Immunolocalization and expression of vascular endothelial growth factor receptors (VEGFRs) and neuropilins (NRPs) on Keratinocytes in human epidermis
Immunolocalization and expression of vascular endothelial growth factor receptors (VEGFRs) and neuropilins (NRPs) on Keratinocytes in human epidermis
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DOI：
10.2119/2006-00024.man
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发表时间：
2006-07-01
影响因子：
5.7
通讯作者：
Zheng, Min
中科院分区：
文献类型：
--
作者：
Man, Xiao-Yong;Yang, Xiao-Hrong;Zheng, Min
血管内皮生长因子（VEGF）在正常和病理性血管生成中发挥重要作用。 VEGF 受体（VEGFR，包括 VEGFR-1、VEGFR-2 和 VEGFR-3）和神经毡蛋白（NRP，包括 NRP-1 和 NRF-2）是 VEGF 的高亲和力受体，通常被认为对内皮细胞具有特异性。在这里，我们展示了培养的表皮角质形成细胞上 VEGFR 和 NRP 在 mRNA 和蛋白质水平上的表达。我们通过手术皮肤标本表皮的免疫荧光（IF）染色进一步定位这些受体。我们发现除角质层外的表皮所有层中 VEGFR 和 NRP 均呈阳性染色。 VEGFR-1和VEGFR-2主要表达于基底细胞和邻近的棘细胞角质形成细胞的细胞质膜上。除角质细胞层外，表皮所有层均表现出一致的 VEGFR-3、NRP-1 和 NRP-2 模式。 NRP-1和NRP-2的切片染色也显示出弥漫性强荧光，并且定位于角质形成细胞的细胞膜和细胞质。在另一组实验中，用不同浓度的 VEGF 处理角质形成细胞，培养物中含有或不含 VEGFR-2 中和抗体。 VEGF增强角质形成细胞的增殖和迁移，而这些作用被VEGFR-2中和抗体预处理部分抑制。 VEGF 处理可降低角质形成细胞对 IV 型胶原包被培养板的粘附，但通过 VEGFR-2 中和抗体预处理可完全逆转这种降低。综上所述，我们的结果表明，角质形成细胞上 VEGFR 和 NRP 的表达可能构成其活性的重要调节因子，并且可能负责表皮中的自分泌信号传导。
Vascular endothelial growth factor (VEGF) plays an important role in normal and pathological angiogenesis. VEGF receptors (VEGFRs, including VEGFR-1, VEGFR-2, and VEGFR-3) and neuropilins (NRPs, including NRP-1 and NRF-2) are high-affinity receptors for VEGF and are typically considered to be specific for endothelial cells. Here we showed expression of VEGFRs and NRPs on cultured epidermal keratinocytes at both mRNA and protein levels. We further localized these receptors by immunofluorescence (IF) staining in the epidermis of surgical skin specimens. We found positive staining for VEGFRs and NRPs in all layers of the epidermis except for the stratum corneum. VEGFR-1 and VEGFR-2 are primarily expressed on the cytoplasmic membrane of basal cells and the adjacent spinosum keratinocytes. All layers of the epidermis except for the horny cell layer demonstrated a uniform pattern of VEGFR-3, NRP-1, and NRP-2. Sections staining for NRP-1 and NRP-2 also showed diffuse intense fluorescence and were localized to the cell membrane and cytoplasm of keratinocytes. In another panel of experiments, keratinocytes were treated with different concentrations of VEGF, with or without VEGFR-2 neutralizing antibody in culture. VEGF enhanced the proliferation and migration of keratinocytes, and these effects were partially inhibited by pretreatment with VEGFR-2 neutralizing antibody. Adhesion of keratinocytes to type IV collagen-coated culture plates was decreased by VEGF treatment, but this reduction could be completely reversed by pretreatment with VEGFR-2 neutralizing antibody. Taken together, our results suggest that the expression of VEGFRs and NRPs on keratinocytes may constitute important regulators for its activity and may possibly be responsible for the autocrine signaling in the epidermis.