精氨酸升压素(arginine vasopressin, AVP)是高血压和心力衰竭时激活的神经体液和血流动力学因子,同时,它还具有直接的生长刺激作用。我们以往的研究显示AVP可诱导新生大鼠心肌成纤维细胞(cardiac fibroblasts, CFs)增殖。本研究旨在进一步观察AVP是否对成年大鼠CFs增殖具有促
Acta Physiologica Sinica, June 25, 2008, 60 (3): 333-340
333
Research Paper
Arginine vasopressin stimulates proliferation of adult rat cardiac fibroblastsvia protein kinase C-extracellular signal-regulated kinase 1/2 pathway
HE Yan-Ping, ZHAO Lian-You*, ZHENG Qiang-Sun, LIU Shao-Wei, ZHAO Xiao-Yan, LU Xiao-Long,NIU Xiao-Lin
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Department of Cardiology, Tangdu Hospital, the Fourth Military Medical University, Xi’an 710038, China
Abstract: Arginine vasopressin (AVP), a neurohormone and hemodynamic factor implicated in the pathophysiology of hypertensionand congestive heart failure, can also act as a growth-stimulating factor. Our previous work demonstrated that AVP is a mitogen forneonatal rat cardiac fibroblasts (CFs). In the present study, we extended our investigations to adult rat CFs to explore whether AVPcould induce adult rat CF proliferation and, if so, to identify the mechanism involved. Adult rat CFs were isolated, cultured andsubjected to AVP treatment. DNA synthesis and cell cycle distribution were analyzed by [3H]-thymidine incorporation and flowcytometry. Cellular extracellular signal-regulated kinase 1/2 (ERK1/2) activity was measured by in vitro kinase assay using myelin basicprotein (MBP) as a substrate. Protein expressions of total- and phospho-ERK1/2, p27Kip1, cyclins D1, A, E were assessed by Westernblot. The results showed that AVP stimulated DNA synthesis in adult rat CFs, and the effect was abolished by a V1 receptor antagonist,d(CH2)5[Tyr2(Me), Arg8]-vasopressin (0.1 μmol/L), but not by a V2 receptor antagonist, desglycinamide-[d(CH2)5, D-Ile2, Ile4, Arg8]-vasopressin (0.1 µmol/L). AVP induced an activation of ERK1/2, which could be mimicked by the protein kinase C (PKC) activator,phorbol 12-myristate 13-acetate (PMA, 30 nmol/L, 5 min), but abolished by depletion of PKC via chronic PMA incubation (2.5 μmol/L, 24 h). In addition, AVP down-regulated protein expression of p27Kip1, increased protein expressions of cyclins D1, A and E, andinduced cell cycle progression from G0/G1 into S stage. Inhibition of ERK1/2 activation by PD98059 (30 μmol/L) abolished the effectof AVP on DNA synthesis, protein expressions of p27Kip1, cyclins D1, A and E as well as cell cycle progression. These results suggestthat AVP is also a growth factor for adult rat CFs. The mitogenic effect of AVP is mediated via V1 receptors and PKC-ERK1/2 pathway.Moreover, AVP modulates the expressions of cell cycle regulatory proteins p27Kip1 and cyclins D1, A and E, which lie downstream ofERK1/2 activation, and induces cell cycle progression in adult rat CFs.
Key words: arginine vasopressin; V1 receptor; extracellular signal-regulated kinase 1/2; protein kinase C; p27Kip1; cyclin; cardiacfibroblast
蛋白激酶C-细胞外信号调节激酶1/2通路介导精氨酸升压素对成年大鼠 心肌成纤维细胞的促增殖作用
何燕萍，赵连友*，郑强荪，刘少伟，赵晓燕，陆晓龙，牛晓琳
第四军医大学唐都医院心血管内科，西安 710038
摘 要：精氨酸升压素(arginine vasopressin, AVP)是高血压和心力衰竭时激活的神经体液和血流动力学因子，同时，它还具有直接的生长刺激作用。我们以往的研究显示AVP可诱导新生大鼠心肌成纤维细胞(cardiac fibroblasts, CFs)增殖。本研究旨在进一步观察AVP是否对成年大鼠CFs具有促增殖作用，并探计其机制。采用组织块法培养成年大鼠CFs，用[3H]-TdR掺入法和流式细胞仪方法观察AVP作用下CFs的DNA合成和细胞周期分布。根据特异性底物髓磷脂基质蛋白(myelin basic protein,MBP)的磷酸化水平测定细胞外信号调节激酶1/2 (extracellular signal-regulated kinase 1/2, ERK1/2)的活性。用Western blot检测ERK1/2的磷酸化和p27Kip1、细胞周期蛋白D1、A、E的表达。结果显示，AVP (0.1 μmol/L)可促进成年大鼠CFs的DNAReceived 2008-01-15 Accepted 2008-02-25
This work was supported by the Natural Science Foundation of Shaanxi Province, China (No. 2004C2-21). *Corresponding author. Tel: +86-29-84777656; Fax: +86-29-84777656; E-mail: LianY_Zhao@