發(fā)布時間：2018-08-18 13:44

【摘要】：第一部分FGF21對TGF-β1誘導的心臟成纖維細胞增殖的影響目的探討成纖維細胞生長因子FGF21對轉化生長因子β1 (TGF-β1)誘導的心臟成纖維細胞增殖轉化的影響。方法本部分實驗分為空白對照組、TGF-β1組、FGF21組、TGF-β1+FGF21組,不同刺激因素干預心臟成纖維細胞至預定時間后,CCK8試劑盒檢測每組心臟成纖維細胞的增殖情況,細胞免疫熒光染色方法觀察每組心臟肌成纖維細胞標志物α-SMA的表達量,實時熒光定量PCR技術檢測細胞增殖標志物PCNA、Ki67的mRNA含量。結果與空白對照組相比,TGF-β1可顯著增加心臟成纖維細胞的增殖速率,細胞增殖標志物PCNA, Ki67的mRNA含量顯著提高；且TGF-β1可顯著增加心臟肌成纖維細胞標志物α-SMA的表達量。而FGF21可顯著抑制TGF-β1誘導的心臟成纖維細胞增殖、以及抑制心臟成纖維細胞向心臟肌成纖維細胞的轉化。結論成纖維細胞生長因子FGF21可抑制TGF-β1誘導的心臟成纖維細胞增殖轉化。第二部分FGF21對TGF-β1誘導的心臟成纖維細胞膠原蛋白合成的影響目的探討成纖維細胞生長因子FGF21對轉化生長因子-β1 (TGF-β1)誘導的心臟成纖維細胞膠原蛋白合成的影響。方法本部分實驗分為空白對照組、TGF-β1組、TGF-β1+FGF21組,不同刺激因素干預心臟成纖維細胞至預定時間后,實時熒光定量PCR技術檢測心肌纖維化標志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量,此外,蛋白免疫印跡法檢測Collagen Ⅰ和Collagen Ⅲ的蛋白表達水平。結果與空白對照組相比,TGF-β1可顯著增加心肌纖維化標志物CTGF、CollagenI和Collagen Ⅲ的mRNA含量；且Collagen Ⅰ和Collagen Ⅲ的蛋白表達水平也顯著提高。而FGF21可顯著抑制心肌纖維化標志物CTGF、Collagen Ⅰ和CollagenⅢ的mRNA含量、以及抑制Collagen Ⅰ和Collagen Ⅲ的蛋白表達水平。結論成纖維細胞生長因子FGF21可抑制TGF-β.誘導的心臟成纖維細胞膠原蛋白合成。第三部分FGF21在風濕性心臟病心房顫動患者心肌組織中的表達情況目的檢測風濕性心臟病心房顫動患者血清和心房肌組織中成纖維細胞生長因子FGF21的蛋白水平,探討FGF21與風濕性心臟病心房顫動患者心房纖維化的關系。方法在換瓣術中收集風濕性心臟病患者的血清和右心房肌組織,分為竇性心律組和心房顫動組。HE病理切片染色觀察每組心肌細胞橫截面積,Masson染色及心肌纖維化標志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量評估心房纖維化；此外,ELISA試劑盒檢測每組患者血清的FGF21水平,免疫組織化學染色及實時熒光定量PCR技術檢測心房肌組織中FGF21的表達水平。最后,線性相關分析法評估風濕性心臟病患者FGF21表達水平與心房纖維化嚴重程度的聯(lián)系。結果與竇性心律組相比較,心房顫動組患者的心肌細胞橫截面積、膠原容積分數、心肌纖維化標志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量均顯著增加。此外,心房顫動患者血清FGF21水平顯著升高,免疫組織化學染色顯示心房顫動患者心肌組織FGF21分布量顯著增高、且FGF21的mRNA含量也顯著升高。線性相關分析發(fā)現FGF21的表達水平與心房纖維化程度呈正相關。結論風濕性心臟病心房顫動患者血清及心肌組織中FGF21的表達顯著增加,與心房纖維化嚴重程度密切相關,參與心房顫動的發(fā)生與維持,FGF21或許可成為風濕性心臟病心房顫動患者心房纖維化的生物標志物之一。
[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.
【學位授予單位】：武漢大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R541.75

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