FGF21对心脏成纤维细胞增殖、胶原合成的影响及在风心病房颤患者心肌中的表达研究
发布时间：2018-08-18 13:44
【摘要】：第一部分FGF21对TGF-β1诱导的心脏成纤维细胞增殖的影响目的探讨成纤维细胞生长因子FGF21对转化生长因子β1 (TGF-β1)诱导的心脏成纤维细胞增殖转化的影响。方法本部分实验分为空白对照组、TGF-β1组、FGF21组、TGF-β1+FGF21组,不同刺激因素干预心脏成纤维细胞至预定时间后,CCK8试剂盒检测每组心脏成纤维细胞的增殖情况,细胞免疫荧光染色方法观察每组心脏肌成纤维细胞标志物α-SMA的表达量,实时荧光定量PCR技术检测细胞增殖标志物PCNA、Ki67的mRNA含量。结果与空白对照组相比,TGF-β1可显著增加心脏成纤维细胞的增殖速率,细胞增殖标志物PCNA, Ki67的mRNA含量显著提高；且TGF-β1可显著增加心脏肌成纤维细胞标志物α-SMA的表达量。而FGF21可显著抑制TGF-β1诱导的心脏成纤维细胞增殖、以及抑制心脏成纤维细胞向心脏肌成纤维细胞的转化。结论成纤维细胞生长因子FGF21可抑制TGF-β1诱导的心脏成纤维细胞增殖转化。第二部分FGF21对TGF-β1诱导的心脏成纤维细胞胶原蛋白合成的影响目的探讨成纤维细胞生长因子FGF21对转化生长因子-β1 (TGF-β1)诱导的心脏成纤维细胞胶原蛋白合成的影响。方法本部分实验分为空白对照组、TGF-β1组、TGF-β1+FGF21组,不同刺激因素干预心脏成纤维细胞至预定时间后,实时荧光定量PCR技术检测心肌纤维化标志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量,此外,蛋白免疫印迹法检测Collagen Ⅰ和Collagen Ⅲ的蛋白表达水平。结果与空白对照组相比,TGF-β1可显著增加心肌纤维化标志物CTGF、CollagenI和Collagen Ⅲ的mRNA含量；且Collagen Ⅰ和Collagen Ⅲ的蛋白表达水平也显著提高。而FGF21可显著抑制心肌纤维化标志物CTGF、Collagen Ⅰ和CollagenⅢ的mRNA含量、以及抑制Collagen Ⅰ和Collagen Ⅲ的蛋白表达水平。结论成纤维细胞生长因子FGF21可抑制TGF-β.诱导的心脏成纤维细胞胶原蛋白合成。第三部分FGF21在风湿性心脏病心房颤动患者心肌组织中的表达情况目的检测风湿性心脏病心房颤动患者血清和心房肌组织中成纤维细胞生长因子FGF21的蛋白水平,探讨FGF21与风湿性心脏病心房颤动患者心房纤维化的关系。方法在换瓣术中收集风湿性心脏病患者的血清和右心房肌组织,分为窦性心律组和心房颤动组。HE病理切片染色观察每组心肌细胞横截面积,Masson染色及心肌纤维化标志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量评估心房纤维化；此外,ELISA试剂盒检测每组患者血清的FGF21水平,免疫组织化学染色及实时荧光定量PCR技术检测心房肌组织中FGF21的表达水平。最后,线性相关分析法评估风湿性心脏病患者FGF21表达水平与心房纤维化严重程度的联系。结果与窦性心律组相比较,心房颤动组患者的心肌细胞横截面积、胶原容积分数、心肌纤维化标志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量均显著增加。此外,心房颤动患者血清FGF21水平显著升高,免疫组织化学染色显示心房颤动患者心肌组织FGF21分布量显著增高、且FGF21的mRNA含量也显著升高。线性相关分析发现FGF21的表达水平与心房纤维化程度呈正相关。结论风湿性心脏病心房颤动患者血清及心肌组织中FGF21的表达显著增加,与心房纤维化严重程度密切相关,参与心房颤动的发生与维持,FGF21或许可成为风湿性心脏病心房颤动患者心房纤维化的生物标志物之一。
[Abstract]:Part I Effects of FGF21 on cardiac fibroblasts proliferation induced by TGF-beta 1 Objective To investigate the effect of fibroblast growth factor-21 on cardiac fibroblasts proliferation and transformation induced by transforming growth factor-beta 1 (TGF-beta 1). CCK8 kit was used to detect the proliferation of cardiac fibroblasts. The expression of myofibroblast marker alpha-SMA was detected by immunofluorescence staining. The expression of PCNA and Ki67 mRNA was detected by real-time quantitative PCR. Compared with the control group, TGF-beta 1 significantly increased the proliferation rate of cardiac fibroblasts, the mRNA content of cell proliferation markers PCNA and Ki67 was significantly increased, and TGF-beta 1 significantly increased the expression of cardiac fibroblasts marker alpha-SMA. Conclusion Fibroblast growth factor-21 can inhibit the proliferation and transformation of cardiac fibroblasts induced by TGF-beta 1. Part 2 Effect of fibroblast growth factor-21 on collagen synthesis induced by TGF-beta 1 in cardiac fibroblasts Objective To investigate the effect of fibroblast growth factor-21 on transforming growth factor-beta 1. Methods The experiment was divided into blank control group, TGF-beta 1 group, TGF-beta 1 + FGF21 group. After different stimulating factors interfered with cardiac fibroblasts to a predetermined time, real-time fluorescence quantitative PCR was used to detect the mRNA of myocardial fibrosis markers CTGF, Collagen I and Collagen III. Results TGF-beta 1 significantly increased the mRNA content of myocardial fibrosis markers CTGF, Collagen I and Collagen III, and the protein expression levels of Collagen I and Collagen III were also significantly increased. ConclusionFibroblast growth factor-21 can inhibit collagen synthesis in cardiac fibroblasts induced by TGF-beta. Part III Fibroblast growth factor-21 can inhibit cardiac fibroblast collagen synthesis in patients with rheumatic heart disease and atrial fibrillation. Objective To detect the expression of fibroblast growth factor-21 in serum and atrial myocardium of patients with rheumatic heart disease and atrial fibrillation, and to explore the relationship between fibroblast growth factor-21 and atrial fibrosis in patients with rheumatic heart disease and atrial fibrillation. HE staining, Masson staining and mRNA content of myocardial fibrosis markers CTGF, Collagen I and Collagen III were used to evaluate atrial fibrosis. In addition, serum levels of FGF21, immunohistochemical staining and immunohistochemical staining were detected by ELISA kit. The expression of FGF21 in atrial myocardium was detected by real-time fluorescence quantitative PCR. Finally, the relationship between the expression of FGF21 and the severity of atrial fibrosis in patients with rheumatic heart disease was evaluated by linear correlation analysis. The mRNA levels of CTGF, Collagen I and Collagen III were significantly increased. In addition, the levels of serum FGF21 were significantly increased in patients with atrial fibrillation. Immunohistochemical staining showed that the distribution of cardiac fibroblast growth factor 21 was significantly increased in patients with atrial fibrillation, and the mRNA content of fibroblast growth factor 21 was also significantly increased. Conclusion The expression of fibroblast growth factor 21 in serum and myocardium of patients with rheumatic heart disease and atrial fibrillation is significantly increased, which is closely related to the severity of atrial fibrosis and is involved in the occurrence and maintenance of atrial fibrillation. One of.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R541.75
本文编号：2189646
[Abstract]:Part I Effects of FGF21 on cardiac fibroblasts proliferation induced by TGF-beta 1 Objective To investigate the effect of fibroblast growth factor-21 on cardiac fibroblasts proliferation and transformation induced by transforming growth factor-beta 1 (TGF-beta 1). CCK8 kit was used to detect the proliferation of cardiac fibroblasts. The expression of myofibroblast marker alpha-SMA was detected by immunofluorescence staining. The expression of PCNA and Ki67 mRNA was detected by real-time quantitative PCR. Compared with the control group, TGF-beta 1 significantly increased the proliferation rate of cardiac fibroblasts, the mRNA content of cell proliferation markers PCNA and Ki67 was significantly increased, and TGF-beta 1 significantly increased the expression of cardiac fibroblasts marker alpha-SMA. Conclusion Fibroblast growth factor-21 can inhibit the proliferation and transformation of cardiac fibroblasts induced by TGF-beta 1. Part 2 Effect of fibroblast growth factor-21 on collagen synthesis induced by TGF-beta 1 in cardiac fibroblasts Objective To investigate the effect of fibroblast growth factor-21 on transforming growth factor-beta 1. Methods The experiment was divided into blank control group, TGF-beta 1 group, TGF-beta 1 + FGF21 group. After different stimulating factors interfered with cardiac fibroblasts to a predetermined time, real-time fluorescence quantitative PCR was used to detect the mRNA of myocardial fibrosis markers CTGF, Collagen I and Collagen III. Results TGF-beta 1 significantly increased the mRNA content of myocardial fibrosis markers CTGF, Collagen I and Collagen III, and the protein expression levels of Collagen I and Collagen III were also significantly increased. ConclusionFibroblast growth factor-21 can inhibit collagen synthesis in cardiac fibroblasts induced by TGF-beta. Part III Fibroblast growth factor-21 can inhibit cardiac fibroblast collagen synthesis in patients with rheumatic heart disease and atrial fibrillation. Objective To detect the expression of fibroblast growth factor-21 in serum and atrial myocardium of patients with rheumatic heart disease and atrial fibrillation, and to explore the relationship between fibroblast growth factor-21 and atrial fibrosis in patients with rheumatic heart disease and atrial fibrillation. HE staining, Masson staining and mRNA content of myocardial fibrosis markers CTGF, Collagen I and Collagen III were used to evaluate atrial fibrosis. In addition, serum levels of FGF21, immunohistochemical staining and immunohistochemical staining were detected by ELISA kit. The expression of FGF21 in atrial myocardium was detected by real-time fluorescence quantitative PCR. Finally, the relationship between the expression of FGF21 and the severity of atrial fibrosis in patients with rheumatic heart disease was evaluated by linear correlation analysis. The mRNA levels of CTGF, Collagen I and Collagen III were significantly increased. In addition, the levels of serum FGF21 were significantly increased in patients with atrial fibrillation. Immunohistochemical staining showed that the distribution of cardiac fibroblast growth factor 21 was significantly increased in patients with atrial fibrillation, and the mRNA content of fibroblast growth factor 21 was also significantly increased. Conclusion The expression of fibroblast growth factor 21 in serum and myocardium of patients with rheumatic heart disease and atrial fibrillation is significantly increased, which is closely related to the severity of atrial fibrosis and is involved in the occurrence and maintenance of atrial fibrillation. One of.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R541.75
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