芦丁对TGF-β1诱导的心肌成纤维细胞纤维化的影响  

Effect of rutin on TGF-β1 induced fibrosis of cardiac fibroblasts

作　　者：张冠楠 牛丕莲 井瑞欣 石新卫 白明生[1] ZHANG Guannan;NIU Pilian;JING Ruixin;SHI Xinwei;BAI Mingsheng(School of Life Sciences,Ningxia University,Yinchuan 750021,China;Shanxi Institute of Botany,Xi’an 710061,China)

机构地区：[1]宁夏大学生命科学学院,银川750021 [2]陕西植物研究所,西安710061

出　　处：《天津中医药》2025年第1期71-77,共7页Tianjin Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金地区科学基金项目(31860583)。

摘　　要：[目的]探究芦丁对转化生长因子-β1(TGF-β1)诱导的心肌成纤维细胞(CFs)纤维化及TGF-β1/Smad信号通路的影响。[方法]构建TGF-β1诱导的CFs细胞模型,设置组别为:对照组(完全培养基+细胞)、模型组(含10 ng/mL TGF-β1+细胞)、阳性对照组(10 ng/mL TGF-β1+10μmol/L卡托普利和细胞)和给药组(10 ng/mL TGF-β1+200μg/mL的芦丁和细胞)。采用蛋白免疫印迹法检测α-平滑肌肌动蛋白(α-SMA)、波形蛋白(Vim)、纤连蛋白(FN)、胶原蛋白Ⅰ(CollageⅠ)、胶原蛋白Ⅲ(CollageⅢ)、TGF-β1/Smad通路相关蛋白的表达情况;采用实时荧光定量聚合酶链式反应(qPCR)法检测Acta 2、Vim、FN、ColⅠ1a1、基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)的基因表达水平;采用免疫荧光技术检测关键蛋白α-SMA的表达及定位来探究芦丁对TGF-β1诱导的CFs纤维化的影响。[结果]用TGF-β1诱导CFs后,成功激活Vim、FN、CollageⅠ、CollageⅢ、TGF-β1/Smad信号通路相关蛋白的表达,200μg/mL芦丁干预后,抑制了Vim、FN、CollageⅠ、CollageⅢ、TGF-β1/Smad信号通路相关蛋白和mRNA的激活从而缓解了心肌纤维化的发生;免疫荧光实验结果显示:与对照组相比,TGF-β1模型组中α-SMA绿色荧光强度显著增强;200μg/mL芦丁组与模型组相比,α-SMA绿色荧光强度明显减弱。[结论]本研究结果表明200μg/mL芦丁通过抑制TGF-β1诱导的Vim、FN、CollageⅠ、CollageⅢ、α-SMA以及TGF-β1/Smad信号通路相关蛋白和mRNA的表达,从而缓解心肌纤维化。[Objective]To explore the effect of rutin on transforming growth factor-β1(TGF-β1)-induced fibrosis of cardiac fibroblasts(CFs)and the TGF-β1/Smad signaling pathway.[Methods]The CFs cell model induced by TGF-β1 was constructed,and the groups were set as follows:control group(complete medium+cells),model group(containing 10 ng/mL TGF-β1+cells),positive control group(10 ng/mL TGF-β1+10μmol/L captopril and cells)and administration group(10 ng/mL TGF-β1+200μg/mL rutin and cells).The expression ofα-smooth muscle actin(α-SMA),Vimentin(Vim),Fibronectin(FN),Collagen I,CollagenⅢand TGF-β1/Smad pathway-related proteins were detected by Western blot.The gene expression levels of Acta 2,Vim,FN,ColⅠ1a1,matrix metalloproteinase 2(MMP-2)and matrix metalloproteinase 9(MMP-9)were detected by quantitative real-time polymerase chain reaction(qPCR).Immunofluorescence was used to detect the expression and localization of the key proteinα-SMA to explore the effect of rutin on TGF-β1-induced CFs fibrosis.[Results]After CFs were induced by TGF-β1,the expression of Vim,FN,Collage I,CollageⅢ,TGF-β1/Smad signaling pathway-related proteins was successfully activated.After intervention with 200μg/mL rutin,the activation of Vim,FN,CollageⅠ,CollageⅢ,TGF-β1/Smad signaling pathway-related proteins and mRNA was inhibited,thereby alleviating the occurrence of myocardial fibrosis.The results of immunofluorescence assay showed that compared with the control group,the green fluorescence intensity ofα-SMA in the TGF-β1 model group was significantly enhanced.Compared with the model group,the green fluorescence intensity ofα-SMA was significantly weakened in the 200μg/mL rutin group.[Conclusion]The results of this study showed that 200μg/mL rutin alleviated myocardial fibrosis by inhibiting the expression of Vim,FN,Collage I,CollageⅢ,α-SMA and TGF-β1/Smad signaling pathway-related proteins and mRNA induced by TGF-β1.

关 键 词：芦丁 心肌纤维化 心肌成纤维细胞 TGF-Β1/SMAD信号通路 

分 类 号：R542.23[医药卫生—心血管疾病]