姜黄素对TGF-β1诱导的心脏成纤维细胞表达NGF、IL-1β、TNF-α的影响及机制研究

Effect and mechanism of curcumin on expression of nerve growth factor(NGF),interleukin-1β(IL-1β)and tumor necrosis factor-alpha(TNF-α)in cardiac fibroblasts induced by TGF-β1

目的观察姜黄素对转化生长因子-β1(Transforming growth factor-β1,TGF-β1)诱导的心脏成纤维细胞表达神经生长因子(Nerve growth factor,NGF)和炎症因子的影响,并探讨其可能的作用机制。方法在体外通过TGF-β1刺激心脏成纤维细胞,模拟心肌梗死(Myocardial in-farction,MI)模型。分组为Control组、TGF-β1组、美托洛尔(Myto)组,姜黄素微、低、中、高剂量(CCM-5、10、20、40)组。造模时间为48 h,药物处理时间为48 h。使用RT-qPCR检测NGF、Hes1、Hey1、NICD的mRNA表达水平,Western blot法检测NGF、Hes1、Hey1、NICD的蛋白表达水平,Elisa检测细胞上清中炎症因子IL-1β、TNF-α含量。结果与Control组比较,TGF-β1组细胞中NGF、Hes1、Hey1、NICD的mRNA及其蛋白表达水平明显升高(P<0.05),细胞上清中IL-1β、TNF-α含量明显升高(P<0.05)。与TGF-β1组比较,CCM-5组、CCM-10组、CCM-20组、CCM-40组的NGF、Hes1、Hey1、NICD的mRNA及蛋白表达水平与IL-1β、TNF-α含量均有不同程度的降低,部分组间比较差异有统计学意义(P<0.05),并且这些指标具有姜黄素浓度依赖性。结论姜黄素可以下调TGF-β1诱导的心脏成纤维细胞中NGF和IL-1β、TNF-α的表达,具有抗交感神经重构和抗炎作用,其机制可能与Notch信号通路的活化有关。

Objective To observe the effect of curcumin on the expression of nerve growth factor(NGF)and inflammatory factors in cardiac fibroblasts induced by transforming growth factor-β1(TGF-β1),and to explore the possible mechanism of action.Methods Cardiac fibroblasts were stimulated by TGF-β1 in vitro to simulate myocardial infarction(MI)model.They were divided into control group,TGF-β1 group,metoprolol(Myto)group and curcumin small,low,medium and high dose(CCM 5,10,20,40)groups.The modeling time was 48 hours,and the drug treatment time was 48 hours.The mRNA expression levels of NGF,Hes1,Hey1 and NICD were detected by RT-qPCR,the protein expression levels of NGF,Hes1,Hey1 and NICD were detected by western blot,and the contents of inflammatory factors IL-1βand TNF-αin the cell supernatant were detected by Elisa.Results Compared with control group,the expression lev-els of NGF,Hes1,Hey1 and NICD mRNA and their proteins in TGF-β1 group were significantly increased(P<0.05),the contents of IL-1βand TNF-αin cell supernatant were significantly increased(P<0.05).Compared with TGF-β1 group,the expression levels of NGF,Hes1,Hey1 and NICD mRNA and their proteins in CCM-5 group,CCM-10 group,CCM-20 group and CCM-40 group were decreased to varying degrees,as well as the contents of IL-1βand TNF-α.The comparison among the groups were statistically significant(P<0.05).These indicators were concentration dependent on curcumin.Conclusion Curcu-min can down-regulate the expression of NGF,IL-1βand TNF-αin cardiac fibroblasts induced by TGF-β1,and it has anti-sympathetic remodeling and anti-inflammatory effects,which may be related to the activa-tion of Notch signaling pathway.