Vascular Endothelial Growth Factor (VEGF) Induced Downstream Responses to Transient Receptor Potential Vanilloid 1 (TRPV1) and 3-Iodothyronamine (3-T1AM) in Human Corneal Keratocytes
Vascular Endothelial Growth Factor (VEGF) Induced Downstream Responses to Transient Receptor Potential Vanilloid 1 (TRPV1) and 3-Iodothyronamine (3-T1AM) in Human Corneal Keratocytes
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DOI：
10.3389/fendo.2018.00670
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发表时间：
2018-11
影响因子：
5.2
通讯作者：
Ersal Türker;F. Garreis;N. Khajavi;P. Reinach;P. Joshi;T. Brockmann;Alexander Lucius;Nina Ljubojevic;Elizabeth Turan;D. Cooper;F. Schick;Rob Reinholz;U. Pleyer;J. Köhrle;S. Mergler
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文献类型：
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作者：
Ersal Türker;F. Garreis;N. Khajavi;P. Reinach;P. Joshi;T. Brockmann;Alexander Lucius;Nina Ljubojevic;Elizabeth Turan;D. Cooper;F. Schick;Rob Reinholz;U. Pleyer;J. Köhrle;S. Mergler
本研究旨在确定瞬时受体电位(Trp)、褪黑素8(TRPM8)、色氨酸香草素1(TRPV1)和血管内皮生长因子(VEGF)受体三联体之间的串扰是否对血管内皮生长因子(VEGF)诱导的人角膜基质细胞钙信号传导有调节作用。应用逆转录聚合酶链式反应、定量聚合酶链式反应和免疫组织化学方法，检测了人角膜基质细胞系(HCK)和人角膜切片中TRPV1和TRPM8基因和蛋白的共表达。使用光电倍增管和单细胞数字成像系统的荧光钙成像以及平面膜片钳技术测量了相对细胞内钙水平和潜在的全细胞电流。TRPV1激动剂辣椒素增加细胞内钙离子水平和全细胞电流，而拮抗剂卡萨西平(CPZ)则抑制这两种作用。CPZ可抑制血管内皮生长因子诱导的钙瞬变和全细胞电流的升高，而选择性TRPM8拮抗剂AMTB可增加血管内皮生长因子的信号转导。相反，内源性甲状腺激素衍生代谢物3-碘甲状腺原胺(3-T1AM)抑制了血管内皮生长因子诱导的电流的增加。TRPM8激动剂薄荷醇增加电流，而AMTB抑制这一反应。3-T1AM诱导的TRPM8激活可抑制VEGFR和TRPV1之间的串扰，而VEGFR和TRPV1之间的串扰可抑制VEGFR和TRPV1之间的串扰，从而导致血管内皮生长因子诱导的钙内流及其离子电流的增加。这种抑制反过来又阻断了血管内皮生长因子诱导的TRPV1的激活。因此，TRPM8和TRPV1之间的串扰抑制了VEGFR诱导的TRPV1的激活。
This study was undertaken to determine if crosstalk among the transient receptor potential (TRP) melastatin 8 (TRPM8), TRP vanilloid 1 (TRPV1), and vascular endothelial growth factor (VEGF) receptor triad modulates VEGF-induced Ca2+ signaling in human corneal keratocytes. Using RT-PCR, qPCR and immunohistochemistry, we determined TRPV1 and TRPM8 gene and protein coexpression in a human corneal keratocyte cell line (HCK) and human corneal cross sections. Fluorescence Ca2+ imaging using both a photomultiplier and a single cell digital imaging system as well as planar patch-clamping measured relative intracellular Ca2+ levels and underlying whole-cell currents. The TRPV1 agonist capsaicin increased both intracellular Ca2+ levels and whole-cell currents, while the antagonist capsazepine (CPZ) inhibited them. VEGF-induced Ca2+ transients and rises in whole-cell currents were suppressed by CPZ, whereas a selective TRPM8 antagonist, AMTB, increased VEGF signaling. In contrast, an endogenous thyroid hormone-derived metabolite 3-Iodothyronamine (3-T1AM) suppressed increases in the VEGF-induced current. The TRPM8 agonist menthol increased the currents, while AMTB suppressed this response. The VEGF-induced increases in Ca2+ influx and their underlying ionic currents stem from crosstalk between VEGFR and TRPV1, which can be impeded by 3-T1AM-induced TRPM8 activation. Such suppression in turn blocks VEGF-induced TRPV1 activation. Therefore, crosstalk between TRPM8 and TRPV1 inhibits VEGFR-induced activation of TRPV1.