薄荷醇是一种小型的生物活性化合物，能够引起多种生理变化，并具有多个分子靶点。因此，细胞对薄荷醇的反应是复杂的，并且仍然知之甚少。在本研究中，我们使用人骨肉瘤细胞系（Saos-2）分析了薄荷醇的作用，特别关注细胞、亚细胞和分子方面。我们证明薄荷醇以复杂的方式引起线粒体Ca2+增加，其中主要贡献者是细胞内来源，包括内质网。薄荷醇还会改变不同条件下单个线粒体粒子的Ca2+负载。薄荷醇增加了内质网与线粒体的接触点，引起线粒体形态学变化，并增加线粒体ATP、心磷脂、线粒体ROS，降低线粒体膜电位（ΔΨm）。薄荷醇还能阻止亚致死和致死剂量的CCCP对线粒体质量的损伤。此外，薄荷醇降低了细胞内线粒体的温度，并在无细胞体系中作为冷却剂对离体线粒体起作用。值得注意的是，薄荷醇引起的线粒体温度降低在包括神经、免疫和癌细胞在内的多种细胞中被观察到。由于更高的线粒体温度是多种炎症、代谢、疾病和与年龄相关的疾患的标志，我们提出薄荷醇可以作为一种抗衰老化合物对所有这些疾患起作用。这些发现对于薄荷醇在多种药理学和临床应用中具有重要意义。意义陈述：薄荷醇是一种天然生物活性化合物，广泛用于多种生理、行为、成瘾和医疗目的。它是一种被广泛认可的“冷却和镇痛剂”。然而，薄荷醇的确切细胞和亚细胞反应尚不清楚。在这项研究中，我们对薄荷醇对线粒体代谢的影响进行了表征。薄荷醇调节线粒体的Ca2+、ATP、超氧化物、心磷脂、膜电位和内质网-线粒体接触位点。此外，具有冷却作用的薄荷醇还能降低线粒体温度，并保护线粒体免受某些毒素的损伤。这些发现可能促进薄荷醇作为抗衰老、抗癌、抗炎症的有用辅助剂的使用，其中具有较高线粒体温度。版权所有 © 2023 Elsevier Inc.

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 (ΔΨm). 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 physiological, 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.Copyright © 2023. Published by Elsevier Inc.