GLP-1 inhibits VEGFA-mediated signaling in isolated human endothelial cells and VEGFA-induced dilation of rat mesenteric arteries
GLP-1 inhibits VEGFA-mediated signaling in isolated human endothelial cells and VEGFA-induced dilation of rat mesenteric arteries
复制标题
DOI：
10.1152/ajpheart.00316.2016
复制
发表时间：
2016-11-01
影响因子：
4.8
通讯作者：
Dissing, Steen
中科院分区：
文献类型：
--
作者：
Egholm, Cecilie;Khammy, Makhala Michell;Dissing, Steen
我们研究了胰升糖素样肽-1(GLP-1)、GLP-1(1-36)和GLP-1(7-36)对血管内皮生长因子-A(VEGFA)诱导的内皮依赖性信号转导和血管扩张的急性影响。我们的假设是，肠道L细胞释放的GLP-1调节与PLCγ激活、Src和内皮型一氧化氮合酶(ENOS)信号相关的过程，从而控制内皮血管张力。通过RT-PCR分析，我们在人皮肤微血管内皮细胞(HDMEC)、人视网膜微血管内皮细胞和大鼠动脉中发现了GLP-1受体(GLP-1R)的mRNA。在用血栓素类似物U46619预收缩至最大收缩的80-90%的大鼠肠系膜阻力动脉上，VEGFA(25 ng/ml)可引起小而渐进的松弛(28.9+/-3.9%)。GLP-1(1-36)(500 NM)或GLP-1(7-36)(1 NM)可阻断VEGFA引起的血管松弛。在内皮剥脱的动脉和一氧化氮合酶抑制剂N omega-硝基-L-精氨酸甲酯(100mU)预处理的动脉中，血管内皮生长因子诱导的血管松弛也被抑制。对雄性Wistar大鼠的体内研究也表明，GLP-1(7-36)可抑制VEGFA诱导的同一动脉的血管扩张。在分离的内皮细胞中，GLP-1(1-36)和GLP-1(7-36)可降低VEGFA诱导的PLC-γ的磷酸化。使用Fura-2对内皮细胞和大鼠肠系膜阻力动脉进行的钙成像显示，两种GLP-1类似物都能减少VEGFA诱导的钙信号。GLP-1(1-36)还可降低VEGFA诱导的HDMEC中eNOS的磷酸化。综上所述，GLP-1通过抑制VEGFR2介导的钙信号和内皮NO的合成来减轻VEGFA对阻力动脉的松弛作用。GLP-1本身也可诱导Src和ERK1/2的磷酸化，从而导致细胞增殖，并与血管通透性有关。
We investigated the acute effects of glucagon-like peptide-1 (GLP-1), GLP-1(1-36), and GLP-1(7-36) on vascular endothelial growth factor-A (VEGFA)-induced endothelium-dependent signaling and vasodilation. Our hypothesis was that GLP-1 released from intestinal L-cells modulates processes related to PLC gamma activation, Src, and endothelial NOS (eNOS) signaling, thereby controlling endothelial vessel tone. By using RT-PCR analysis, we found mRNA for the GLP-1 receptor (GLP-1R) in human dermal microvascular endothelial cells (HDMEC), human retinal microvascular endothelial cells, and rat arteries. In isolated rat mesenteric resistance arteries precontracted with the thromboxane analog U46619 to 80-90% of maximum contraction, VEGFA (25 ng/ml) caused a small and gradual relaxation (28.9 +/- 3.9%). Pretreatment of arteries with either GLP-1(1-36) (500 nM) or GLP-1(7-36) (1 nM) abolished the VEGFA-induced relaxation. VEGFA-induced relaxations were also inhibited in endothelial-denuded arteries and in arteries pretreated with the nitric oxide synthase (NOS) inhibitor, N omega-nitro-L-arginine methyl ester (100 mu M). In vivo studies on male Wistar rats also revealed that GLP-1(7-36) inhibited VEGFA-induced vasodilation of the same arteries. In isolated endothelial cells, GLP-1(1-36) and GLP-1(7-36) caused a reduction in VEGFA-induced phosphorylation of PLC gamma. Ca2+ imaging of endothelial cells and rat mesenteric resistance arteries using fura-2, revealed that both GLP-1 analogs caused a reduction in VEGFA-induced Ca2+ signaling. GLP-1(1-36) also reduced VEGFA-induced eNOS phosphorylation in HDMEC. In conclusion, GLP-1 reduced relaxation induced by VEGFA in resistance arteries by inhibiting VEGFR2-mediated Ca2+ signaling and endothelial NO synthesis. GLP-1, on its own, also induced phosphorylation of Src and ERK1/2 that can lead to proliferation and is implicated in vessel permeability.