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Silencing of c-Ski augments TGF-beta 1-induced epithelial-mesenchymal transition in cardiomyocyte H9C2 cells

期刊: CARDIOLOGY JOURNAL, 2019; 26 (1)

Background: The shRNA lentiviral vector was constructed to silence c-Ski expression in cardiac muscle cells, with the aim of exploring the role of c-Ski in transforming growth factor beta 1 (TGF-beta 1)-induced epithelial-mesenchymal transitions (EMT) in H9C2 cells. Methods: Real-time polymerase chain reaction (RT-PCR) and western blot were used to detect c-Ski expression at protein and messenger ribonucleic acid (mRNA) levels in 5 different cell lines. Then, lentiviral vector was constructed to silence or overexpress c-Ski in H9C2 cells. MTT and/or soft agar assay and transwell assay were used to detect cell proliferation and migration, respectively. The expression levels of c-Ski under different concentrations of TGF-beta 1 stimulation were detected by RT-qPCR and immunocytochemical analysis. In the presence or absence of TGF-beta 1 stimulation, the proteins' expression levels of alpha-SMA, FN and E-cadherin, which are closely correlated with the process of EMT, were measured by western blot after c-Ski silencing or overexpression. Meanwhile, the effect of c-Ski on Samd3 phosphorylation with TGF-b1 stimulation was investigated. Results: There is a high expression of c-Ski at protein and mRNA levels in H9C2 cell line, which first demonstrated the presence of c-Ski expression in H9C2 cells. Overexpression of c-Ski significantly increased H9C2 cell proliferation. The ability of c-Ski gene silencing to suppress cell proliferation was gradually enhanced, and inhibition efficiency was the highest after 6 to 7 d of transfection. Moreover, H9C2 cells with c-Ski knockdown gained significantly aggressive invasive potential when compared with the control group. TGF-beta 1 stimulation could dose-independently reduce c-Ski expression in H9C2 cells and lead to obvious down-regulated expression of E-cadherin. Interestingly, c-Ski could restore E-cadherin expression while suppressing alpha-SMA and/or FN expression stimulated by TGF-beta 1. However, shRNA-induced c-Ski knockdown aggravated only the TGF-beta 1-induced EMT. Moreover, c-Ski-shRNA also promoted the phosphorylation of Samd3 induced by TGF- beta 1. Conclusions: c-Ski expression in cardiac muscle cells could be down-regulated by TGF- beta 1. Silencing of c-Ski gene was accompanied by down-regulation of E-cadherin, up-regulation of alpha-SMA and/or FN and Smad3 phosphorylation induced by TGF- beta 1, promoting EMT process. Therefore, c-Ski may be closely associated with TGF-beta 1-induced EMT and play an important role in cardiac fibrosis development and progression.

JIF:1.74

Loss of AMIGO2 causes dramatic damage to cardiac preservation after ischemic injury

期刊: CARDIOLOGY JOURNAL, 2019; 26 (4)

Background: Recent studies have identified amphoterin-induced gene and open reading frame (AMIGO2). The role of AMIGO2 in tumour research is well-studied, but its role in ischemic heart diseases is seldom reported. In the present study, the role of AMIGO2 in myocardial infarction (MI) is under investigation for the first time. Methods: For in vitro studies, cardiomyocytes (CMs) and endothelial cells (ECs) were isolated from both AMIGO2 knockout (KO) and WT mice. The apoptosis of CMs was tested after 48 h of ischemic stimulation. A proliferation test was implemented after 7 days of normoxic incubation and tube formation on ECs. For in vivo studies, the MI model was built in mice hearts. Echocardiographic evaluation was performed at 3 days and 28 days post-MI, while the hemodynamics test was performed at 28 days post-MI. The histological results of the apoptosis, proliferation, angiogenesis and infarct zone assessments were determined using terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) assay, Ki67 staining, alpha-SMA/CD31 immunostain and the Masson-Trichrome method, respectively. The expression changes of the Akt pathway and related proteins were confirmed using both quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Results: The present results demonstrated that AMIGO2 deficiency caused more CMs suffering apoptosis, lower proliferation and less angiogenesis in vitro and in vivo. Weaker cardiac function and larger scar formation were detected in AMIGO2 KO mice, and increased expression of active-caspase-3 and decreased expression of PDK1, p-Akt, Bcl-2/Baxand VEGF occurred. Conclusions: Herein the findings indicate that AMIGO2 deficiency plays an attenuated cardio-protective role in ischemic heart disease via inactivation of the PDK1/Pten/Akt pathway.

JIF:1.74

Predicting functional mitral stenosis after restrictive annuloplasty for ischemic mitral regurgitation

期刊: CARDIOLOGY JOURNAL, 2019; 26 (4)

Background: Although it has been realized that restrictive mitral valve annuloplasty (MVA) may result in clinically significant functional mitral stenosis (MS), it still cannot be predicted. The purpose of this study was to identify risk factors for clinically significant functional MS following restrictive MVA surgery for chronic ischemic mitral regurgitation (CIMR). Methods: One hundred and fourteen patients who underwent restrictive MVA with coronary artery bypass grafting (CABG) for treatment of CIMR were retrospectively reviewed. Clinically significant functional MS was defined as resting transmitral peak pressure gradient (PPG) >= 13 mmHg. Results: During the follow-up period (range 6-12 months), 28 (24.56%) patients developed clinically significant functional MS. The PPG at follow-up was significantly higher than that measured in the early postoperative stage (3-5 days after surgery). Moreover, there was a linear correlation between the two measurements (r = 0.398, p < 0.001). Annuloplasty size <= 27 mm and early postoperative PPG >= 7.4 mmHg could predict clinically significant functional MS at 6-12 months postoperatively. Conclusions: Chronic ischemic mitral regurgitation patients treated with restrictive MVA and CABG have significant increases in PPG postoperatively. Annuloplasty size <= 27 mm and early postoperative PPG >= 7.4 mmHg can predict clinically significant functional MS at 6-12 months after surgery.

JIF:1.74

Invasive reperfusion after 12 hours of the symptom onset remains beneficial in patients with ST-segment elevation myocardial infarction: Evidence from a meta-analysis of published data

期刊: CARDIOLOGY JOURNAL, 2019; 26 (4)

Background: Early myocardial reperfusion therapy (< 12 h) in patients with acute myocardial infarction (AMI) can significantly improve their prognosis. However, the effect of late reperfusion (> 12 h) remains controversial. In this study, the effects of late reperfusion versus standard drug therapy on the outcomes of patients with AMI were evaluated by systematic review and meta-analysis. Methods: PubMed, Embase, Medline, Cochrane, Wanfang, and CNKI databases were searched for eligible studies for the present study. Meta-analysis was performed using RevMan 5.3.3 software. Relative risk (RR) and the 95% confidence interval (CI) were used to compare the outcomes between the two groups. The main outcome measures were major adverse cardiac events (MACEs), all-cause mortality, recurrent myocardial infarction (MI), and heart failure. Results: Eighteen studies were identified including 14,677 patients, of whom 5157 received late reperfusion with percutaneous coronary intervention (PCI) and 9520 received medication therapy (MT). Compared to MT, late PCI was associated with decreased all-cause mortality (RR 0.60, 95% CI 0.44-0.83; p = 0.002), MACEs (RR 0.67; 95% CI 0.50-0.89; p < 0.001), and heart failure (RR 0.76; 95% CI 0.60-0.97; p = 0.03), while there was also a trend toward decreased recurrent MI (RR 0.70; 95% CI 0.47-1.05; p = 0.08). However, subgroup analysis according to time to PCI showed that the clinical benefit was only from PCI after 12 h but not from 2 to 60 days of the onset of symptoms. Conclusions: The present meta-analysis suggested that PCI performed > 12 h but not 2-60 days after AMI is associated with significant improvement in clinical outcomes. However, these results need further rigorously designed large sample size clinical trials to be validated.

JIF:1.74

The effect of beta-blockers on mortality in patients with heart failure and atrial fibrillation: A meta-analysis of observational cohort and randomized controlled studies

期刊: CARDIOLOGY JOURNAL, 2019; 26 (6)

Background: Beta-blockers (BB) are the cornerstone of therapy for heart failure (HF); however, the effects of these drugs on the prognosis of patients with concomitant atrial fibrillation (AF) remain controversial. The objective of this meta-analysis was to evaluate the efficacy of BB on mortality in HF coexisting with AF. Methods: A systematic search of PubMed, Embase and the Cochrane Library databases was conducted. Observational cohort studies and randomized controlled trials reporting outcomes of mortality or HF hospitalizations for patients with HF and AF, being assigned to BB treatment. A non-BB group was also included. Results: A total of 8 clinical studies (5 randomized controlled trials and 3 observational cohort studies) involving 34197 patients were included in the analysis. The pooled analysis demonstrated that BB treatment was associated with a 22% reduction in relative risk of all-cause mortality in patients with HF and AF (RR: 0.78; 95% CI 0.71-0.86; p < 0.00001; I-2 = 27%). The pooled analysis of 5 studies reported the outcome of HF hospitalization (2774 patients) which showed that BB therapy was not associated with a reduction of HF hospitalizations (RR: 0.94; 95% CI 0.79-1.11; p = 0.46; I-2 = 38%). Conclusions: Meta-analysis suggests the potential mortality benefit of BB in patients with HF and AF. It was concluded herein that it is premature to deny patients with AF and HF to receive BB therapy considering current evidence.

JIF:1.74

Right atrial pathology in arrhythmogenic right ventricular dysplasia

期刊: CARDIOLOGY JOURNAL, 2019; 26 (6)

Background: Atrial fibrillation (AF) is the most common atrial arrhythmia in arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVD). Considering the histologic changes known in the right ventricular (RV) in ARVD, the aim of the present study was to examine right atrial (RA) pathology in patients with ARVD. Methods: Histology of RA and RV was assessed from autopsy material in 3 patients with ARVD without persistent atrial arrhythmia. RA histology in 3 patients with permanent AF without ARVD and 5 patients without cardiovascular disease was also studied. Staining with hematoxylin phloxine saffron was performed for the ARVD patients to identify fibrosis, and hematoxylin-eosin for identification of lymphocytes. Masson's trichrome staining was performed for control groups taken from a collection of standard glass slides. Results: In all 3 ARVD cases, RA anomalies were observed that revealed a reduction of cardiomyocytes, the presence of adipocytes, some of them inside the mediomural atrial layer and interstitial fibrosis. In 2 ARVD cases, interstitial fibrosis was also associated with a focus of replacement fibrosis, which was also observed in patients with permanent AF without ARVD. The histologic specimen of the RA and RV from the control group without cardiovascular disease did not display any evidence of fat or fibrosis with a preserved cardiomyocyte architecture. Conclusions: A similar histopathological substrate, as can be observed in the RV of patients with ARVD can also be seen in the RA of these patients. This may explain the high prevalence of atrial arrhythmias, particularly Al; in patients with ARVD.

JIF:1.74

The impact of renal function on the prognostic value of N-terminal pro-B-type natriuretic peptide in patients with coronary artery disease

期刊: CARDIOLOGY JOURNAL, 2019; 26 (6)

Background: The impact of renal function on the prognostic value of N-terminal pro-B-type natriuretic peptide (NT-proBNP) remains unclear in coronary artery disease (CAD). This study sought to investigate the value of using NT-proBNP level to predict prognoses of CAD patients with different estimated glomerular filtration rates (eGFRs). Methods: A retrospective analysis was conducted from a single registered database. 2087 consecutive patients with CAD confirmed by coronary angiography were enrolled. The primary endpoint was all-cause mortality. Results: The mean follow-up time was 26.4 +/- 11.9 months and death events occurred in 197 cases. The NT-proBNP levels increased with the deterioration of renal function, as well as the optimal cutoff values based on eGFR stratification to predict endpoint outcome (179.4 pg/mL, 1443.0 pg/mL, 3478.0 pg/mL, for eGFR >= 90, 60-90 and < 60 mL/min/1.73 m(2), respectively). Compared with the routine cut-off value or overall optimal one, stratified optimal ones had superior predictive ability for endpoint in each eGFR group (all with the highest Youden's J statistics). And the prognostic value became weaker as eGFR level decreased (eGFR >= 90 vs. 60-90 vs. < 60 mL/min/1.73 m(2), odds ratio [OR] 7.7; 95% confidence interval [CI] 1.7-33.9 vs. OR 4.8; 95% CI 2.7-8.5 vs. OR 3.0; 95% CI 1.5-6.2). Conclusions: This study demonstrated that NT-proBNP exhibits different predictive values for prognosis for CAD patients with different levels of renal function. Among the assessed values, the NT-proBNP cut-off value determined using renal function improve the accuracy of the prognosis prediction of Moreover, lower eGFR is associated with a higher NT-proBNP cut-off value for prognostic prediction.

JIF:1.74

Prevention of in-stent restenosis with endothelial progenitor cell (EPC) capture stent placement combined with regional EPC transplantation: An atherosclerotic rabbit model

期刊: CARDIOLOGY JOURNAL, 2019; 26 (3)

Background: Even with drug-eluting stents, the risk of in-stent restenosis (ISR) remains high. The goal of this study was to investigate the use of an endothelial progenitor cell (EPC) capture stent plus regional EPC transplantation to reduce the ISR rate. Methods: Endothelial progenitor cell capture stents were fabricated using fibrin gel and anti-CD34 plus anti-VEGFR-2 dual antibodies. Twenty male New Zealand white rabbits established as an atherosclerotic model were randomly divided into two groups: group 1 (n = 10), in which EPC capture stents were deployed into the right iliac artery; and group 2 (n = 10), in which sirolimus-eluting stents were placed. In both groups, EPCs were transplanted into target vessels beyond the stents, with outflow blocked. Radiologic-pathologic correlation outcomes were reviewed after 2 months. Results: The technical success rate of EPC capture stent placement plus EPC transplantation was 100%. The ISR rate in group 1 was lower than in group 2 (1/10 vs. 4/10; p > 0.05). Minimal luminal diameters were larger in group 1 than in group 2 (computed tomographic angiography, 1.85 +/- 0.15 mm vs. 1.50 +/- 0.20 mm; duplex ultrasound, 1.90 +/- 0.10 mm vs. 1.70 +/- 0.30 mm; p > 0.05). Transplanted EPCs were tracked positively only in group 1. Pathologic analysis demonstrated neointimal hyperplasia thickness of 0.21 +/- 0.09 mm in group 1 vs. 0.11 +/- 0.07 min in group 2 (p < 0.05). Conclusion: Endothelial progenitor cell capture stent placement plus local EPC transplant decreases the ISR rate through thrombosis reduction rather than through neointimal hyperplasia inhibition.

JIF:1.74

Hyperuricemia and severity of coronary artery disease: An observational study in adults 35 years of age and younger with acute coronary syndrome

期刊: CARDIOLOGY JOURNAL, 2019; 26 (3)

Background: Coronary artery disease (CAD) in adults <= 35 years of age is rare, but the incidence is on the rise and the risk factors for this age group are largely uncertain. Previous studies have shown that hyperurkemia (HUA) is an independent risk factor for CAD in the general population, whereas the role in adults <= 35 years of age with acute coronary syndrome (ACS) is unclear. Methods: Patients, 18-35 years of age, diagnosed with ACS for the first time at the documented institution between January 2005 and December 2015, were enrolled in the current study. The severity of CAD was assessed by the Gensini score. Patients were divided into two groups according to the definition of HUA. The relationship between HUA and CAD severity was assessed based on multi-variate analysis. Results: Seven hundred seventy-one participants fulfilling the criteria were included in this study (mean age, 31.6 years; 94.4% male). HUA, which was defined as a serum uric acid level >= 7.0 mg/dL (420 mu mol/L) in males and >= 6.0 mg/dL (357 mu mol/L) in females, accounted for 37% of the participants. Multivariate analysis identified that HUA is an independent risk factor of CAD severity, as assessed by the Gensini score, in very young adults with ACS (OR 8.28; 95% CI 1.96-14.59; p = 0.01), and the effect of HUA on CAD severity was second only to diabetes mellitus. Conclusions: Hyperuricemia was shown to be an independent risk factor for CAD severity in young adults with ACS (18-35 years of age).

JIF:1.74

Long-term outcomes of mitral valve annuloplasty versus subvalvular sparing replacement for severe ischemic mitral regurgitation

期刊: CARDIOLOGY JOURNAL, 2019; 26 (3)

Background: Although practice guidelines recommend surgery for patients with severe chronic ischemic mitral regurgitation (CIMR), they do not specify whether to repair or replace the mitral valve. The purpose of this study was to evaluate the long-term outcomes in patients with severe CIMR undergoing mitral valve annuloplasty (MVA) versus subvalvular sparing mitral valve replacement (MVR). Methods: 392 consecutive patients who underwent MVA or subvalvular sparing MVR for treatment of severe CIMR were retrospectively reviewed. Results: After adjustment for baseline differences with multivariable regression analysis at 53 months follow-up (interquartile range, 34-81 months), there was no significant difference between the two groups for risk of major adverse cardiac or cerebrovascular events (MACCE), cardiac death, or all-cause death. Propensity score matching extracted 77 pairs. During the follow-up, compared with the MVR group, both the left atrium and left ventricle end-diastolic diameter were markedly larger (p = 0.013 and p = 0.033, respectively), and the incidence of mural regurgitation recurrence was significantly higher in the MVA group (p < 0.001). No significant difference was observed between the two propensity score-matched groups in composite in-hospital outcomes, overall survival, freedom from cardiac death or MACCE, except subvalvular sparing MVR was associated with a lower incidence of hospitalization f or heart failure than MVA (p = 0.015). Conclusions: Subvalvular sparing MVR is a suitable management of patients with severe CIMR, it is more favorable to ventricular remodeling and is associated with a lower incidence of hospitalization for heart failure than MVA.

JIF:1.74

Proteomics study of serum exosomes in Kawasaki disease patients with coronary artery aneurysms

期刊: CARDIOLOGY JOURNAL, 2019; 26 (5)

Background: To study the protein profile of the serum exosomes of patients with coronary artery aneurysms (CAA) caused by Kawasaki disease (KD). Methods: Two-dimensional electrophoresis (2-DE) was used to identify proteins from the exosomes of serum obtained from children with CAA caused by KD, as well as healthy controls. Differentially expressed proteins were identified using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS) analysis. Results: Thirty two differentially expressed proteins were identified (18 up-regulated and 14 downregulated) from serum exosomes of children with CAA and were compared to healthy controls. The expression levels of 4 proteins (TN RBP4, LRG1, and APOA4) were validated using Western blotting. Classification analysis and protein-protein network analysis showed that they are associated with multiple functional groups, including host immune response, inflammation, apoptotic process, developmental process, and biological adhesion process. Conclusions: These findings establish a comprehensive proteomic profile of serum exosomes from children with CAA caused by KD, and provide additional insights into the mechanisms of CAA caused by KD.

JIF:1.74

The long noncoding RNA THRIL knockdown protects hypoxia-induced injuries of H9C2 cells through regulating miR-99a

期刊: CARDIOLOGY JOURNAL, 2019; 26 (5)

Background: Myocardial infarction (MI) is a leading cause of disease with high morbidity and mortality worldwide. Recent studies have revealed that long non-coding RNAs (lncRNAs) are involved in heart disease pathogenesis. This study aimed to investigate the effect and the molecular basis of THRIL on hypoxia-injured H9C2 cells. Methods: THRIL, miR-99a and Brahma-related gene 1 (Brg1) expressions in H9C2 cells were altered by transient transfections. The cells were subjected to hypoxia for 4 h, and then the levels of THRIL, miR-99a and Brg1 were investigated. Cell viability, migration and invasion, and apoptotic cells were respectively measured by trypan blue exclusion assay, transwell migration assay and flow cytometfry assay. Dual luctferase reporter assay was conducted to verify the interaction between miR-99a and THRIL. Furthermore, levels of apoptosis-, PI3K'AKT and mTOR pathways-related factors were measured by western blotting. Results: Hypoxia induced an increase of THRIL but a reduction of miR-99a and Brg1. THRIL inhibition significantly attenuated hypoxia-induced cell injuries, as increased cell viability, migration and invasion, and decreased cell apoptosis. THRIL negatively regulated miR-99a expression through sponging with miR-99a binding site, and miR-99a inhibition abolished the protective effects of THRIL knockdown against hypoxia-induced injury in H9C2 cells. Furthermore, rniR-99a positively regulated the expression of Brg1. Brg1 inhibition promoted hypoxia-induced cell injuries, while Brg1 overexpression alleviated hypoxia-induced cell injuries. Moreover; Brg1 overexpression activated PI3K/AKT and mTOR pathways. Conclusions: This study demonstrated that THRIL inhibition represented a protective effect against hypoxia-induced injuries in H9C2 cells by up-regulating miR-99a expression.

JIF:1.74

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