Menthol causes mitochondrial Ca2+-influx, affects structure-function relationship and cools mitochondria

Menthol is a small bioactive compound able to cause several physiological changes and has multiple molecular targets. Therefore, cellular response against menthol is complex, and still poorly understood. In this work, we used a human osteosarcoma cell line (Saos-2) and analysed the effect of menthol, especially in terms of cellular, subcellular and molecular aspects. We demonstrate that menthol causes increased mitochondrial Ca2+ in a complex manner, which is mainly contributed by intracellular sources, including ER. Menthol also changes the Ca2+-load of individual mitochondrial particles in different conditions. Menthol increases ER-mito contact points, causes mitochondrial morphological changes, and increases mitochondrial ATP, cardiolipin, mitochondrial ROS and reduces mitochondrial membrane potential (Delta psi m). Menthol also prevents the mitochondrial quality damaged by sub-lethal and lethal doses of CCCP. In addition, menthol lowers the mitochondrial temperature within cell and also serves as a cooling agent for the isolated mitochondria in a cell free system too. Notably, menthol-induced reduction of mitochondrial temperature is observed in diverse types of cells, including neuronal, immune and cancer cells. As the higher mitochondrial temperature is a hallmark of several inflammatory, metabolic, disease and age-related disorders, we propose that menthol can serve as an active anti-aging compound against all these disorders. These findings may have relevance in case of several pharmacological and clinical applications of menthol.Significance statement: Menthol is a plant-derived bioactive compound that is widely used for several physio-logical, behavioural, addictive, and medicinal purposes. It is a well-established cooling and analgesic agent. However, the exact cellular and sub-cellular responses of menthol is poorly understood. In this work, we have characterized the effects of menthol on mitochondrial metabolism. Menthol regulates mitochondrial Ca2+, ATP, superoxides, cardiolipin, membrane-potential, and ER-mito contact sites. Moreover, the cooling agent menthol also cools down mitochondria and protects mitochondrial damage by certain toxins. These findings may promote use of menthol as a useful supplementary agent for anti-aging, anti-cancer, anti-inflammatory purposes where higher mitochondrial temperature is prevalent.

Automated summary

Menthol is a bioactive compound that causes various physiological changes, and its cellular and subcellular effects are still poorly understood. In this study, the effects of menthol on mitochondrial metabolism were characterized. Menthol regulates mitochondrial Ca2+, ATP, superoxides, cardiolipin, membrane potential, and ER-mito contact sites. Additionally, menthol acts as a cooling agent for mitochondria and protects against mitochondrial damage from certain toxins. These findings suggest that menthol could be used as a supplementary agent for anti-aging, anti-cancer, and anti-inflammatory purposes where higher mitochondrial temperature is prevalent.