尾加压素Ⅱ通过ERK1/2途径诱导大鼠肺动脉平滑肌细胞增殖和上调Egr-1表达  

Urotensin Ⅱinduces pulmonary arterial smooth muscle cell proliferation and up-regulates Egr-1 expression through ERK1 /2 pathway in rats

作　　者：孟庆红[1,2] 蔡直峰 赵翠芬[1] 李福海[1] 孔清玉[1] 夏伟[1] 李栋[3] 

机构地区：[1]山东大学齐鲁医院儿科,山东济南250012 [2]潍坊市人民医院新生儿科,山东潍坊261041 [3]山东大学齐鲁医院低温医学研究室,山东济南250012

出　　处：《中国病理生理杂志》2014年第11期2093-2097,共5页Chinese Journal of Pathophysiology

基　　金：山东省自然科学基金资助项目(No.Y2008C44)

摘　　要：目的:研究不同浓度尾加压素Ⅱ(UⅡ)及其受体拮抗剂urantide对培养大鼠肺动脉平滑肌细胞(PASMC)增殖的影响,探讨丝裂原活化蛋白激酶(MAPK)途径及早期生长反应因子1(Egr-1)在UⅡ诱导大鼠PASMCs增殖中的作用。方法:以组织贴块法原代培养大鼠PASMCs:(1)采用Brd U掺入实验测定不同浓度(1μmol/L、0.1μmol/L和0.01μmol/L)UⅡ及其受体拮抗剂对大鼠PASMCs增殖的影响;(2)采用real-time PCR检测UⅡ及其受体拮抗剂对细胞外信号调节激酶1/2(ERK1/2)、应激活化蛋白激酶(SAPK)、p38 MAPK及Egr-1mRNA表达的影响;(3)采用Western blotting检测UⅡ、urantide及ERK1/2抑制剂PD98059对PASMCs磷酸化ERK1/2(p-ERK1/2)、p-SAPK、p-p38 MAPK和Egr-1蛋白表达的影响。结果:(1)UⅡ在一定浓度范围(1μmol/L、0.1μmol/L和0.01μmol/L)呈浓度依赖性地促进肺动脉平滑肌细胞增殖(P<0.01或P<0.05),这种增殖作用可被urantide拮抗(P<0.05);(2)UⅡ上调ERK1/2、SAPK及Egr-1 mRNA表达(P<0.01或P<0.05),PD98059和(或)urantide可拮抗UⅡ诱导的ERK1/2、SAPK和Egr-1 mRNA的表达(P<0.01或P<0.05),但对p-p38 MAPK无影响;(3)UⅡ可增加PASMCs p-ERK1/2、p-SAPK及Egr-1蛋白表达(P<0.01或P<0.05),urantide及PD98059可拮抗UⅡ诱导的p-ERK1/2、p-SAPK和Egr-1蛋白表达(P<0.01,P<0.05)。结论:UⅡ可能通过ERK1/2途径诱导PASMCs增殖和上调Egr-1表达,Egr-1可能通过激活ERK1/2途径参与了PASMCs的增殖。AIM： To investigate the effect of urotensinⅡ（ UⅡ） on the proliferation of cultured rat pulmonary arterial smooth muscle cells（ PASMCs）,and to explore whether mitogen-activated protein kinase（ MAPK） signaling pathways and early growth response factor-1（ Egr-1） involved in the regulation of the PASMCs proliferation stimulated by UⅡ.METHODS： The rat PASMCs were isolated and cultured in vitro with explant culture technique. The proliferation of cultured PASMCs stimulated by different doses of UⅡ was detected by Brd U incorporation. The mRNA expression of extracellular signal-regulated kinase 1 /2（ ERK1 /2）,stress-activated protein kinase（ SAPK）,p38 MAPK and Egr-1 in cultured PASMCs treated with UⅡ,U Ⅱ-specific antagonist urantide,and ERK1 /2 inhibitor PD98059 was detected by real-time PCR. The protein levels of phosphorylated ERK1 /2（ p-ERK1 /2）,p-SAPK,p-p38 and Egr-1 in cultured PASMCs were determined by Western blotting. RESULTS： UⅡ at concentrations of 1 μmol / L,0. 1 μmol / L and 0. 01 μmol / L increased the proliferation of cultured PASMCs in a dose-dependent manner（ P〈0. 01 or P〈0. 05）,with the maximal effect at a concentration of 1 μmol / L. However,urantide inhibited the promotion effect of UⅡ on PASMC proliferation（ P〈0. 05）.UⅡ up-regulated the mRNA expression of ERK1 /2,SAPK and Egr-1（ P〈0. 01 or P〈0. 05）,but not the p38 MAPK.However,the up-regulatory effect of U Ⅱ on ERK1 /2 and Egr-1 expression was inhibited by PD98059 and / or urantide（ P〈0. 01 or P〈0. 05）. UⅡ also increased the protein levels of p-ERK1 /2,p-SAPK and Egr-1（ P〈0. 01 or P〈0. 05）,but the promotion effect was also inhibited by PD98059 and / or urantide（ P〈0. 01 or P〈0. 05）. CONCLUSION： UⅡ increases the proliferation of PASMCs,and U Ⅱand Egr-1 participates in UⅡ-mediated proliferation of cultured PASMCs through activation of ERK1 /2 signal pathway.

关 键 词：尾加压素Ⅱ 肺动脉平滑肌细胞 丝裂原活化蛋白激酶 早期生长反应因子-1 肺血管重构 

分 类 号：R363.2[医药卫生—病理学]