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Abstract：
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 TRPV1, TRPV3, TRPV4 and TRPM8 in the MG at the gene and the protein level. RT-PCR also showed gene expression of TRPV2 but not TRPA1. 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.
摘要：
虽然干眼症（DED）中热敏瞬时受体电位通道（TRP）的参与已经知道多年，它们在睑板腺（MG）中的表达从未被研究过。本研究旨在显示它们在MG脂肪生成中的表达和参与,为DED的治疗提供可能的新的药物靶点。我们的RT-PCR,Westernblot和免疫荧光分析表明TRPV1，TRPV3，TRPV4和TRPM8在MG中的表达在基因和蛋白水平。RT-PCR还显示TRPV2而不是TRPA1的基因表达。在永生化人睑板腺上皮细胞系（hMGEC）上进行的钙成像和平面膜片钳显示，在应用辣椒素（TRPV1）或icilin（TRPM8）后，全细胞电流增加。在应用AMG9810(TRPV1)或AMTB(TRPM8)之后,可以记录降低的全细胞电流。hMGECs上的油红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。我们得出的结论是，热TRPs在MGs中的基因和蛋白质水平表达。此外，可以证明TRPV1和TRPM8的功能表达及其对脂质表达的贡献。因此,TRP是潜在的药物靶标，其在睑板腺功能障碍治疗中的临床意义需要进一步研究。
