中文摘要：

目的 探讨糖基化终产物（AGE）对培养乳鼠心肌成纤维细胞增殖、结缔组织生长因子（CTGF）和Smad2、Smad4蛋白表达的影响及罗格列酮的干预作用.方法 采用胰酶消化法和差速贴壁分离法获取心肌成纤维细胞,应用MTT法、流式细胞仪法分别观察不同浓度AGE及罗格列酮对心肌成纤维细胞的细胞增殖、细胞周期的影响,ELISA方法 检测细胞培养上清中TGF-β1水平,Westem印迹技术检测CTGF及Smad2、Smad4蛋白质表达.结果 在一定浓度范围内,AGE干预心肌成纤维细胞,随浓度的增加,细胞增殖更加显著,TGF-β1分泌增加,CTGF蛋白表达增加.罗格列酮（0.1、1、10μmoVL）干预后,随浓度的增加,抑制心肌成纤维细胞增殖（分别为0.823±0.072、0.785±0.060、0.601±0.081对0.981±0.049,P〈0.05）、抑制心肌成纤维细胞分泌TGF-β1（分别为257.77±9.09、230.29±6.56、200.84±10.26对300,68±8.56,P〈0.01）、抑制CTGF蛋白表达的作用都更加显著（分别为0.769±O.108、0.590±0.095、0.534±0.115对1.021±0.113,P〈0.01）.罗格列酮（1和10μmol/L）干预后可显著减少AGE诱导的Smad2的蛋白表达（分别为0.424±0.059对0.572±0.073,P〈0.05;0.396±0.080对0.572±0.073,P〈0.01）,同时可显著减少AGE诱导的Smad4的蛋白表达（分别为0.580±0.063对0.672±0.059,P〈0.05;0.556±0.051对0.672±0.059,P〈0.01）.结论 AGE刺激心肌成纤维细胞增殖及分泌TGF-β1,同时诱导CTGF、Smad2及Smad4蛋白表达,罗格列酮在一定程度上抑制AGE上述作用,表明罗格列酮抑制心肌成纤维细胞增殖的效应与CTGF/Smad通路密切相关.

英文摘要：

Objective To investigate the effects of rosiglitazone on the proliferation,connective tissue growth factor and Smad expression in cultured cardiac fibroblasts induced by advanced glycosylation end-products （AGEs）.Methods After being treated with various amounts of rosiglitazone,the cultured neonatal rat cardiac fibroblasts were incubated with AGEs.The status of cardiac fibroblasts proliferation and cell cycle were detected by 3-（4,5-dimethyhhiazol-2-yl） -2,5-diphenyl tetrazolium bromide （MTI） assay and flow cytometry.Furthermore,ELISA technique was applied to identify the level of TGF-β1.The protein expressions of CTGF and Smad in cardiac fibroblasts of neonatal SD rats were detected with Western blotting.Results The exposure of cardiac fibroblasts to AGEs at doses of 0-200 mg/L induced a dose-dependent increase in cell proliferation.At the concentration of rosiglitazooe （0.1,1,and 10 μmol/L）,the cell proliferation was reduced compared with 200 mg/L AGEs group by O.823±0.072,0.785±0.060,0.601±0.081 vs 0.981±0.049,respectively （P 〈 0.05）.The increased levels of TGF-β1 in supematants of cultured cardiac fibroblasts stimulated by AGEs were inhibited by rosiglitazone at the concentrations of 0.1,1,10μmol/L by 257.77±9.09,230.29±6.56,200.84±10.26 vs 300.68±8.56,respectively （vs 200 mg/L AGEs,P〈0.01）.Western blot indicated that pretreatment with rosiglitazone （0.1,1,and 10 μmol/L） inhibited CTGF protein production in a dose-dependent by 0.769±0.108,0.590±0.095,0.534±0.115 vs 1.021±0.113,respectively （vs 200 mg/L AGEs,P〈0.01）.It was also demonstrated that pretreatment with rosiglitazone （1 and 10 μmol/L） inhibited Smad2 protein production by 0.424±0.059,0.396±O.080 vs 0.572±0.073,respectively （vs 200 mg/L AGEs,P 〈 0.05 or P 〈 0.01）.Meanwhile pretreatment with rosiglitazone （1 and 10 μmol/L） inhibited Smad4 protein production by 0.580±0.063,0.556±0.051 vs 0.672±0.059,respectively （vs 200 mg/L AGEs,P 〈 0.05 or P 〈 0.01）.Conclusions The