【摘要】：第一部分FGF21對(duì)TGF-β1誘導(dǎo)的心臟成纖維細(xì)胞增殖的影響目的探討成纖維細(xì)胞生長(zhǎng)因子FGF21對(duì)轉(zhuǎn)化生長(zhǎng)因子β1 (TGF-β1)誘導(dǎo)的心臟成纖維細(xì)胞增殖轉(zhuǎn)化的影響。方法本部分實(shí)驗(yàn)分為空白對(duì)照組、TGF-β1組、FGF21組、TGF-β1+FGF21組,不同刺激因素干預(yù)心臟成纖維細(xì)胞至預(yù)定時(shí)間后,CCK8試劑盒檢測(cè)每組心臟成纖維細(xì)胞的增殖情況,細(xì)胞免疫熒光染色方法觀察每組心臟肌成纖維細(xì)胞標(biāo)志物α-SMA的表達(dá)量,實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)細(xì)胞增殖標(biāo)志物PCNA、Ki67的mRNA含量。結(jié)果與空白對(duì)照組相比,TGF-β1可顯著增加心臟成纖維細(xì)胞的增殖速率,細(xì)胞增殖標(biāo)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(xì)胞標(biāo)志物α-SMA的表達(dá)量。而FGF21可顯著抑制TGF-β1誘導(dǎo)的心臟成纖維細(xì)胞增殖、以及抑制心臟成纖維細(xì)胞向心臟肌成纖維細(xì)胞的轉(zhuǎn)化。結(jié)論成纖維細(xì)胞生長(zhǎng)因子FGF21可抑制TGF-β1誘導(dǎo)的心臟成纖維細(xì)胞增殖轉(zhuǎn)化。第二部分FGF21對(duì)TGF-β1誘導(dǎo)的心臟成纖維細(xì)胞膠原蛋白合成的影響目的探討成纖維細(xì)胞生長(zhǎng)因子FGF21對(duì)轉(zhuǎn)化生長(zhǎng)因子-β1 (TGF-β1)誘導(dǎo)的心臟成纖維細(xì)胞膠原蛋白合成的影響。方法本部分實(shí)驗(yàn)分為空白對(duì)照組、TGF-β1組、TGF-β1+FGF21組,不同刺激因素干預(yù)心臟成纖維細(xì)胞至預(yù)定時(shí)間后,實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)心肌纖維化標(biāo)志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量,此外,蛋白免疫印跡法檢測(cè)Collagen Ⅰ和Collagen Ⅲ的蛋白表達(dá)水平。結(jié)果與空白對(duì)照組相比,TGF-β1可顯著增加心肌纖維化標(biāo)志物CTGF、CollagenI和Collagen Ⅲ的mRNA含量；且Collagen Ⅰ和Collagen Ⅲ的蛋白表達(dá)水平也顯著提高。而FGF21可顯著抑制心肌纖維化標(biāo)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(dá)水平。結(jié)論成纖維細(xì)胞生長(zhǎng)因子FGF21可抑制TGF-β.誘導(dǎo)的心臟成纖維細(xì)胞膠原蛋白合成。第三部分FGF21在風(fēng)濕性心臟病心房顫動(dòng)患者心肌組織中的表達(dá)情況目的檢測(cè)風(fēng)濕性心臟病心房顫動(dòng)患者血清和心房肌組織中成纖維細(xì)胞生長(zhǎng)因子FGF21的蛋白水平,探討FGF21與風(fēng)濕性心臟病心房顫動(dòng)患者心房纖維化的關(guān)系。方法在換瓣術(shù)中收集風(fēng)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(dòng)組。HE病理切片染色觀察每組心肌細(xì)胞橫截面積,Masson染色及心肌纖維化標(biāo)志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量評(píng)估心房纖維化；此外,ELISA試劑盒檢測(cè)每組患者血清的FGF21水平,免疫組織化學(xué)染色及實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)心房肌組織中FGF21的表達(dá)水平。最后,線性相關(guān)分析法評(píng)估風(fēng)濕性心臟病患者FGF21表達(dá)水平與心房纖維化嚴(yán)重程度的聯(lián)系。結(jié)果與竇性心律組相比較,心房顫動(dòng)組患者的心肌細(xì)胞橫截面積、膠原容積分?jǐn)?shù)、心肌纖維化標(biāo)志物CTGF、Collagen Ⅰ和Collagen Ⅲ的mRNA含量均顯著增加。此外,心房顫動(dòng)患者血清FGF21水平顯著升高,免疫組織化學(xué)染色顯示心房顫動(dòng)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(guān)分析發(fā)現(xiàn)FGF21的表達(dá)水平與心房纖維化程度呈正相關(guān)。結(jié)論風(fēng)濕性心臟病心房顫動(dòng)患者血清及心肌組織中FGF21的表達(dá)顯著增加,與心房纖維化嚴(yán)重程度密切相關(guān),參與心房顫動(dòng)的發(fā)生與維持,FGF21或許可成為風(fēng)濕性心臟病心房顫動(dòng)患者心房纖維化的生物標(biāo)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(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R541.75

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