Henao Juan C, Grismaldo Adriana, Barreto Alfonso, Rodríguez-Pardo Viviana M, Mejía-Cruz Claudia Camila, Leal-Garcia Efrain, Pérez-Núñez Rafael, Rojas Patricio, Latorre Ramón, Carvacho Ingrid, Torres Yolima P
Departamento de Nutrición y Bioquímica, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, Colombia.
Grupo de Inmunobiología y Biología Celular, Departamento de Microbiología, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, Colombia.
Front Cell Dev Biol. 2021 Feb 4;9:592946. doi: 10.3389/fcell.2021.592946. eCollection 2021.
Various families of ion channels have been characterized in mesenchymal stem cells (MSCs), including some members of transient receptor potential (TRP) channels family. TRP channels are involved in critical cellular processes as differentiation and cell proliferation. Here, we analyzed the expression of TRPM8 channel in human bone marrow MSCs (hBM-MSCs), and its relation with osteogenic differentiation. Patch-clamp recordings showed that hBM-MSCs expressed outwardly rectifying currents which were increased by exposure to 500 μM menthol and were partially inhibited by 10 μM of BCTC, a TRPM8 channels antagonist. Additionally, we have found the expression of TRPM8 by RT-PCR and western blot. We also explored the TRPM8 localization in hBM-MSCs by immunofluorescence using confocal microscopy. Remarkably, hBM-MSCs treatment with 100 μM of menthol or 10 μM of icilin, TRPM8 agonists, increases osteogenic differentiation. Conversely, 20 μM of BCTC, induced a decrease of osteogenic differentiation. These results suggest that TRPM8 channels are functionally active in hBM-MSCs and have a role in cell differentiation.
间充质干细胞(MSCs)中已鉴定出多种离子通道家族,包括瞬时受体电位(TRP)通道家族的一些成员。TRP通道参与细胞分化和增殖等关键细胞过程。在此,我们分析了瞬时受体电位香草酸亚型8(TRPM8)通道在人骨髓间充质干细胞(hBM-MSCs)中的表达及其与成骨分化的关系。膜片钳记录显示,hBM-MSCs表达外向整流电流,暴露于500μM薄荷醇时该电流增加,而10μM的TRPM8通道拮抗剂BCTC可部分抑制该电流。此外,我们通过逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测到了TRPM8的表达。我们还使用共聚焦显微镜通过免疫荧光法探究了hBM-MSCs中TRPM8的定位。值得注意的是,用100μM薄荷醇或10μM异柠檬素(TRPM8激动剂)处理hBM-MSCs可增加成骨分化。相反,20μM的BCTC会导致成骨分化减少。这些结果表明,TRPM8通道在hBM-MSCs中具有功能活性,并在细胞分化中发挥作用。