further, a large proportion of youth preferentially consume flavored nicotine products, with menthol as one of the most popular for tobacco cigarettes ( Jackson et al, 2020 ; Villanti et al, 2017 ). In smokers with chronic obstructive pulmonary disease, more severe lung inflammation is associated with menthol cigarette smoking compared to non-mentholated products ( Park et al, 2015 ), and in mice, sub-chronic exposure to the extracts of menthol cigarette smoke induces lung inflammation ( Lin et al, 2018 ). Although the putative action of menthol and other flavorants on ACE2 has not yet been elucidated, one may predict that menthol would act synergistically with nicotine to exert detrimental effects, although this needs to be investigated in further studies.…”

Menthol in electronic cigarettes: A contributor to respiratory disease?

“…Pure menthol was cytotoxic to bronchial epithelium at the concentrations found in EC products when tested in vitro with the MTT assay using 2D submerged cell cultures (Behar et al, 2017;Hua et al, 2019). Lin et al, (2018) showed that subchronic exposure of mice to mentholated cigarette smoke induced more inflammation in lungs than smoke from nonmentholated cigarettes. Recently, serious respiratory illness and death have been attributed to EC use, and patients requiring hospitalization have been reported to have "e-cigarette or vaping product use-associated lung injury" (EVALI) (Centers for Disease Control and Prevention, 2019).…”