Sayehmiri Fatemeh, Ilkhanizadeh-Qomi Mohsen, Naderi Nima, Monteil Arnaud, Sayyah Mohammad, Hasanzadeh Leila, Golkar Majid, Pourbadie Hamid Gholami
Skull Base Research Center, Loghman Hakim Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Neuroscience Research Center, Institute of Neuroscience and Cognition, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
J Mol Neurosci. 2025 Aug 14;75(3):107. doi: 10.1007/s12031-025-02396-7.
Kv2.1 channels, a subset of voltage-gated potassium channels, play critical roles in regulating cellular processes such as proliferation and apoptosis. While cannabidiol (CBD), a non-psychoactive phytocannabinoid, is known to modulate various ion channels, its specific effects on Kv2.1 channels remain largely unexplored. In this study, we investigated the influence of CBD on Kv2.1 channel activity and its impact on cell viability under both normal and stress conditions. To achieve stable Kv2.1 expression, HEK293 cells were transfected using the Sleeping Beauty transposon XB100 system. Puromycin (4 µg/mL) was used for selection over multiple passages. Cell viability and morphological changes were assessed using MTT assays and Giemsa staining under standard culture conditions (DMEM) and nutrient deprivation (ND) to simulate metabolic stress. CBD was applied in concentrations ranging from 3 to 3000 nM. Under standard conditions, CBD did not significantly affect cell viability during early exposure. However, under ND conditions, CBD-treated cells exhibited marked morphological deterioration and decreased viability, with these effects becoming more pronounced at higher CBD concentrations. Interestingly, Kv2.1-expressing cells showed improved baseline viability under ND, suggesting a protective role for the channel during metabolic stress. Electrophysiological analyses revealed that CBD inhibits Kv2.1 channel activity, primarily through enhanced channel inactivation. This inhibition increased cellular vulnerability to stress-induced damage. These findings reveal a dose-dependent interaction between CBD and Kv2.1 suggesting that Kv2.1 may be a relevant therapeutic target in pathological conditions such as tumor microenvironments, where cells experience oxidative stress and nutrient deprivation.
Kv2.1通道是电压门控钾通道的一个子集，在调节细胞增殖和凋亡等细胞过程中发挥着关键作用。虽然大麻二酚（CBD）是一种无精神活性的植物大麻素，已知其可调节多种离子通道，但其对Kv2.1通道的具体作用在很大程度上仍未得到探索。在本研究中，我们研究了CBD对Kv2.1通道活性的影响及其在正常和应激条件下对细胞活力的影响。为实现Kv2.1的稳定表达，使用睡美人转座子XB100系统转染HEK293细胞。在多次传代过程中使用嘌呤霉素（4μg/mL）进行筛选。在标准培养条件（DMEM）和营养剥夺（ND）下，使用MTT试验和吉姆萨染色评估细胞活力和形态变化，以模拟代谢应激。CBD的应用浓度范围为3至3000 nM。在标准条件下，早期暴露时CBD对细胞活力没有显著影响。然而，在ND条件下，CBD处理的细胞表现出明显的形态恶化和活力下降，且这些影响在较高的CBD浓度下更为明显。有趣的是，表达Kv2.1的细胞在ND下显示出改善的基线活力，表明该通道在代谢应激期间具有保护作用。电生理分析表明，CBD抑制Kv2.1通道活性，主要是通过增强通道失活。这种抑制增加了细胞对应激诱导损伤的易感性。这些发现揭示了CBD与Kv2.1之间的剂量依赖性相互作用，表明在肿瘤微环境等病理条件下，Kv2.1可能是一个相关的治疗靶点，在这些条件下细胞会经历氧化应激和营养剥夺。
