Small cytoplasmic domain peptides of natriuretic peptide receptor-C attenuate cell proliferation through Giα protein/MAP kinase/PI3-kinase/AKT pathways
Small cytoplasmic domain peptides of natriuretic peptide receptor-C attenuate cell proliferation through Giα protein/MAP kinase/PI3-kinase/AKT pathways
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DOI：
10.1152/ajpheart.00327.2006
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发表时间：
2006-12-01
影响因子：
4.8
通讯作者：
Anand-Srivastava, Madhu B.
中科院分区：
文献类型：
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作者：
Hashim, Shehla;Li, Yuan;Anand-Srivastava, Madhu B.
本研究观察了C-心房钠尿肽（ANP）4-23和来自钠尿肽受体（NPR）-C胞浆区不同区域的几个含12个氨基酸的肽段在血管紧张素Ⅱ（ANG）Ⅱ、内皮素（ET）-1、和精氨酸加压素（AVP）。所用的肽片段具有完整的G（i）激活剂序列（KH 472）-H-461（肽1）和H-481-H-492（肽3）或分别具有截短的COOH或NH 2末端的部分Gi激活剂序列R469-K480（肽2）和I-465-H-472（肽Y）。使用的另一种肽没有结构特异性（Q（473）-K-480，肽X），或者是肽1（肽Z）的乱序肽对照。通过[H-3]胸苷掺入测定，ANG II、ET-1和AVP显著刺激这些细胞中的DNA合成，肽1、2和3以浓度依赖性方式抑制，而肽X、Y和Z则不抑制，表观Ki在1和10 nM之间。此外，与NPR-C相互作用的C-ANP（4-23）也抑制由血管活性肽刺激的DNA合成;然而，C-ANP（4-23）引起的抑制与肽1引起的抑制不相加。另一方面，这些细胞中的基础DNA合成不受C-ANP（4-23）或肽片段的抑制。此外，PD-98059和渥曼青霉素可抑制血管活性肽诱导的DNA合成刺激，肽1可增强这种抑制作用。此外，肽1还抑制血管活性肽诱导的ERK 1/2和AKT磷酸化，并增强G（i）α蛋白的表达。这些数据表明，C-ANP（4-23）和含有12个氨基酸的小肽片段（与NPR-C的胞质结构域区域无关）通过G（i）α蛋白和MAP激酶/磷脂酰肌醇3-激酶/AKT途径抑制血管活性肽的增殖反应。
The present studies were undertaken to investigate the effect of C-atrial natriuretic peptide (ANP) 4-23 and several peptide fragments containing 12 amino acids from different regions of the cytoplasmic domain of natriuretic peptide receptor (NPR)-C on cell proliferation in the absence or presence of angiotensin (ANG) II, endothelin ( ET)-1, and arginine vasopressin (AVP) in A-10 vascular smooth muscle cells (VSMC). The peptide fragments used have either complete G(i) activator sequences (KH472)-H-461 ( peptide 1) and H-481-H-492 ( peptide 3) or partial Gi activator sequences R469-K480 ( peptide 2) and I-465-H-472 ( peptide Y) with truncated COOH or NH2 terminus, respectively. The other peptide used had no structural specificity (Q(473)-K-480, peptide X) or was the scrambled peptide control for peptide 1 ( peptide Z). ANG II, ET-1 and AVP significantly stimulated DNA synthesis in these cells as determined by [H-3] thymidine incorporation that was inhibited by peptides 1, 2, and 3 and not by peptides X, Y, and Z in a concentration-dependent manner, with an apparent K-i between 1 and 10 nM. In addition, C-ANP(4-23), which interacts with NPR-C, also inhibited DNA synthesis stimulated by vasoactive peptides; however, the inhibition elicited by C-ANP(4-23) was not additive with the inhibition elicited by peptide 1. On the other hand, basal DNA synthesis in these cells was not inhibited by C-ANP(4-23) or the peptide fragments. Furthermore, vasoactive peptide-induced stimulation of DNA synthesis was inhibited by PD-98059 and wortmannin, and this inhibition was potentiated by peptide 1. In addition, peptide 1 also inhibited vasoactive peptide-induced phosphorylation of ERK1/2 and AKT and enhanced expression of G(i)alpha proteins. These data suggest that C-ANP(4-23) and small peptide fragments containing 12 amino acids irrespective of the region of the cytoplasmic domain of NPR-C inhibit proliferative responses of vasoactive peptides through G(i)alpha protein and MAP kinase/phosphatidylinositol 3-kinase/AKT pathways.