Kook Hyun, Itoh Hiroshi, Choi Bong Seok, Sawada Naoki, Doi Kentaro, Hwang Tae Ju, Kim Kyung Keun, Arai Hiroshi, Baik Yung Hong, Nakao Kazuwa
Research Institute of Medical Sciences, Chonnam National University Medical School, Gwangju 501-746, Republic of Korea.
Am J Physiol Heart Circ Physiol. 2003 Apr;284(4):H1388-97. doi: 10.1152/ajpheart.00414.2002. Epub 2002 Dec 27.
Both nitric oxide (NO) and natriuretic peptides produce apoptosis of vascular smooth muscle cells. However, there is evidence that NO induces endothelial cell proliferation, which suggests that there is a difference in the response of endothelial cells to natriuretic peptides. The purpose of this study was to investigate the effect of atrial natriuretic peptide (ANP) on human endothelial cell survival. ANP within the physiological concentration (10(-11) mol/l) induced a 52% increase in the number of human coronary arterial endothelial cells and a 63% increase in human umbilical vein endothelial cells at a low concentration of serum. The increase in cell numbers was blocked by pretreatment with RP8-CPT-cGMP (RP8), a cGMP-dependent protein kinase inhibitor, with wortmannin, an Akt/PKB inhibitor, and with PD-98059, an ERK1/2 inhibitor. In a Transwell migration test, ANP also increased the cell migration, and RP8, wortmannin, and PD-98059 blocked this increase. A wound healing assay was performed to examine the effects of ANP on regeneration in vitro. ANP increased both cell numbers and migration, but the effects were blocked by the above three kinase inhibitors. ANP increased the expression of phospho-Akt and of phospho-ERK1/2 within 1.5 h. These results suggest that ANP can potentiate endothelial regeneration by cGMP-dependent protein kinase stimulation and subsequent Akt and ERK1/2 activations.
一氧化氮（NO）和利钠肽均可诱导血管平滑肌细胞凋亡。然而，有证据表明NO可诱导内皮细胞增殖，这提示内皮细胞对利钠肽的反应存在差异。本研究的目的是探讨心房利钠肽（ANP）对人内皮细胞存活的影响。在低血清浓度下，生理浓度（10⁻¹¹ mol/l）的ANP可使人类冠状动脉内皮细胞数量增加52%，使人类脐静脉内皮细胞数量增加63%。细胞数量的增加被cGMP依赖性蛋白激酶抑制剂RP8-CPT-cGMP（RP8）、Akt/PKB抑制剂渥曼青霉素和ERK1/2抑制剂PD-98059预处理所阻断。在Transwell迁移试验中，ANP也增加了细胞迁移，且RP8、渥曼青霉素和PD-98059可阻断这种增加。进行伤口愈合试验以检测ANP对体外再生的影响。ANP增加了细胞数量和迁移，但上述三种激酶抑制剂可阻断这些作用。ANP在1.5小时内增加了磷酸化Akt和磷酸化ERK1/2的表达。这些结果表明，ANP可通过刺激cGMP依赖性蛋白激酶以及随后激活Akt和ERK1/2来增强内皮细胞再生。
