大鼠关节软骨细胞中渗透压敏感的 TREK 通道:表达和功能作用。

Osmotically Sensitive TREK Channels in Rat Articular Chondrocytes: Expression and Functional Role.

机构信息

Department of Physiology, Biophysics and Neurosciences, Center for Research and Advanced Studies of the National Polytechnic Institute (CINVESTAV-IPN), Ciudad de México 07360, Mexico.

出版信息

Int J Mol Sci. 2024 Jul 18;25(14):7848. doi: 10.3390/ijms25147848.

Abstract

Articular chondrocytes are the primary cells responsible for maintaining the integrity and functionality of articular cartilage, which is essential for smooth joint movement. A key aspect of their role involves mechanosensitive ion channels, which allow chondrocytes to detect and respond to mechanical forces encountered during joint activity; nonetheless, the variety of mechanosensitive ion channels involved in this process has not been fully resolved so far. Because some members of the two-pore domain potassium (K2P) channel family have been described as mechanosensors in other cell types, in this study, we investigate whether articular chondrocytes express such channels. RT-PCR analysis reveals the presence of TREK-1 and TREK-2 channels in these cells. Subsequent protein expression assessments, including Western blotting and immunohistochemistry, confirm the presence of TREK-1 in articular cartilage samples. Furthermore, whole-cell patch clamp assays demonstrate that freshly isolated chondrocytes exhibit currents attributable to TREK-1 channels, as evidenced by activation by arachidonic acid (AA) and ml335 and further inhibition by spadin. Additionally, exposure to hypo-osmolar shock activates currents, which can be attributed to the presence of TREK-1 channels, as indicated by their inhibition with spadin. Therefore, these findings highlight the expression of TREK channels in rat articular chondrocytes and suggest their potential involvement in regulating the integrity of cartilage extracellular matrix.

摘要

关节软骨细胞是维持关节软骨完整性和功能的主要细胞,这对于关节的顺畅运动至关重要。它们的作用之一涉及机械敏感离子通道,这些通道使软骨细胞能够检测和响应关节活动中遇到的机械力;然而,到目前为止,参与这一过程的机械敏感离子通道的多样性尚未完全确定。由于双孔域钾(K2P)通道家族的某些成员已在其他细胞类型中被描述为机械感受器,因此在这项研究中,我们研究了关节软骨细胞是否表达这些通道。RT-PCR 分析显示这些细胞中存在 TREK-1 和 TREK-2 通道。随后的蛋白表达评估,包括 Western blot 和免疫组织化学,证实了关节软骨样本中 TREK-1 的存在。此外,全细胞膜片钳实验表明,分离的软骨细胞存在归因于 TREK-1 通道的电流,这可以通过 AA 和 ml335 的激活以及 spadin 的进一步抑制来证明。此外,低渗冲击暴露会激活电流,这归因于 TREK-1 通道的存在,如 spadin 的抑制所表明的那样。因此,这些发现强调了 TREK 通道在大鼠关节软骨细胞中的表达,并表明它们可能参与调节软骨细胞外基质的完整性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b741/11277475/a41eb3b5dedb/ijms-25-07848-g001.jpg

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