| Part I:The changes and clinical significance of serum level of syndecan-4protein in patients with chronic congestive heart failureBackgroundThe pathophysiology of chronic congestive heart failure(CHF) is complex syndrome,which is the end stage of development in various cardiovascular disease. CHF appears to result form myocardial damage or injury in a process known as myocardial structure and function change,which is reduced left ventricular ejection function. CHF results from myocardial degradation and pathologic structural regeneration in a process known as cardiac remodeling. This multistage and multicellular repair process is mediated by a complex interplay between growth factors and cytokines. Progressive and end-stage heart failure is a culmination of this pathologic process. Lack of objective diagnostic standards is one of the reasons of high mortality for heart failure. The recent studies show that a variety of biomarkers detection in patients with CHF has important clinical value for evaluation the diagnosis and therapeutic effect in CHF patients. The significance of NT-proBNP in heart failure has been universally recognized for international guidelines and experts from all over the world. As an important biomarker to measure the severity of chronic heart failure, serum level of NT-proBNP has been widely used in clinic. As a member of galectin family,galetin-3is a powerful pro-inflammatory factor,which can promote the infiltration of heart macrophages and stimulate the activation of fibroblasts.The macrophages and fibroblasts plays an important role in the in the process of myocardial fibrosis. The recent research show that, as a new biomarker of heart fuilure, galetin-3is a supplement to the natriuretic peptide. Syndecan-4is a member belonging to the syndecans family which are type I transmembrane proteins having a core protein modified with glycosaminoglycan chains,most commonly heparin sulphate. Syndecan-4is recognized as a "co-receptor" in conjunction with other receptors, such as growth factor, regulating the behavior of cells. It plays an important part in the cell expansion, recognition, adhesion, migration and proliferative regulation, and mediates inflammatory reaction. Syndecan-4is a transmembrane heparan sulfate-carrying glycoprotein that mediates signal transduction pathways activated by growth factors and cell surface receptors,thereby modulating tissue regeneration, angiogenesis, and focal adhesion.The recently study discovered that syndecan-4protein plays an important role in inflammatory and myocardial remodeling,both of which have been linked to chronic congestive heart failure(CHF). As a kind of inflammatory cytokines,syndecan-4protein can regulate the endothelial cell damage repair and smooth muscle cell proliferation and migration, involved in the regeneration of the vascular injury repair. It is also confirmed that the expression of syndecan-4protein is upregulated in atherosclorosis(AS) and myocardial infarction(MI) respectively,but it is unclear for the role of syndecan-4protein in patitents with chronic congestive heart failure.ObjectiveThe aim of the part I study was to investigate the significance of serum level of syndecan-4protein in patitents with chronic congestive heart failure(CHF). Furthermore,the study were also to determine whether serum level of syndecan-4protein concentration links with NT-proBNP and galetin-3concentration and to determine which echocardiographic parameters such as left ventricle ejection fraction(LVEF), fraction shortening(FS), left ventricle end-diastolic diameter(LVDd), left ventricle end-systolic diameter(LVDs), left ventricle end-diastolic volume (LVEDV) and left ventricle end-systolic volume(LVESV) are represented by serum syndecan-4protein concentration.MethodsThe concentration of serum syndecan-4protein were measured by enzyme-linked immunosorbent assay(ELISA) in40patients with CHF(62.8±11.6years,60%male) and40healthy (61.0±7.8years,57.5%male)controls(NCD) respectively. According to the NYHA class, the patients with NYHA class II, NYHA class Ⅲ, NYHA class IV are17cases,11cases and13cases, respectively. The present study excluded patients with a history of neoplastic, hepatic, infectious, collagen, or peripheral vascular atherosclerotic diseases or patients who underwent any surgical procedure in the recently6months. The parameters of left ventricle ejection fraction(LVEF), fraction shortening(FS), left ventricle end-diastolic diameter(LVDd), left ventricle end-systolic diameter(LVDs), left ventricle end-diastolic volume (LVEDV) and left ventricle end-systolic volume(LVESV) were detected by echocardiography in40patients of CHF,respectively. Results The levels of serum syndecan-4protein significantly increased in CHF group as compared with control group (P<0.01). In CHF group, as the increased grading of cardiac function according to NewYork heart association class (NYHA), the levels of serum syndecan-4protein significantly increased (P<0.05). The more upgraded the NYHA grading, the higher level of serum syndecan-4protein. In bivariate correlations analysis showed that the level of serum syndecan-4protein was significantly negative correlated with LVEF and FS(P<0.05) and was significantly positive correlated with LVDd, LVDs, LVEDV and LVESV,respectively(P<0.05).ConclusionSince serum syndecan-4protein concentration was significantly increased in patients with chronic congestive heart failure, the level of serum syndecan-4protein may have an important value in detection and surveillance of chronic congestive heart failure. Part II:Effect of tumor necrosis factor-a on cell proliferation and expression of syndecan-4protein in neonatal rat cardiac fibroblasts in vitroBackground Cardiac fibroblasts(CFs) are the most abundant cell type in the human heart, which account for60～70%of the cells. They produce matrix components and other mediators which modulate myocardial remodeling. CFs are a key source of components of the extracellular matrix (ECM) that regulates the structure of the heart and hence mechanical, chemical and electrical signals between the cellular and non-cellular components. Many of the functional effects of cardiac fibroblasts are mediated through differentiation to a myofibroblast phenotype that expresses contractile proteins and exhibits increased migratory, proliferative and secretory properties. Cardiac myofibroblasts respond to proinfiammatory cytokines (eg. TNF-a, IL-1, IL-6, TGF-β), vasoactive peptides (eg. angiotensin II, endothelin-1, natriuretic peptides) and hormones (eg. noradrenaline), the levels of which are increased in the remodeling heart. Myocardial remodeling including myocardial cells and extracellular matrix remodeling is one of the major physiopathologic mechanism in CHF. Since myocardial fibrosis is the most important factor in the extracellular matrix remodeling, reversing myocardial fibrosis has become a hotspot in current researches. Syndecan-4is recognized as a "co-receptor" in conjunction with other receptors, such as growth factor, regulating the behavior of cells. It plays an important part in the cell expansion, recognition, adhesion, migration and proliferative regulation, and mediates inflammatory reaction. Syndecan-4is a transmembrane heparan sulfate-carrying glycoprotein that mediates signal transduction pathways activated by growth factors and cell surface receptors,thereby modulating tissue regeneration, angiogenesis, and focal adhesion. The recently study discovered that syndecan-4plays an important role in inflammatory and myocardial remodeling, both of which have been linked to chronic congestive heart failure(CHF). TNF-a is produced by mononuclear macrophages and associated with myocardial fibrosis. TNF-a is a proinflammatory cytokine that is upregulated in many cardiac diseases such as ischemic human hearts, myocardial infarction and heart failure. The elevation of TNF-α is associated with cardiac hypertrophy and dysfunction, and reduced survival. The increase of TNF-α expression affects both heart function and the structure of the extracellular matrix. TNF-α plays an important role in the cardiac remodeling of CHF, but it is unclear for the effect of TNF-α on cell proliferation and expression of syndecan-4protein in neonatal rat cardiac fibroblasts in vitro.ObjectiveTumor necrosis factor-alpha (TNF-α) is one of critical regulatory factors in inflammatory reaction that plays an important role in the development of CHF. The aim of the part Ⅱ study was to investigate the effect of TNF-α on cell proliferation and expression of syndecan-4protein in neonatal rat cardiac fibroblasts in vitro.Methods1. The detection of proliferation of neonatal rat cardiac fibroblastsCardiac fibroblasts of neonatal Sprague-Daw-ley rats were cultured in96well assay in vitro and stimulated by5ng/mL TNF-α,10ng/mL TNF-α,20ng/mL TNF-α,30ng/mL TNF-α respectively. All the groups were cultured for24hours, in addition to the untreated control group established for comparison. We repeated the experiment3times under the same conditions, then we got150data in total. The non-radioactive MTS/PMS assay was adopted to detect the ratio of proliferation in neonatal Sprague-Daw-ley rats CFs.2. The detection of expression of syndecan-4protein in neonatal rat cardiac fibroblastsCardiac fibroblasts of neonatal Sprague-Daw-ley rats were cultured in vitro and stimulated by5ng/mL TNF-α,10ng/mL TNF-α,20ng/mL TNF-α,30ng/mL TNF-α respectively, the untreated control group established for comparison. The cells were lysed and the concentration of protein was evaluated by BCA Kit. The expression of syndecan-4protein in neonatal rat CFs was detected by Western blot using syndecan-4antibody. The experiment was repeated3times and we got15data in total. The expression of syndecan-4protein was represented by the intensity.ResultsSyndecan-4is expressed in the nucleus and cytoplasm of cardiac fibroblasts. Compared to the control group, TNF-a had no effect on the proliferation of neonatal rat cardiac fibroblasts at the concentration of5ng/mL and lOng/mL respectively(P>0.05), but it can significantly stimulated the proliferation of neonatal rat cardiac fibroblasts at the concentration of20ng/mL and30ng/mL respectively (P <0.05). Compared to the control group, the expression of syndecan-4protein in neonatal rat cardiac fibroblasts was not enhanced by TNF-a at the concentration of5ng/mL and10ng/mL respectively (P>0.05), but TNF-a can significantly stimulated the expression of syndecan-4protein in neonatal rat cardiac fibroblasts at the concentration of20ng/mL and30ng/mL respectively (P<0.05).ConclusionSyndecan-4is expressed in the nucleus and cytoplasm of cardiac fibroblasts. TNF-a can stimulate the cell proliferation and the expression of syndecan-4protein in of neonatal cardiac fibroblasts in vitro. |
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