Department of Functional and Clinical Anatomy, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Universitätsstraße 19, 91054 Erlangen, Germany.
Department of Ophthalmology, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 13353 Berlin, Germany.
Int J Mol Sci. 2024 Apr 5;25(7):4043. doi: 10.3390/ijms25074043.
While the involvement of thermosensitive transient receptor potential channels (TRPs) in dry eye disease (DED) has been known for years, their expression in the meibomian gland (MG) has never been investigated. This study aims to show their expression and involvement in the lipogenesis of the MG, providing a possible new drug target in the treatment of DED. Our RT-PCR, Western blot and immunofluorescence analysis showed the expression of , , and in the MG at the gene and the protein level. RT-PCR also showed gene expression of but not . Calcium imaging and planar patch-clamping performed on an immortalized human meibomian gland epithelial cell line (hMGECs) demonstrated increasing whole-cell currents after the application of capsaicin (TRPV1) or icilin (TRPM8). Decreasing whole-cell currents could be registered after the application of AMG9810 (TRPV1) or AMTB (TRPM8). Oil red O staining on hMGECs showed an increase in lipid expression after TRPV1 activation and a decrease after TRPM8 activation. We conclude that thermo-TRPs are expressed at the gene and the protein level in MGs. Moreover, TRPV1 and TRPM8's functional expression and their contribution to their lipid expression could be demonstrated. Therefore, TRPs are potential drug targets and their clinical relevance in the therapy of meibomian gland dysfunction requires further investigation.
虽然热敏瞬时受体电位通道(TRP)参与干眼病(DED)已为人所知多年,但它们在睑板腺(MG)中的表达从未被研究过。本研究旨在展示它们在 MG 的脂生成中的表达和参与,为 DED 的治疗提供一个可能的新药物靶点。我们的 RT-PCR、Western blot 和免疫荧光分析显示, 、 、 和 在基因和蛋白质水平上在 MG 中表达。RT-PCR 还显示了基因表达 但不是 。在永生化人睑板腺上皮细胞系(hMGEC)上进行的钙成像和平板膜片钳实验表明,辣椒素(TRPV1)或异鼠李素(TRPM8)给药后会增加全细胞电流。在用 AMG9810(TRPV1)或 AMTB(TRPM8)处理后,可以记录到全细胞电流减少。在 hMGEC 上进行的油红 O 染色显示 TRPV1 激活后脂质表达增加,TRPM8 激活后脂质表达减少。我们得出结论,热敏 TRP 在 MG 中以基因和蛋白质水平表达。此外,TRPV1 和 TRPM8 的功能表达及其对脂质表达的贡献可以得到证明。因此,TRP 是潜在的药物靶点,它们在治疗睑板腺功能障碍中的临床相关性需要进一步研究。