白藜芦醇调控肺动脉平滑肌细胞增殖的作用和分子机制  

Effect of resveratrol in regulating the proliferation of pulmonary arterial smooth muscle cells and the underlying molecular mechanism

作　　者：史文花 刘娟[1] 张永红[1] 柯蕊 Shi Wenhua;Liu Juan;Zhang Yonghong;Ke Rui(Department of Respiratory and Critical Care Medicine,the Second Affiliated Hospital of Xi'an Jiaotong University,Xian 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

出　　处：《国际呼吸杂志》2024年第7期825-830,共6页International Journal of Respiration

基　　金：国家自然科学基金(82100067)。

摘　　要：目的探讨白藜芦醇(Res)调控肺动脉平滑肌细胞(PASMCs)增殖的作用和分子机制。方法本研究为实验研究。体外分离培养大鼠原代PASMCs,采用免疫荧光法鉴定PASMCs,选用第3~6代细胞进行后续实验。比较不同浓度Res(10μmol/L Res干预组、30μmol/L Res干预组、100μmol/L Res干预组和300μmol/L Res干预组)对半乳糖凝集素3(Gal-3)诱导PASMCs增殖的影响,确定Res的最终干预浓度。根据给予Gal-3、Res、TD139处理和转染control干扰小RNA(siRNA)、Yes相关蛋白(YAP)siRNA的情况不同,将PASMCs分为7组:对照组(无处理)、Gal-3刺激组(30 nmol/L Gal-3刺激48 h)、Res刺激组(100μmol/L Res刺激48 h)、Res干预组(100μmol/L Res处理1 h,再加入30 nmol/L Gal-3刺激48 h)、TD139干预组(10μmol/L TD139作用1 h,再加入100μmol/L Res干预1 h和30 nmol/L Gal-3刺激48 h)、si-control组(转染control siRNA 24 h,再加入100μmol/L Res干预1 h和30 nmol/L Gal-3刺激48 h)和si-YAP组(转染YAP siRNA 24 h,再加入100μmol/L Res干预1 h和30 nmol/L Gal-3刺激48 h)。采用CCK-8实验检测PASMCs增殖能力。采用蛋白质印迹法检测对照组、Res刺激组PASMCs中Gal-3和YAP的表达水平,以及对照组、Gal-3刺激组、Res干预组和TD139干预组PASMCs中YAP的表达水平。结果Gal-3刺激组细胞增殖能力高于对照组,100μmol/L Res干预组、300μmol/L Res干预组细胞增殖能力均低于Gal-3刺激组[(1.36±0.10)比(0.87±0.13),(1.11±0.09)比(1.36±0.10),(1.00±0.11)比(1.36±0.10),均P<0.05],后续实验中将100μmol/L作为Res的干预浓度。Res刺激组Gal-3和YAP的表达水平均低于对照组[(0.49±0.12)比(1.00±0.10),(0.62±0.13)比(1.00±0.09),均P<0.05]。Gal-3刺激组YAP表达水平高于对照组,Res干预组YAP表达水平低于Gal-3刺激组,TD139干预组YAP表达水平低于Res干预组[(1.84±0.09)比(1.00±0.10),(1.13±0.11)比(1.84±0.09),(0.82±0.15)比(1.13±0.11),均P<0.05]。Gal-3刺激组细胞增殖能力高于对照组,Res干预组细胞增殖能力低�Objective:To explore the effect of resveratrol(Res)in regulating the proliferation of pulmonary arterial smooth muscle cells(PASMCs)and the underlying molecular mechanism.Methods:This was an experimental study.Primary rat PASMCs were isolated and cultured in vitro,and verified by immunostaining.The 3rd to 6th generation cells were used in the following experiments.The effects of Res at varying concentrations(10,30,100 and 300μmol/L)on the proliferation of Galectin-3(Gal-3)-induced PASMCs were evaluated,and the final intervention concentration of Res was determined.PASMCs were divided into 7 groups based on the treatment with Gal-3,Res,and TD139,as well as the transfection of control small interfering RNA(siRNA)and Yes-associated protein(YAP)siRNA as follows:control group(blank control),Gal-3 stimulation group(30 nmol/L Gal-3 for 48 h),Res stimulation group(100μmol/L Res for 48 h),Res intervention group(100μmol/L Res for 1 h+30 nmol/L Gal-3 for 48 h),TD139 intervention group(10μmol/L TD139 for 1 h+100μmol/L Res for 1 h+30 nmol/L Gal-3 for 48 h),si-control group(transfection of si-control for 24 h+100μmol/L Res for 1 h+30 nmol/L Gal-3 for 48 h)and si-YAP group(si-YAP for 24 h+100μmol/L Res for 1 h+30 nmol/L Gal-3 for 48 h).Cell proliferation was detected by CCK-8 assay.Expression levels of Gal-3 and YAP in PASMCs of the control group and Res stimulation group,and YAP level in the control group,Gal-3 stimulation group,Res intervention group and TD139 intervention group were detected by Western blot.Results:The proliferative rate was significantly higher in PASMCs of Gal-3 stimulation group than that of control group([1.36±0.10]vs[0.87±0.13],P<0.05),while that was significantly lower in PASMCs treated with 100μmol/L Res([1.11±0.09]vs[1.36±0.10],P<0.05)or 300μmol/L Res([1.00±0.11]vs[1.36±0.10],P<0.05)than Gal-3 stimulation group.In the following experiments,100μmol/L Res was adopted as the optimal concentration.Expression levels of Gal-3([0.49±0.12]vs[1.00±0.10],P<0.05)and YAP([0.62±0.13]vs[1.00±0.

关 键 词：肺动脉高压 肺动脉平滑肌细胞 细胞增殖 白藜芦醇 半乳糖凝集素3 Yes相关蛋白 

分 类 号：R285[医药卫生—中药学]