摘要
目的:探讨莱菔硫烷(sulforaphane,SFN)对转化生长因子β1(TGF-β1)诱导的大鼠心肌纤维化的影响及其机制。方法:用TGF-β1处理大鼠心肌成纤维细胞构建心肌纤维化模型;用SFN(5、10、20μg/ml)处理TGF-β1诱导的成纤维细胞;采用CCK8、Transwell法检测细胞的活性、迁移,采用Western blot法检测细胞中基质金属蛋白酶9(MMP-9)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、Ⅰ型胶原蛋白(ColⅠ)、ColⅢ、TGF-β1、p-Smad3、p-Smad2、Smad2/3的蛋白表达;采用酶联免疫吸附试验(ELISA)法检测细胞上清液中活性氧(ROS)、丙二醛(MDA)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)的含量。结果:与对照组比较,TGF-β1组细胞增殖和迁移能力均显著升高(P<0.05),并且MMP-9、α-SMA、ColⅠ、ColⅢ、TGF-β1、Smad2/3的蛋白表达均显著升高(P<0.05),p-Smad3、p-Smad2的蛋白表达显著降低(P<0.05),ROS、MDA的含量显著升高(P<0.05),GSH、SOD的含量显著降低(P<0.05);与TGF-β1组比较,SFN(5、10、20μg/ml)处理后,细胞的上述变化均显著减轻。结论:SFN可抑制TGF-β1诱导的大鼠心肌成纤维细胞的异常增殖、迁移、氧化应激和纤维化,其机制与调节TGF-β1/Smads信号通路的活性有关。
Objective:To investigate the effect of sulforaphane(SFN)on myocardial fibrosis induced by TGF-β1 in rats and its mechanism.Methods:TGF-β1 was used to establish myocardial fibrosis model.The cardiac fibroblasts were treated with SFN(5,10,20μg/ml).CCK8 and Transwell assays were performed to detect the cell activity and migration.The protein expressions ofα-smooth muscle actin(α-SMA),ColⅠ,ColⅢ,TGF-β1,p-smad3,p-Smad2 ans Smad2/3 were detected by Western blot.The contents of reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH)and superoxide dismutase(SOD)in cells were detected by ELISA.Results:Compared with the control group,the cell activity and migration ability significantly increased in TGF-β1 group(P<0.05).The protein expressions ofα-SMA,ColⅠ,ColⅢ,TGF-β1 and Smad2/3 significantly increased,while the protein levels of p-Smad3 and p-Smad2 significantly decreased in TGF-β1 group(P<0.05).And the contents of ROS and MDA significantly increased(P<0.05),while the contents of GSH and SOD significantly decreased in TGF-β1 group(P<0.05).Compared with TGF-β1 group,the above indexes were significantly reduced in SFN(5,10,20μg/ml)groups.Conclusion:SFN can inhibit the abnormal proliferation,migration,oxidative stress and fibrosis of cardiac fibroblasts induced by TGF-β1,which may be related to the regulation of TGF-β1/Smads signaling pathway.
作者
张莉
张磊
王艳梅
章辰琛
王鑫
ZHANG Li;ZHANG Lei;WANG Yanmei;ZHANG Chenchen;WANG Xin(Department of Clinical Medicine,West Anhui Health Vocational College,Liuan 237005,China)
出处
《沈阳医学院学报》
2023年第3期234-237,251,共5页
Journal of Shenyang Medical College
基金
安徽省卫生厅医药科研计划项目(No.12925KJ2017B41)。
