Apelin抑制小鼠气道平滑肌细胞增殖的作用及机制  

Inhibitive role of apelin in aiway smooth muscle cells proliferation in mice and its potential mechanisms

作　　者：吴媛媛[1] 孙秀珍[1] 刘昀[1] 陈海娟[1] WU Yuanyuan;SUN Xiuzhen;LIU Yun;CHEN Haijuan(The Second Affiliated Hospital of Medical College,Xi'an Jiaotong University,Xi'an 710004,China)

机构地区：[1]西安交通大学第二附属医院,西安710004

出　　处：《实用医学杂志》2019年第2期177-180,共4页The Journal of Practical Medicine

基　　金：西安交通大学第二附属医院科研基金(编号:YJ(QN)201522);国家自然科学基金面上项目(编号:81770037)

摘　　要：目的探讨apelin抑制PDGF诱导小鼠气道平滑肌细胞(ASMCs)增殖的作用及其分子机制。方法采用组织块贴壁法培养原代BALB/c小鼠ASMCs,以PDGF(20 ng/mL)诱导ASMCs增殖建立模型;将ASMCs分为3组:正常对照组、PDGF刺激组、PDGF+apelin干预组,应用BrdU试剂盒检测各组细胞的增殖效应,应用Western blot检测各组中TGF-β1、p-Smad3和p27蛋白水平;沉默APJ基因,将ASMCs分为4组:正常对照组、PDGF刺激组、PDGF+apelin干预组、APJ shRNA+apelin+PDGF干预组,再次检测各组ASMCs的增殖情况,以及TGF-β1、p-Smad3和p27蛋白水平。结果 Apelin(1 nmol/L)可以有效抑制PDGF刺激ASMCs增殖的作用(P <0.05);apelin可以抑制PDGF升高TGF-β1、p-Smad3水平、下调p27表达的作用;转染APJ shRNA,可以逆转apelin抑制ASMCs增殖,下调TGF-β1、p-Smad3,增加p27蛋白水平的作用。结论 Apelin与其受体APJ结合,可有效抑制PDGF刺激的ASMCs增殖,其机制可能是通过抑制TGF-β/Smad信号通路而实现。Objective To investigate the effect of apelin on PDGF-induced ASMCs proliferation and its underlying mechanisms.Methods ASMCs were isolated from the airway of BALB/c mice using tissue adherent culture method,and then a cell proliferation model in vitro induced by PDGF(20 ng/mL)was established.ASMCs were divided into 3 groups according to the different interference:control group,PDGF stimulation group and PDGF+apelin group.The BrdU ELISA Kit was used to examine ASMCs proliferation and western blotting to detect TGF-β1,p27 and p-Smad3 protein levels.Next,ASMCs were transfected with sequence-specific APJ siRNA,and then ASMCs were divided into four groups:control group,PDGF stimulation group,apelin+PDGF group and shRNA APJ+apelin+PDGF group.ASMCs proliferation and TGF-β1,p27 and p-Smad3 protein levels were in vestigated again using the same methods above.Results Pretreatment with apelin(1 nmol/L)inhibited PDGF-in duced ASMCs proliferation(P<0.05),reduced TGF-β1 and p-Smad3 level,but raised p27 protein level.Knock down of APJ by transfection APJ shRNA could reverse the effect of PDGF on ASMCs proliferation,increase the lev el of TGF-β1 and p-Smad3 and decrease the expression of p27.Conclusion Apelin/APJ negatively regulates PDGF-induced ASMCs proliferation and the underling mechanism may be the inhibition of TGF-β/Smad signaling pathway.

关 键 词：支气管哮喘 APELIN 细胞增殖 TGF-Β/SMAD信号通路 

分 类 号：R562.25[医药卫生—呼吸系统]