Intravenously transplanted mesenchymal stromal cells: a new endocrine reservoir for cardioprotection.

BACKGROUND: Intravenous administration of mesenchymal stromal cells (MSCs) has an acknowledged competence of cardiac repair, despite a lack of systematic description of the underlying biological mechanisms. The lung, but not the heart, is the main trapped site for intravenously transplanted MSCs, which leaves a spatial gap between intravenously transplanted MSCs and the injured myocardium. How lung-trapped MSCs after intravenous transplantation rejuvenate the injured myocardium remains unknown. METHODS: MSCs were isolated from human placenta tissue, and DF-MSCs or Gluc-MSCs were generated by transduced with firefly luciferase (Fluc)/enhanced green fluorescence protein (eGFP) or Gaussia luciferase (Gluc) lactadherin fusion protein. The therapeutic efficiency of intravenously transplanted MSCs was investigated in a murine model of doxorubicin (Dox)-induced cardiotoxicity. Trans-organ communication from the lung to the heart with the delivery of blood was investigated by testing the release of MSC-derived extracellular vesicles (MSC-EVs), and the potential miRNA inner MSC-EVs were screened out and verified. The potential therapeutic miRNA inner MSC-EVs were then upregulated or downregulated to assess the further therapeutic efficiency RESULTS: Dox-induced cardiotoxicity, characterized by cardiac atrophy, left ventricular dysfunction, and injured myocardium, was alleviated by consecutive doses of MSCs. These cardioprotective effects might be attributed to suppressing GRP78 triggering endoplasmic reticulum (ER) stress-induced apoptosis in cardiomyocytes. Our results confirmed that miR-181a-5p from MSCs-derived EVs (MSC-EVs) inhibited GRP78. Intravenous DF-MSCs were trapped in lung vasculature, secreted a certain number of EVs into serum, which could be confirmed by the detection of eGFP+ EVs. GLuc activity was increased in serum EVs from mice administrated with GLuc-MSCs. MiR-181a-5p, inhibiting GRP78 with high efficacy, was highly expressed in serum EVs and myocardium after injecting consecutive doses of MSCs into mice treated with Dox. Finally, upregulation or downregulation of miR-181a-5p levels in MSC-EVs enhanced or weakened therapeutic effects on Dox-induced cardiotoxicity through modulating ER stress-induced apoptosis. CONCLUSIONS: This study identifies intravenously transplanted MSCs, as an endocrine reservoir, to secrete cardioprotective EVs into blood continuously and gradually to confer the trans-organ communication that relieves Dox-induced cardiotoxicity.

Endogenous CCN5 Participates in Angiotensin II/TGF-ß1 Networking of Cardiac Fibrosis in High Angiotensin II-Induced Hypertensive Heart Failure.

Aberrant activation of angiotensin II (Ang II) accelerates hypertensive heart failure (HF); this has drawn worldwide attention. The complex Ang II/transforming growth factor (TGF)-ß1 networking consists of central mechanisms underlying pro-fibrotic effects; however, this networking still remains unclear. Cellular communication network 5 (CCN5), known as secreted matricellular protein, mediates anti-fibrotic activity by inhibiting fibroblast-to-myofibroblast transition and the TGF-ß1 signaling pathway. We hypothesized that endogenous CCN5 plays an essential role in TGF-ß1/Ang II networking-induced cardiac fibrosis (CF), which accelerates the development of hypertensive HF. This study aimed to investigate the potential role of CCN5 in TGF-ß1/Ang II networking-induced CF. Our clinical retrospective study demonstrated that serum CCN5 decreased in hypertensive patients, but significantly increased in hypertensive patients taking oral angiotensin-converting enzyme inhibitor (ACEI). A negative association was observed between CCN5 and Ang II in grade 2and 3 hypertensive patients receiving ACEI treatment. We further created an experimental model of high Ang II-induced hypertensive HF. CCN5 was downregulated in the spontaneously hypertensive rats (SHRs) and increased via the inhibition of Ang II production by ACEI. This CCN5 downregulation may activate the TGF-ß1 signaling pathway, which promotes direct deposition of the extracellular matrix (ECM) and fibroblast-to-myofibroblast transition via activated Smad-3. Double immunofluorescence staining of CCN5 and cell markers of cardiac tissue cell types suggested that CCN5 was mainly expressed in the cardiac fibroblasts. Isolated cardiac fibroblasts were exposed to Ang II and transfected with small interfering RNA targeting CCN5. The expression of TGF-ß1 together with Col Ia and Col IIIa was further promoted, and alpha-smooth muscle actin (α-SMA) was strongly expressed in the cardiac fibroblasts stimulated with Ang II and siRNA. In our study, we confirmed the anti-fibrotic ability of endogenous CCN5 in high Ang II-induced hypertensive HF. Elevated Ang II levels may decrease CCN5 expression, which subsequently activates TGF-ß1 and finally promotes the direct deposition of the ECM and fibroblast-to-myofibroblast transition via Smad-3 activation. CCN5 may serve as a potential biomarker for estimating CF in hypertensive patients. A novel therapeutic target should be developed for stimulating endogenous CCN5 production.

Serum VEGF: Diagnostic Value of Acute Coronary Syndrome from Stable Angina Pectoris and Prognostic Value of Coronary Artery Disease.

BACKGROUND: Although the level of serum vascular endothelial growth factor (VEGF) is elevated in coronary artery disease (CAD) patients, its potential role in acute coronary syndrome (ACS) or stable angina pectoris (SAP) patients remains unclear. OBJECTIVES: To evaluate diagnostic accuracy of serum VEGF in determining ACS patients from SAP and analyze the association of serum VEGF with coronary artery lesions in SAP or the GRACE score in ACS, which is involved in the poor prognosis of low serum VEGF. METHODS: 248 CAD patients and 48 healthy subjects were enrolled in this study. Serum VEGF levels were detected by using ELISA. The Gensini score or GRACE score was calculated among SAP or ACS patients. All the patients were followed up for a period of 12 months (mean: 10.77 months). RESULTS: VEGF serum concentrations were higher in the ACS subgroup than in the SAP subgroup (P < 0.001) with diagnostic accuracy of ACS from SAP (AUC: 0.667, sensitivity: 68.5%, specificity: 60.1%, P < 0.001). Patients with high risk of Gensini score showed reduced VEGF levels (P < 0.001) accompanied by a negative correlation (r = -0.396, P < 0.001). Patients with a higher GRACE score indicated lower VEGF levels (P < 0.001). Low serum VEGF was one of the potential risk factors with adjusted HR of 0.531 (P=0.048). CONCLUSION: Serum VEGF exhibits efficient diagnostic value for detection of ACS from SAP with a cutoff value of 648.75 pg/mL. Low serum VEGF indicates severe coronary artery lesions and a higher GRACE score, which suggests poor clinical outcomes.

Perindopril Improves Cardiac Function by Enhancing the Expression of SIRT3 and PGC-1α in a Rat Model of Isoproterenol-Induced Cardiomyopathy.

Mitochondrial biosynthesis regulated by the PGC-1α-NRF1-TFAM pathway is considered a novel potential therapeutic target to treat heart failure (HF). Perindopril (PER) is an angiotensin-converting enzyme inhibitor that has proven efficacy in the prevention of HF; however, its mechanism is not well established. In this study, to investigate the mechanisms of PER in cardiac protection, a rat model of cardiomyopathy was established by continuous isoproterenol (ISO) stimulation. Changes in the body weight, heart weight index, echocardiography, histological staining, mitochondrial microstructure, and biochemical indicators were examined. Our results demonstrate that PER reduced myocardial remodeling, inhibited deterioration of cardiac function, and delayed HF onset in rats with ISO-induced cardiomyopathy. PER markedly reduced reactive oxygen species (ROS) production, increased the levels of antioxidant enzymes, inhibited mitochondrial structural destruction and increases the number of mitochondria, improved the function of the mitochondrial respiratory chain, and promoted ATP production in myocardial tissues. In addition, PER inhibited cytochrome C release in mitochondria and caspase-3 activation in the cytosol, thereby reducing the apoptosis of myocardial cells. Notably, PER remarkably up-regulated the mRNA and protein expression levels of Sirtuin 3 (SIRT3), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (TFAM) in myocardial cells. Collectively, our results suggest that PER induces mitochondrial biosynthesis-mediated enhancement of SIRT3 and PGC-1α expression, thereby improving the cardiac function in rats with ISO-induced cardiomyopathy.

Non-HDL-c/TC: A Novel Lipid-Related Marker in the Assessment of Severity of Coronary Artery Lesions and Cardiovascular Outcomes.

BACKGROUND: Non-high-density lipoprotein cholesterol (non-HDL-c) predicts the severity of coronary artery lesions in patients not treated with statin. The association between non-HDL-c and severity of coronary artery lesions in patients treated with lipid-lowering therapy has been unknown. HYPOTHESIS: We hypothesize a novel marker of non-HDL-c/TC predicts the severity of coronary artery lesions and clinical outcomes in 12 months in the patients treated with statin. METHOD: 473 subjects who met inclusion criteria were eligible for inclusion. Coronary artery angiography (CAG) was performed, and the Gensini score (GS) was calculated in all the subjects divided into three subgroups of low risk, medium risk, and high risk by the tertiles of GS. The non-HDL-c value was calculated as TC minus HDL-c, while non-HDL-c/TC was the ratio of non-HDL-c and TC. RESULTS: The concentration of non-LDL-c differed between non-obstructive-CAD group and obstructive-CAD group (P < 0.05), and non-HDL-c/TC was elevated in the obstructive-CAD group (P < 0.05). Increased GS was associated with increasing non-HDL-c/TC (P < 0.05). Non-HDL-c/TC (OR: 108.50, 95% CI: 1.57-7520.28; P=0.030) remained as an independent predicting factor of high risk under GS stratification. In unadjusted Cox model, high non-HDL-c/TC (RR: 1.976, 95% CI: 1.155-3.382; P=0.013) predicted the occurrence of adverse events. After multivariate adjustment, high non-HDL-c/TC (RR: 1.921, 95% CI: 1.105-3.339; P=0.021) was an independent predictor of poor outcomes. CONCLUSION: High level of non-HDL-c/TC presented an excellent prognostic value compared with other lipid-related markers in CAD patients treated with statin.

Self-assembled GFFYK peptide hydrogel enhances the therapeutic efficacy of mesenchymal stem cells in a mouse hindlimb ischemia model.

Mesenchymal stem cell (MSC) transplantation has emerged as a very promising strategy for the treatments of peripheral artery disease (PAD). However, MSC-based therapies are limited by low cell retention and survival rate in the ischemic zone. Small molecular (SM) hydrogels have shown attractive abilities to enhance the therapeutic effects of human MSCs via promoting their proliferation or maintaining their differentiation potential. Here, we designed and synthesized a new bioactive and biocompatible hydrogel, Nap-GFFYK-Thiol, using disulfide bonds as cleavable linkers to control the molecular self-assembly and we hypothesized this hydrogel could enhance the retention and engraftment of human placenta-derived MSCs (hP-MSCs) in a mouse ischemic hindlimb model. In vitro results demonstrated that the Nap-GFFYK-Thiol hydrogel increased cell viability through paracrine effects. Moreover, it enhanced the proangiogenic and anti-apoptotic effects of hP-MSCs. In vivo, Nap-GFFYK-Thiol hydrogel improved the hP-MSC retention in the murine ischemic hindlimb model as visualized by bioluminescence imaging. Furthermore, cotransplantation of hP-MSCs with hydrogel improved blood perfusion, leading to superior limb salvage. These therapeutic effects may attribute to reduced inflammatory cell infiltration, enhanced angiogenesis as well as suppressed collagen deposition. In conclusion, the Nap-GFFYK-Thiol hydrogel fabricated using disulfide bonds as cleavable linkers serves as an artificial niche for promoting hP-MSC survival and proangiogenic factor secretion in PAD therapy and thereby provide an alternative strategy for PAD therapy. STATEMENT OF SIGNIFICANCE: Although several phase I/II clinical trials of MSC-based treatments for critical limb ischemia (CLI) are ongoing, MSC-based therapies are still challenged by the low quality and quantity of cells in the ischemic zone, especially in cases of extensive or irreversible damage. Hydrogels have favorable biocompatibility and safety records in the medical field. In the current study, we engineered a new bioactive and biocompatible hydrogel, Nap-GFFYK-Thiol, using disulfide bonds as cleavable linkers to enhance the therapeutic efficacy of human placenta-derived MSCs (hP-MSCs) in mouse limb ischemia model. Notably, Nap-GFFYK-Thiol hydrogel acts as an artificial niche for promoting hP-MSC survival and proangiogenic factor secretion in PAD therapy, which further promoted the restoration of blood perfusion and regeneration of muscle cells. Considering the proangiogenic effect of Nap-GFFYK-Thiol on hP-MSCs, our results may provide a new strategy for the treatment of PAD.

Tongmai Yangxin pills anti-oxidative stress alleviates cisplatin-induced cardiotoxicity: Network pharmacology analysis and experimental evidence.

BACKGROUND: Tongmai Yangxin Pills (TMYXP), a traditional Chinese patent medicine, has been widely used to treat coronary heart disease for few decades. However, the potential protective effect of TMYXP on cisplatin-induced cardiotoxicity has not been reported. METHODS: The target proteins corresponding to compounds from Pharmmapper database, PubMed database and ChEMBL database were collected and construct a 'TMYXP-compound-target' network. DAVID database was used for annotation and enrichment pathways and String 9.1 database was used for analysis the protein-protein interaction. Cisplatin-induced rat cardiotoxicity model was established to verify the protective effects mechanism of TMYXP. RESULTS: The target proteins corresponding to compounds from multiple databases were collected and construct a TCM-compound-target network to enriched pathways with high enrichment score. GO analysis and enrichment clusters point that response of oxidative stress is the main biological process of TMYXP, and Nrf2 signaling pathway and MAPK signaling pathway might be the key functional pathways. In vivo experiments, we proved that TMYXP improves anti-oxidative stress ability and reduce apoptosis through regulating Nrf2/HO-1 pathway and p38-MAPK pathway. CONCLUSION: The effects of TMYXP on regulate cardiomyocyte free radical balance and reduce apoptosis, making it possible as a drug candidate for platinum chemotherapeutic induced cardiac injury.

Differential and Predictive Value of Galectin-3 and Soluble Suppression of Tumorigenicity-2 (sST2) in Heart Failure with Preserved Ejection Fraction.

BACKGROUND Galectin-3 and soluble suppression of tumorigenicity-2 (sST2) are promising biomarkers of cardiac fibrosis and ventricular remodeling. The purpose of this study was to investigate the diagnostic and predictive value of galectin-3 and sST2 for use in patients who have heart failure with preserved ejection fraction (HFpEF). MATERIAL AND METHODS A total of 217 hospitalized patients with HF and 30 controls from a physical examination center were included. Venous blood was collected for the detection of circulating expression of galectin-3 and sST2. All the included patients were followed up regularly for 1 year (12±1 months). RESULTS The concentrations of galectin-3 and NT-proBNP were substantially higher following decreased ejection fraction (both P=0.000), except for sST2 (P=0.068 vs. control). In ROC analyses, galectin-3 and NT-proBNP distinguished HFpEF from controls with an area under the curve (AUC) of 0.819 (95% CI: 0.75-0.89, P=0.000) and 0.806 (95% CI: 0.66-0.82, P=0.000). In contrast, sST2 obtained a lower AUC of 0.584 (95% CI: 0.49-0.68, P=0.17) compared to galectni-3 and NT-proBNP. After adjustment for clinical factors and NT-proBNP, galectin-3 was strongly correlated with an increased risk of the endpoint events in HFpEF patients, and the hazard ratio per 1 SD increase of the galectin-3 level was 2.33 (95%CI: 1.72-2.94, P=0.009). CONCLUSIONS Galectin-3 is superior to sST2 in distinguishing HFpEF from controls and HFrEF.

Additional Diagnostic Value of Growth Differentiation Factor-15 (GDF-15) to N-Terminal B-Type Natriuretic Peptide (NT-proBNP) in Patients with Different Stages of Heart Failure.

BACKGROUND Growth differentiation factor-15 (GDF-15) is a promising biomarker of cardiac remodeling. The purpose of this study was to explore the diagnostic value of plasma GDF-15 levels in different stages of heart failure (HF) and to assess the relationship with ventricular remodeling. MATERIAL AND METHODS We enrolled 219 HF patients from the Department of Cardiology in Tianjin Union Medical Center as the HF group and 32 healthy subjects as the control group. Circulating GDF-15, NT-proBNP, procollagen I C-terminal propeptide (PICP), and N-terminal procollagen III propeptide (PIIINP) levels were measured using ELISA. Associations between GDF-15 and clinical indicators in cardiac remodeling were assessed using receiver operating characteristic (ROC) curves and Spearman correlation. All the patients were followed up for 1 year. RESULTS The level of plasma GDF-15 in HF patients was higher than in the control group (P<0.05) and increased with higher ACCF/AHA and NYHA classification (P<0.05). Patients with HFrEF had higher GDF-15 levels compared to patients with HFmrEF (P<0.05). GDF-15 and left ventricular mass index (LVMI) were significantly increased as early as the pre-clinical HF stage. Also, GDF-15 levels were positively correlated to LVMI (r=0.433, P<0.05), PICP (r=0.378, P<0.001) and PIIINP (r=0.382, P<0.001). ROC curves were constructed and GDF-15 plus NT-proBNP (AUC=0.905, 95%CI: 0.868-0.942, P<0.001) was superior to NT-proBNP (AUC=0.869, 95%CI: 0.825-0.913, P<0.001) in identifying HF. GDF-15 levels did not predict prognosis after a 1-year follow-up period. CONCLUSIONS GDF-15 combined with NT-proBNP significantly improves the accuracy of diagnosing HF. Plasma GDF-15 levels can indirectly reflect the degree of cardiac remodeling and fibrosis.

Prognostic value of sST2 and NT-proBNP at admission in heart failure with preserved, mid-ranged and reduced ejection fraction.

OBJECTIVE: The aim of this study was to compare sST2 at admission with N-terminal pro B-type natriuretic peptide (NT-proBNP) in prognostic accuracy among different types of heart failure (HF) and clarifying the enhanced prognostic value in patients with HF by the combination of the two biomarkers. METHODS AND RESULTS: A total of 164 consecutive patients admitted to our institution for HF were divided into three groups of HF with reduced ejection fraction (HFrEF), HF with mid-range ejection fraction (HFmrEF) and HF with preserved ejection fraction (HFpEF). Circulating sST2 and NT-proBNP were measured using the enzyme-linked immunosorbent assay (ELISA). The sST2 level was only significantly higher in HFrEF when compared with HFpEF. At ROC analysis to one-year adverse events, only sST2 showed predictive value in HFmrEF with an optimal cut-off value of 147.66 ng/ml (AUC 0.697, p = .045, sensitivity 75%, specificity 75.8%), while both NT-proBNP and sST2 showed a significant predictive value in HFpEF (p = .036 vs .042; AUC 0.683, sensitivity 71.4%, specificity 55.6%; AUC 0.677, sensitivity 64.3%, specificity 83.3%) with an optimal cut-off value of 1054.50 pg/ml and 117.80 ng/ml. Multivariate regression analysis suggested that sST2 and NT-proBNP could be biomarkers for predicting 1-year adverse events of HF (OR = 4.384, 95% CI = 1.661-11.570 vs. OR = 3.451, 95% CI = 1.254-9.497). Adverse events occurred frequently within one year both in sST2 and in NT-proBNP above the median. CONCLUSIONS: sST2 can provide different prognostic information in distinct types of HF, and even be superior to NT-proBNP. sST2 combined with NT-proBNP can improve predicting accuracy.

Increased complements and high-sensitivity C-reactive protein predict heart failure in acute myocardial infarction.

The aim of the present study was to investigate whether the serum levels of complements and high-sensitivity C-reactive protein (hs-CRP) in patients with acute myocardial infarction (AMI) are associated with the severity of myocardial injury. Consecutive patients (n=110) with AMI and 33 healthy individuals, who served as control subjects, were enrolled from May 2013 to February 2015. These patients were divided into two groups, those with ST segment elevation MI (STEMI) and those with non-ST segment elevation MI (NSTEMI). The patients with STEMI exhibited progression to diastolic dysfunction and heart failure. Furthermore, the results revealed that the level of serum complement and hs-CRP in patients with AMI increased rapidly when compared with the subjects from the control group, particularly in the STEMI patients, at different time-points. A statistically significant elevation of the complement and hs-CRP levels was observed at day 3 after AMI in the STEMI group. The activation of complement and hs-CRP following AMI may serve as a specific marker to successfully predict left ventricular dysfunction. Thus, biomarker-based approaches may be adopted to identify the severity of AMI with distinct pathophysiologic responses in order to rationally implement clinical therapeutic strategies.