【摘要】：病毒性心肌炎（viral myocarditis，VMC）是小兒最常見心血管系統(tǒng)疾病，病情輕重不一，多數(shù)患兒預(yù)后良好，部分患兒可發(fā)展為擴(kuò)張型心肌病、致殘性心律失常。VMC的發(fā)病機(jī)制尚不明確，涉及病毒的直接損傷、免疫機(jī)制、細(xì)胞凋亡、心肌纖維化、生化機(jī)制等，免疫機(jī)制一直以來是研究的熱點(diǎn)。小凹（caveolae）是定位于細(xì)胞質(zhì)膜上的膜結(jié)構(gòu)，可從細(xì)胞質(zhì)膜上脫落下來形成囊泡，把細(xì)胞外的生物活性分子通過吞飲作用向胞內(nèi)轉(zhuǎn)移、參與多種信號(hào)轉(zhuǎn)導(dǎo)過程。小凹蛋白-3（Caveolin-3）主要表達(dá)在心臟，骨骼和平滑肌的肌細(xì)胞中。研究表明， TGF-β1及其下游蛋白smad3構(gòu)成的信號(hào)通路在心肌纖維化的發(fā)生中具有重要作用；Caveolin參與冠心病、心肌病、糖尿病大血管病變等多種疾病的發(fā)展過程。在病毒性心肌炎發(fā)生機(jī)制中，Caveolin-3的作用以及Caveolin-3是否可調(diào)控TGF-β1/smad3信號(hào)通路的表達(dá)目前尚未見報(bào)道。銀杏葉提取物具有廣泛的藥理作用，如具有改善毛細(xì)血管的通透性、擴(kuò)張冠狀動(dòng)脈、改善心肌血液循環(huán)、降低心肌細(xì)胞耗氧量等多種功能，臨床已用于心血管疾病、腎病綜合征、糖尿病、老年癡呆等心腦血管疾病的預(yù)防和治療。 本研究用CVB3感染大鼠原代心肌細(xì)胞，，同時(shí)加入銀杏葉提取物，檢測銀杏葉提取物對(duì)大鼠心肌細(xì)胞的保護(hù)作用及毒性作用，同時(shí)通過western-blot、免疫組化方法檢測大鼠心肌細(xì)胞內(nèi)Cav-3、TGF-β1、smad3蛋白表達(dá)量變化。另外，通過CVB3感染Balb/C小鼠建立病毒性心肌炎小鼠模型，通過HE染色觀察小鼠心肌病理變化，通過Masson染色觀察各組小鼠心肌纖維化程度；然后分別通過RT-PCR、western-blot、免疫組化方法檢測小鼠心肌Cav-3、TGF-β1、smad3蛋白表達(dá)量及mRNA變化。體內(nèi)、體外實(shí)驗(yàn)結(jié)果顯示，Cav-3、TGF-β1及smad3蛋白在病毒感染后表達(dá)增加，銀杏葉提取物能夠改善小鼠心肌病理變化，降低心肌纖維化程度，銀杏葉提取物是通過下調(diào)Cav-3、TGF-β1及其下游蛋白smad3的表達(dá)，以實(shí)現(xiàn)治療病毒性心肌炎的作用，其治療效果與參麥注射液相當(dāng)，為臨床應(yīng)用銀杏葉提取物治療VMC提供了理論和實(shí)驗(yàn)依據(jù)。 本研究創(chuàng)新點(diǎn)：（1）Cav-3、TGF-β1及smad3蛋白在病毒性心肌炎中的作用少見報(bào)道，本研究顯示其在病毒性心肌炎發(fā)病中具有一定的作用；（2）本研究探討了銀杏提取物對(duì)病毒性心肌炎中的Cav-3、TGF-β1和smad3蛋白表達(dá)的影響，及其對(duì)病毒性心肌炎的治療作用。
[Abstract]:Viral myocarditis (viral myocarditis,VMC) is the most common cardiovascular disease in children with different severity and good prognosis. Some children can develop dilated cardiomyopathy. The pathogenesis of disabling arrhythmia. The direct damage of virus, immune mechanism, apoptosis, myocardial fibrosis, biochemical mechanism and so on, immune mechanism has been a hot topic. Concave (caveolae) is a membrane structure located on the cytoplasmic membrane, which can be removed from the cytoplasmic membrane to form vesicles, and the extracellular bioactive molecules are transferred to the cell through the action of ingestion and participate in many signal transduction processes. Concave protein-3 (Caveolin-3) is mainly expressed in muscle cells of heart, bone and smooth muscle. It has been shown that the signaling pathway of TGF- 尾 1 and its downstream protein smad3 plays an important role in the pathogenesis of myocardial fibrosis. Caveolin is involved in the development of coronary heart disease, cardiomyopathy and diabetic macroangiopathy. The role of Caveolin-3 in the pathogenesis of viral myocarditis and whether Caveolin-3 can regulate the expression of TGF- 尾 1/smad3 signaling pathway have not been reported. Ginkgo biloba extract has a wide range of pharmacological effects, such as improving capillary permeability, dilating coronary artery, improving myocardial blood circulation, reducing myocardial oxygen consumption and so on. Prevention and treatment of cardiovascular and cerebrovascular diseases such as nephrotic syndrome, diabetes mellitus and senile dementia. The protective and toxic effects of ginkgo biloba extract (Ginkgo biloba extract) on rat cardiomyocytes were detected by CVB3 infection in primary rat cardiomyocytes at the same time. At the same time, the expression of Cav-3,TGF- 尾 1 and smad3 protein in rat cardiomyocytes was detected by western-blot, immunohistochemical method. In addition, the model of viral myocarditis was established in Balb/C mice infected with CVB3, the pathological changes of myocardium were observed by HE staining, and the degree of myocardial fibrosis in each group was observed by Masson staining. RT-PCR,western-blot, immunohistochemical method was used to detect the expression of Cav-3,TGF- 尾 1 and mRNA in myocardium of mice. In vivo and in vitro, the results showed that the expression of TGF- 尾 1 and smad3 protein increased after virus infection. Ginkgo biloba extract could improve the pathological changes of myocardium and decrease the degree of myocardial fibrosis in mice. Ginkgo biloba extract can treat viral myocarditis by down-regulating the expression of Cav-3,TGF- 尾 _ 1 and its downstream protein smad3. The therapeutic effect of ginkgo biloba extract is similar to that of Shenmai injection, which provides theoretical and experimental basis for clinical application of ginkgo biloba extract in the treatment of VMC. The innovative points of this study are as follows: (1) the role of Cav-3,TGF- 尾 1 and smad3 protein in viral myocarditis is rarely reported, and this study shows that it plays a certain role in the pathogenesis of viral myocarditis; (2) the effect of ginkgo biloba extract on the expression of Cav-3,TGF- 尾 1 and smad3 protein in viral myocarditis and its therapeutic effect on viral myocarditis were studied.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R285.5;R725.4

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 王倩倩;WISP1蛋白在病毒性心肌炎中的作用及葛根素的干預(yù)作用研究[D];吉林大學(xué);2013年

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