Caveolin-3在病毒性心肌炎小鼠中的作用及银杏叶提取物干预的研究
发布时间：2018-09-05 18:02
【摘要】：病毒性心肌炎（viral myocarditis，VMC）是小儿最常见心血管系统疾病，病情轻重不一，多数患儿预后良好，部分患儿可发展为扩张型心肌病、致残性心律失常。VMC的发病机制尚不明确，涉及病毒的直接损伤、免疫机制、细胞凋亡、心肌纤维化、生化机制等，免疫机制一直以来是研究的热点。小凹（caveolae）是定位于细胞质膜上的膜结构，可从细胞质膜上脱落下来形成囊泡，把细胞外的生物活性分子通过吞饮作用向胞内转移、参与多种信号转导过程。小凹蛋白-3（Caveolin-3）主要表达在心脏，骨骼和平滑肌的肌细胞中。研究表明， TGF-β1及其下游蛋白smad3构成的信号通路在心肌纤维化的发生中具有重要作用；Caveolin参与冠心病、心肌病、糖尿病大血管病变等多种疾病的发展过程。在病毒性心肌炎发生机制中，Caveolin-3的作用以及Caveolin-3是否可调控TGF-β1/smad3信号通路的表达目前尚未见报道。银杏叶提取物具有广泛的药理作用，如具有改善毛细血管的通透性、扩张冠状动脉、改善心肌血液循环、降低心肌细胞耗氧量等多种功能，临床已用于心血管疾病、肾病综合征、糖尿病、老年痴呆等心脑血管疾病的预防和治疗。 本研究用CVB3感染大鼠原代心肌细胞，，同时加入银杏叶提取物，检测银杏叶提取物对大鼠心肌细胞的保护作用及毒性作用，同时通过western-blot、免疫组化方法检测大鼠心肌细胞内Cav-3、TGF-β1、smad3蛋白表达量变化。另外，通过CVB3感染Balb/C小鼠建立病毒性心肌炎小鼠模型，通过HE染色观察小鼠心肌病理变化，通过Masson染色观察各组小鼠心肌纤维化程度；然后分别通过RT-PCR、western-blot、免疫组化方法检测小鼠心肌Cav-3、TGF-β1、smad3蛋白表达量及mRNA变化。体内、体外实验结果显示，Cav-3、TGF-β1及smad3蛋白在病毒感染后表达增加，银杏叶提取物能够改善小鼠心肌病理变化，降低心肌纤维化程度，银杏叶提取物是通过下调Cav-3、TGF-β1及其下游蛋白smad3的表达，以实现治疗病毒性心肌炎的作用，其治疗效果与参麦注射液相当，为临床应用银杏叶提取物治疗VMC提供了理论和实验依据。 本研究创新点：（1）Cav-3、TGF-β1及smad3蛋白在病毒性心肌炎中的作用少见报道，本研究显示其在病毒性心肌炎发病中具有一定的作用；（2）本研究探讨了银杏提取物对病毒性心肌炎中的Cav-3、TGF-β1和smad3蛋白表达的影响，及其对病毒性心肌炎的治疗作用。
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R285.5;R725.4
本文编号：2225035
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R285.5;R725.4
【引证文献】
相关硕士学位论文 前1条
1 王倩倩;WISP1蛋白在病毒性心肌炎中的作用及葛根素的干预作用研究[D];吉林大学;2013年
本文编号：2225035
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