摘要
本实验的目的是为了探讨CVB3诱导的病毒性心肌炎小鼠脂联素的表达及缬沙坦对其表达的影响。实验采用4周龄的80只ICR雄性小鼠,体重约16-18g,随机分为对照组(n=20)、病毒组(n=30)和治疗组(n=30)。对照组每只小鼠连续3天腹腔注射无菌生理盐水0.2ml,病毒组和治疗组每只小鼠腹腔接种10-6TCID50柯萨奇B3病毒液0.2ml,均连续3天。治疗组小鼠接种CVB33天后,给予10mg/kg﹒day的缬沙坦灌胃,其它两组给以无菌生理盐水0.2ml灌胃。分别在实验的第7、14、21天每组随机抽取5只小鼠经眼球采血后断颈处死,留取血清、心肌标本。实验结果如下:1.成功建立了病毒性心肌炎小鼠模型;2.病毒性心肌炎小鼠的心肌病理学改变明显,可见心肌组织坏死和大量炎性细胞浸润,部分心肌出现断裂及纤维化,接种病毒第7天后心肌病变最明显,后炎症细胞逐渐减少,至第21天仅见少量炎性细胞,但可见心肌纤维化,经缬沙坦治疗后上述病变有改善趋势;3.与其它两组比较,病毒组小鼠血清脂联素水平最低,而肿瘤坏死因子水平最高;4.与其它两组比较,治疗组中心肌脂联素含量最高,病毒与治疗组心肌脂联素与心肌病理积分均呈负相关;5.三组小鼠心肌中脂联素及PPARα均呈正相关。所得结论如下:1.VMC时小鼠血清呈低脂联素水平,可为VMC的血清学检测提供一个参考指标;2.VMC时脂联素可抑制肿瘤坏死因子的表达,减少心肌炎症细胞的浸润;3.缬沙坦可提高心肌中脂联素的表达,减轻心肌炎症病变;4.过氧化物酶体增殖物激活受体可能为脂联素的信号转导通路之一。
VMC is a direct violation of the virus in the heart of myocarditis caused by transmitted diseases into the main performance, sometimes involving endocardium or pericardium, the pathological features of myocardial cell necrosis and degeneration, and even myocardial fibrosis, cardiac remodeling, and ultimately the development of into dilated cardiomyopathy. Pathogenesis of this disease is not yet entirely clear, both at home and abroad on the pathogenesis of this disease from the molecular virology, molecular immunology, etc. were studied. Tumor necrosis factor (TNF-α)by mononuclear macrophages, and inflammation are the body's immune system play an important regulatory factors, it can directly undermine the myocardial cells, resulting in interleukin abnormal, so that the immune regulatory network dysfunction, myocardial cells and vascular endothelial cell adhesion molecule expression in the immune increase,and promote substantial lymphocyte infiltration of the myocardial cells. Adiponectin is a protein secreted by adipose tissue, the present study suggests that with a wide variety of cardiovascular diseases. Coronary heart disease, heart failure and hypertension plasma adiponectin levels were low fat, and studies have shown that the plasma was low-fat joint occurs when levels of cardiovascular disease risk. Kenei and others studies show that Wulong Tea can increase coronary heart disease in patients with plasma adiponectin levels.β-adrenergic receptor agonists, adenylate cyclase agonists and glucocorticoids can be reduced the expression of adiponectin gene ,butβ-blockers, ACEI and ARB drugs could increase the plasma adiponectin levels.Ouchi found that human aortic endothelial cells by adiponectin after treatment can inhibit TNF-α-induced monocyte adhesion and adhesion molecule expression. Ouchi and Kihara,s study found that adiponectin through the activation of cyclic adenosine monophosphate (cAMP) protein kinase A (PKA) signaling pathway can inhibition of nuclear factor-kB activation and suppress TNF-αinduced nuclear factor-kB inhibitor of cytosolic phosphorylation, and thus to control the inflammatory response of endothelial cells. Physiological concentrations of adiponectin can reduce macrophage secretion of TNF-αand IL-6, and dose-dependent manner to inhibit TNF-αinduced intracellular adhesion molecule-1, E-selectin and vascular cell adhesion molecule-1 in Human aortic endothelial cell surface expression. Recent studies have shown that adiponectin can activate AMPK signaling inhibited myocardial ischemia - reperfusion injury in cardiomyocyte apoptosis, and pass COX2-PGE2 pathway inhibitor in ischemic myocardium of the secretion of TNF-a levels and reduce the extent of viral myocarditis patients, Endothelin -1 inhibition after myocardial infarction related cardiac hypertrophy. Viral myocarditis is to develop the most serious consequences as dilated cardiomyopathy, Liu and others to the pathogenesis of VMC into viral replication period, period of its own immune response in patients with dilated cardiomyopathy and three phase. When the virus will trigger a violation of myocardial a series of immune responses, while the release of a large number of inflammatory cytokines such as IL-1, 2,6,12, tumor necrosis factor, interferon, etc., to stimulate the myocardium, myocarditis caused by disease. At this point will cause the heart energy metabolism. Inflammatory stimulation of energy metabolism at the same time together with the obstacles, lead to cardiac overload, cardiac function and the onset heart failure, the load process of a long time also led to cardiac hypertrophy, cardiac remodeling and expansion into the development of cardiomyopathy. Prompted the current study of adiponectin through its receptor may activate peroxisome proliferator-activated receptor (PPARα) and AMPK signal transduction pathway of two of its physiological role.
These two signaling pathway itself with heart inflammation and cardiac hypertrophy, myocardial fibrosis and other closely related. Adiponectin has anti-inflammatory and the impact of the dual role of myocardial remodeling, and the physical conditions and metabolic effects of myocardial development. Therefore, adiponectin further defined itself and its signal transduction pathway and the relationship between viral myocardial contribute to the treatment of viral myocarditis to provide a new target.
Selection of the experimental mice infected with CVB3 as a pathogen, the success of the established model of viral myocarditis. The establishment of the control group, virus group and treatment group, drug use and effect of valsartan on CVB3 infected mice by treatment. CVB3 infection in 7,14,21 days after admission with myocardial tissue detected by immunohistochemistry myocardial cells adiponectin and peroxisome proliferator-activated receptor content, while serum adiponectin and tumor necrosis factor -αcontent.
Histopathologic examination of myocardial line check, the calculation points of myocardial pathology, statistics of the observed relationship between volume to explore the viral myocarditis in mice the expression of adiponectin and its expression of valsartan, for research on the pathogenesis of VMC progress and treatment to provide a new theoretical basis and experimental evidence.
Results: 1. Successfully established a mouse model of viral myocarditis; 2. Viral myocarditis myocardial histopathological changes in mice clearly shows that a large number of myocardial necrosis and inflammatory cell infiltration, and some faults appear and myocardial fibrosis, virus inoculation the first 7 days after the most obvious cardiomyopathy, a gradual reduction of inflammatory cells, only 21 days to see a small number of inflammatory cells, but myocardial fibrosis can be seen by valsartan treatment improved the above-mentioned diseases trends; 3. Compared with the control and treatment group comparison, the virus of mice the level of serum adiponectin minimum, while the highest levels of TNF-α. 4. Compared with the control and treatment group, compared to treatment group, myocardial highest adiponectin, viruses, and the treatment group and myocardial adiponectin were negatively correlated with pathological points, statistically significant difference. 5. Cardiomyopathy in three groups of mice and PPAR adiponectin was positively correlated, statistically significant difference. Conclusion: 1.VMC when serum adiponectin levels were low-fat for serological detection of VMC to provide a reference.
2.VMC adiponectin inhibited the expression of TNF-αto reduce the infiltration of cells in myocarditis patients.
3. Valsartan increase in myocardial expression of adiponectin to reduce the disease myocarditis lesions.
4. PPARαmay be one of the signal transduction pathway of adiponectin .
引文
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