Yes相关蛋白调控肺动脉平滑肌细胞增殖的分子机制  

Molecular mechanisms of Yes-associated protein in regulating the proliferation of pulmonary arterial smooth muscle cells

作　　者：柯蕊 和平[1] 张伟[1] 史文花 张永红[1] 刘原[2] Ke Rui;He Ping;Zhang Wei;Shi Wenhua;Zhang Yonghong;Liu Yuan(Department of Respiratory and Critical Care Medicine,the Second Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710004,China;Department of Health Examination,the Second Affiliated Hospital of 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呼吸与危重症医学科,西安710004 [2]西安交通大学第二附属医院健康体检部,西安710004

出　　处：《国际呼吸杂志》2024年第1期73-78,共6页International Journal of Respiration

基　　金：国家自然科学基金(82100067)。

摘　　要：目的研究Yes相关蛋白(YAP)调控肺动脉平滑肌细胞(PASMCs)增殖的分子机制。方法本研究为实验研究。选取健康SD大鼠,体质量70~80 g。培养大鼠PASMCs,根据是否给予S1P刺激将PASMCs分为S1P组和对照组,蛋白质印迹法检测细胞磷酸化YAP(p-YAP)和β-catenin、CyclinD1蛋白水平。按照转染control干扰小RNA(siRNA)或YAP siRNA后再给予S1P刺激的情况不同,将PASMCs分为对照组、S1P组、si+S1P组、si-YAP+S1P组,蛋白质印迹法检测各组细胞中β-catenin和CyclinD1的蛋白水平。按照转染control siRNA或β-catenin siRNA后再给予S1P刺激的情况不同,将PASMCs分为对照组、S1P组、si+S1P组、si-β-catenin+S1P组,蛋白质印迹法检测各组细胞中CyclinD1的蛋白水平。按照转染control siRNA或YAP siRNA或β-catenin siRNA后再给予S1P刺激的情况不同,将PASMCs分为对照组、S1P组、si+S1P组、si-YAP+S1P组和si-β-catenin+S1P组,BrdU掺入法检测各组细胞增殖情况。结果S1P组p-YAP水平低于对照组[(0.61±0.09)比(1.00±0.11),P=0.009],β-catenin和CyclinD1蛋白水平高于对照组[(1.98±0.14)比(1.00±0.10),(1.94±0.15)比(1.00±0.08),均P=0.001]。si-YAP+S1P组β-catenin、CyclinD1水平均低于S1P组[(1.15±0.09)比(1.95±0.12),(1.18±0.16)比(1.96±0.05),均P<0.01]。si-β-catenin+S1P组CyclinD1水平低于S1P组[(1.18±0.24)比(1.95±0.24),P<0.01]。S1P组细胞增殖率高于对照组[(169.69±12.85)%比(100.00±12.52)%,P<0.01],si-YAP+S1P组、si-β-catenin+S1P组细胞增殖率均低于S1P组[(126.33±14.56)%比(169.69±12.85)%,(128.10±15.25)%比(169.69±12.85)%,均P<0.01]。结论YAP/β-catenin/CyclinD1信号通路可以调控PASMCs增殖。ObjectiveTo examine the molecular mechanisms of Yes-associated protein(YAP)in regulating the proliferation of pulmonary arterial smooth muscle cells(PASMCs).MethodsIt was an experimental study.PASMCs were separated from healthy Sprague-Dawley(SD)rats weighing 70-80 g.Primary PASMCs were cultured and stimulated with S1P or blank control,followed by the detection of the expressions of phosphorylated YAP(p-YAP),β-catenin and CyclinD1 by Western blot.Then,PASMCs were induced with blank control,S1P,S1P+transfection of si-NC,and S1P+transfection of si-YAP,followed by the detection of the protein expressions ofβ-catenin and CyclinD1 by Western blot and assessment of cell proliferation by BrdU assay.PASMCs were further induced with blank control,S1P,S1P+transfection of si-NC,and S1P+transfection of si-β-catenin,followed by the detection of the protein expression of CyclinD1 by Western blot and assessment of cell proliferation by BrdU assay.ResultsThe expression of p-YAP([0.61±0.09]vs[1.00±0.11],P=0.009)was significantly lower in S1P-induced PASMCs than that of blank control,while protein expressions ofβ-catenin([1.98±0.14]vs[1.00±0.10],P=0.001)and CyclinD1([1.94±0.15]vs[1.00±0.08],P=0.001)were significantly higher.Protein expressions ofβ-catenin([1.15±0.09]vs[1.95±0.12],P<0.01)and CyclinD1([1.18±0.16]vs[1.96±0.05],P<0.01)were significantly lower in PASMCs induced with S1P+transfection of si-YAP than those induced with S1P.Protein expression of CyclinD1([1.18±0.24]vs[1.95±0.24],P<0.01)was significantly lower in PASMCs induced with S1P+transfection of si-β-catenin than those induced with S1P.The proliferative rate was significantly higher in PASMCs induced with S1P([169.69±12.85]%vs[100.00±12.52]%,P<0.01)than that of blank control.The proliferative rate was significantly lower in PASMCs induced with S1P+transfection of si-YAP([126.33±14.56]%vs[169.69±12.85]%,P<0.01)and S1P+transfection of si-β-catenin([128.10±15.25]%vs[169.69±12.85]%,P<0.01)than that induced with S1P.ConclusionsThe YAP/β-catenin/C

关 键 词：肺动脉高压 肺动脉平滑肌细胞 细胞增殖 Yes相关蛋白 Β-CATENIN 

分 类 号：R544.1[医药卫生—心血管疾病]