Arginine vasopressin induces ferroptosis to promote heart failure via activation of the V1aR/CaN/NFATC3 pathway.

Arginine vasopressin (AVP) is a key contributor to heart failure (HF), but the underlying mechanisms remain unclear. In the present study, a mouse model of HF and human cardiomyocyte (HCM) cells treated with dDAVP are generated in vivo and in vitro, respectively. Hematoxylin and eosin (HE) staining is used to evaluate the morphological changes in the myocardial tissues. A colorimetric method is used to measure the iron concentration, Fe 2+ concentration and malondialdehyde (MDA) level. Western blot analysis is used to examine the protein levels of the V1a receptor (V1aR), calcineurin (CaN), nuclear factor of activated T cells isoform C3 (NFATC3), glutathione peroxidase 4 (GPX4) and acyl-CoA synthase long chain family member 4 (ACSL4). Immunoprecipitation (IP) and luciferase reporter assays are performed to determine the interaction between NFATC3 and ACSL4. Both in vivo and in vitro experiments reveal that the V1aR-CaN-NFATC3 signaling pathway and ferroptosis are upregulated in HFs, which are verified by the elevated protein levels of V1aR, CaN, NFATC3 and ACSL4; reduced GPX4 protein level; and enhanced Fe 2+ and MDA levels. We further find that inhibiting NFATC3 by suppressing the V1aR/CaN/NFATC3 pathway via V1aR/CaN inhibitors or sh-NFATC3 not only alleviates HF but also inhibits AVP-induced ferroptosis. Mechanistically, sh-NFATC3 significantly reverses the increase in AVP-induced ACSL4 protein level, Fe 2+ concentration, and MDA level by directly interacting with ACSL4. Our results demonstrate that AVP enhances ACSL4 expression by activating the V1aR/CaN/NFATC3 pathway to induce ferroptosis, thus contributing to HF. This study may lead to the proposal of a novel therapeutic strategy for HF.

Influence of enhanced external counterpulsation on endothelial function: a meta-analysis of randomized controlled trials.

OBJECTIVES: Enhanced external counterpulsation (EECP) is an effective and noninvasive treatment for patients with refractory angina and chronic heart failure. However, previous studies evaluating the influence of EECP on endothelial function showed inconsistent results. This systematic review and meta-analysis was conducted to evaluate the effects of EECP on endothelial function measured by brachial artery flow-mediated dilation (FMD). DESIGN: PubMed, Embase, Cochrane Library, CNKI, and Wanfang databases were searched for randomized controlled trials comparing the influence of EECP versus usual care on FMD in adult population. A random-effects model incorporating the potential influence of heterogeneity was used to pool the results. RESULTS: Nineteen studies with 1647 patients were included in the meta-analysis. Compared with usual care or conventional therapy, additional treatment with EECP for 3-7 weeks was associated with a significantly improved FMD (mean difference [MD]: 1.96%, 95% confidence interval [CI]: 1.57-2.36, p < 0.001, I2 = 52%). Subgroup analysis showed consistent results in patients with coronary artery disease and in patients with other diseases (p for subgroup difference = 0.21). Results of meta-regression analysis showed that the mean baseline FMD level was positively correlated with the influence of EECP on FMD (coefficient = 0.42, p < 0.001). Results of subgroup analysis suggested that the increment of FMD following EECP was larger in patients with baseline FMD ≥ 5% (MD: 2.69, 95% CI: 2.27-3.10, p < 0.001; I2 = 15%) compared to those with baseline FMD < 5% (MD: 1.49, 95% CI: 1.13-1.85, p < 0.001; I2 = 0%; p for subgroup difference < 0.001). CONCLUSIONS: EECP may be effective in improving endothelial function measured by FMD.

Role and mechanism of the lncRNA SNHG1/miR­450b­5p/IGF1 axis in the regulation of myocardial ischemia reperfusion injury.

The increasing rates of morbidity and mortality caused by ischemic heart disease pose a serious threat to human health. Long non­coding (lnc)RNA small nucleolar RNA host gene 1 (SNHG1) has a protective effect on the myocardium. In the present study, the role of lncRNA SNHG1 in myocardial ischemia reperfusion injury (MIRI) and the underlying mechanisms were investigated. After hypoxia/reoxygenation (H/R) induction, the expression levels of lncRNA SNHG1 were detected using reverse transcription­quantitative PCR. After lncRNA SNHG1 overexpression via cell transfection, cell viability was detected using an MTT assay, apoptotic rates were detected using TUNEL staining, apoptosis­related protein expression levels were detected using western blotting and respective kits were used to measure the oxidative stress levels. The Encyclopedia of RNA Interactomes database predicted the presence of binding sites between lncRNA SNHG1 and microRNA (miR)­450b­5p, and between miR­450b­5p and insulin­like growth factor 1 (IGF1). These interactions were then verified using luciferase reporter assays. Subsequently, the regulatory mechanism underlying the lncRNA SNHG1/miR­450b­5p/IGF1 axis in MIRI was investigated by overexpressing miR­450b­5p and knocking down IGF1 expression in H/R­induced cells. Finally, the expression of PI3K/Akt signaling pathway­related proteins was detected using western blotting. lncRNA SNHG1 expression was significantly downregulated in H/R­induced AC16 cells. lncRNA SNHG1 overexpression significantly inhibited apoptosis and decreased oxidative stress levels in H/R­induced AC16 cells, which was mediated via regulation of the miR­450b­5p/IGF1 axis and activation of the PI3K/Akt signaling pathway. Therefore, the present study suggested that activation of the PI3K/Akt signaling pathway via the lncRNA SNHG1/miR­450b­5p/IGF1 axis inhibited the apoptosis and oxidative stress levels of H/R­induced AC16 cells.

Effect of chromium supplementation on hs-CRP, TNF-α and IL-6 as risk factor for cardiovascular diseases: A meta-analysis of randomized-controlled trials.

BACKGROUND: The objective of this systematic review is to assess the relationship between chromium supplementation and inflammatory biomarkers levels (hs-CRP, TNF-α, IL-6) as risk factor for cardiovascular diseases. Recent studies raise questions regarding the potential of chromium supplementation to decrease the blood-levels of inflammatory markers, lowering cellular oxidative stress as markers of myocardial infarction; however, the results of the researches are inconclusive. METHODS: The following databases including PubMed, Scopus, Cochran Library and Embase databases were systematically searched until April 2020. Analysis was performed using random-effect model. RESULTS: The pooled findings for biomarkers of inflammation showed that chromium supplementation significantly reduced serum high-sensitivity C-reactive protein (hs-CRP) (WMD: -0.87 mg/dL, 95% CI: -1.49, -0.26), and tumor necrosis factor-alpha (TNF-α) (WMD: -0.97 pg/ml; 95% CI: -1.92, -0.01) and chromium insignificantly reduced interleukin -6 (IL-6) (WMD: -0.45 pg/ml, 95% CI: -1.18, 0.29). CONCLUSION: Overall, the results of this systematic review and meta-analysis imply that chromium supplementation may help to improve biomarkers of inflammation as markers of myocardial infarction.

lncRNA ZFAS1 promotes ox-LDL induced EndMT through miR-150-5p/Notch3 signaling axis.

EndMT is an active contributor to atherosclerosis pathology, and lncRNAs is widely involved in the occurrence and development of atherosclerosis. The purpose of this study was to investigate the regulatory mechanisms of ZFAS1 in EndMT of atherosclerosis. Here, the ApoE-/- mice were feed with high-fat diet to establish the atherosclerosis model, and HUVECs was stimulated with ox-LDL to induce EndMT. RT-PCR and western blot were used to detect the mRNA and protein expression, respectively. The expression of EndMT markers were detected by immune-fluorescence. The relationships among ZFAS1, miR-150-5p and Notch3 were evaluated by luciferase reporter assay. The role of ZFAS1 in EndMT and its dependence on miR-150-5p/Notch3 axis was further detected by knocking down or over-expressing ZFAS1. We found that ZFAS1 and Notch3 were upregulated while miR-150-5p was downregulated in atherosclerosis mice and ox-LDL-treated HUVECs. The expression of CD31 and vWF were significant decreased, while the α-SMA and vimentin were significant increased in ox-LDL-treated HUVECs, and overexpression of ZFAS1 enhanced the effect of ox-LDL on HUVECs. Further, ZFAS1 functions as a ceRNA to increase Notch3 expression through sponging miR-150-5p, and miR-150-5p mimic or si-Notch3 could reverse LV-ZFAS1-mediated EndMT. In summary, lncRNA ZFAS1 promotes ox-LDL induced HUVECs EndMT through regulating miR-150-5p/Notch3 axis.

Interference of TGF-ß1/Smad7 signal pathway affects myocardial fibrosis in hypertension.

Hypertension can cause myocardial fibrosis, during which tumor growth factor-beta 1 (TGF-ß1) can facilitate myocardial cell proliferation and transition towards myofibroblast (MFB). Smad7 is a negative regulator of TGF-ß1/Smads signal pathway. This study used hypertension rat model to investigate the regulatory role of TGF-ß1/Smad7 signal pathway in myocardial fibrosis. Rat renal hypertension model was established to test collagen volume fraction, myocardial hydroxyl proline content and COL1A1 expression as well as the expression of TGF-ß1 and Smad7. The expressions of TGF-ß1, Smad7, COLA1A1 and α-SMA at certain generations (P2, P4 and P6) were measured in cultured human cardiac fibroblast (HCF) during differentiation towards MFB differentiation. P6 generation HCF was transfected with pIRES2-EGFP-Smad7 and pIRES2-EGFP-blank followed by measuring expressions of TGF-ß1, Smad7, COLA1A1 and α-SMA. Hydroxyl-proline content and collagen volume fraction were compared between Ad-NC and Ad-Smad7 injection groups. Hypertensive rats had significantly elevated collagen volume fraction, hydroxyl proline contents, and expression of COLA1 and TGF-ß1 than Sham group, whilst Smad7 expression was lower. With increased cell passage, HCF showed gradually increased TGF-ß1, COL1A1 and α-SMA expression, plus decreased Smad7 expression. Over-expression of Smad7 remarkably decreased COLA1 and α-SMA expression in HCF. Tail vein injection of Ad-Smad7 decreased both hydroxyl proline and collagen volume fraction. Elevated TGF-ß1 expression and decreased Smad7 expression are found in fibrotic myocardial tissues of hypertensive rats. Over-expression of Smad7 inhibits differentiation of HCF towards MFB cells, thus decreasing myocardial fibrosis in hypertensive rats.

Development and validation of T-ARMS-PCR to detect CYP2C19*17 allele.

BACKGROUND: CYP2C19*17 (rs12248560) is a functional single nucleotide polymorphism (SNP) in the CYP2C19 gene. It has been shown that CYP2C19*17 is associated with the clinical outcome of some drugs metabolized by CYP2C19 and a decreased risk of some diseases. The aim of this study was to develop a reliable and simple method to detect this polymorphism. METHODS: Tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) was used to detect the CYP2C19*17 polymorphism. A total of 93 samples were screened by this method, and the results of T-ARMS-PCR were validated by DNA sequencing. RESULTS: There were 91 samples with the CC genotype (97.8%) and two samples with the CT genotype (2.2%). The frequency of the C allele was 98.9%, and the frequency of the T allele was 1.1%. The DNA sequencing results were completely concordant with the T-ARMS-PCR results. CONCLUSION: T-ARMS-PCR can detect the CYP2C19*17 polymorphism with high accuracy, low costs, and a simple process.

[Effects of Tongxinluo Capsule on Platelet Activating Factor, Vascular Endothelial Function, Blood Flow of Thrombolysis in Myocardial Infarction in Acute Myocardial Infarction Patients after Delayed Percutaneous Coronary Intervention].

OBJECTIVE: To explore effects of Tongxinluo Capsule (TC) on platelet activating factor (PAF), vascular endothelial function, thrombolysis in myocardial infarction (TIMI) blood flow, and heart function in acute myocardial infarction (AMI) patients after delayed percutaneous coronary intervention (PCI). METHODS: Totally 80 AMI inpatients were recruited at Department of Cardiology, People's Hospital of Jiangxi Province, from Jan. 2008 to Sep.2013. Those in line with inclusion criteria were randomly assigned to TC treatment group and the conventional treatment group by random digit table, 40 in each group. Besides, another 40 healthy subjects from examinees at Outpatient Department were recruited as a healthy control group. PCI was performed after 1-week treatment. Then blood samples were collected, and then blood contents of CD62P, CD63, GP II b/III a, ET-1, NO, and plasma von Willebrand factor (vWF) levels were detected. Coronary TIMI blood flow and corrected TIMI frame count (CTFC) were determined during PCI. Meanwhile, noninvasive blood pressure (BP) and heart rate (HR) were recorded before and after PCI, and cardiac function measured. They were compared with the healty control group. RESULTS: Compared with the healthy control group, blood contents of CD62p, CD63, GP II b/IIIa receptor compound, vWF, and ET-1 significantly increased, but NO significantly decreased in AMI patients (all P < 0.05). After 1-week intervention of TC, blood contents of CD62p, CD63, GP II b/IIIa receptor compound, vWF, NO, and ET-1 significantly decreased (P < 0.05, P < 0.01). Compared with the conventional treatment group at the same time point, blood contents of CD62p, CD63, GP II b/IIIa receptor compound, vWF, and ET-1 decreased more significantly in the TC group (P < 0.05, P < 0.01), increased NO levels were also more obviously seen (P < 0.01). The aforesaid parameters changed more obviously at day 30, as compared with those changes at week 1 (P < 0.05, P < 0.01). The TIMI blood flow grade and CTFC were more obviously improved after PCI in the two treatment groups. Better TIMI blood flow was seen in the TC group. TIMI level 3 blood flow rate was higher in the TC group than in the conventional treatment group with statistical difference (P < 0.05). The left ventricular ejective factor (LVEF) after PCI was obviously elevated in the TC group and the conventional treatment group (P < 0.01), and the improvement was more obviously seen in the TC group (P < 0.05). There were 6 cases of recurrent angina, 3 cases of ventricular tachycardial (VT)/ventricular fibrillation (VF), 6 cases of heart failure (HF), 1 case of cardiac sudden death in the conventional treatment group, with the total incidence of cardiovascular events being 40% (16/40). There were 2 cases of recurrent angina, 2 cases of VT/VF, 2 cases of HF, no cardiac sudden death in the TC treatment group, with the total incidence of cardiovascular events being 15% (6/40). There was statistical difference in the recurrent rate of cardiovascular events between the two groups (χ² = 2.27, P < 0.05). CONCLUSION: TC not only could prevent coronary embolism of AMI patients after delayed PCI, attenuate vascular endothelial injury, but also could improve TIMI blood flow, and strengthen cardiac systolic function.

Application of percutaneous balloon mitral valvuloplasty in patients of rheumatic heart disease mitral stenosis combined with tricuspid regurgitation.

BACKGROUND: Tricuspid regurgitation (TR) is frequently associated with severe mitral stenosis (MS), the importance of significant TR was often neglected. However, TR influences the outcome of patients. The aim of this study was to investigate the efficacy and safety of percutaneous balloon mitral valvuloplasty (PBMV) procedure in rheumatic heart disease patients with mitral valve (MV) stenosis and tricuspid valve regurgitation. METHODS: Two hundred and twenty patients were enrolled in this study due to rheumatic heart disease with MS combined with TR. Mitral balloon catheter made in China was used to expand MV. The following parameters were measured before and after PBMV: MV area (MVA), TR area (TRA), atrial pressure and diameter, and pulmonary artery pressure (PAP). The patients were followed for 6 months to 9 years. RESULTS: After PBMV, the MVAs increased significantly (1.7 ± 0.3 cm 2 vs. 0.9 ± 0.3 cm 2 , P < 0.01); TRA significantly decreased (6.3 ± 1.7 cm 2 vs. 14.2 ± 6.5 cm 2 , P < 0.01), right atrial area (RAA) decreased significantly (21.5 ± 4.5 cm 2 vs. 25.4 ± 4.3 cm 2 , P < 0.05), TRA/RAA (%) decreased significantly (29.3 ± 3.2% vs. 44.2 ± 3.6%, P < 0.01). TR velocity (TRV) and TR continue time (TRT) as well as TRV × TRT decreased significantly (183.4 ± 9.4 cm/s vs. 254.5 ± 10.7 cm/s, P < 0.01; 185.7 ± 13.6 ms vs. 238.6 ± 11.3 ms, P < 0.01; 34.2 ± 5.6 cm vs. 60.7 ± 8.5 cm, P < 0.01, respectively). The postoperative left atrial diameter (LAD) significantly reduced (41.3 ± 6.2 mm vs. 49.8 ± 6.8 mm, P < 0.01) and the postoperative right atrial diameter (RAD) significantly reduced (28.7 ± 5.6 mm vs. 46.5 ± 6.3 mm, P < 0.01); the postoperative left atrium pressure significantly reduced (15.6 ± 6.1 mmHg vs. 26.5 ± 6.6 mmHg, P < 0.01), the postoperative right atrial pressure decreased significantly (13.2 ± 2.4 mmHg vs. 18.5 ± 4.3 mmHg, P < 0.01). The pulmonary arterial pressure decreased significantly after PBMV (48.2 ± 10.3 mmHg vs. 60.6 ± 15.5 mmHg, P < 0.01). The symptom of chest tightness and short of breath obviously alleviated. All cases followed-up for 6 months to 9 years (average 75 ± 32 months), 2 patients with severe regurgitation died (1 case of massive cerebral infarction, and 1 case of heart failure after 6 years and 8 years, respectively), 2 cases lost access. At the end of follow-up, MVA has been reduced compared with the postoperative (1.4 ± 0.4 cm 2 vs. 1.7 ± 0.3 cm 2 , P < 0.05); LAD slightly increased compared with the postoperative (45.2 ± 5.7 mm vs. 41.4 ± 6.3 mm, P < 0.05), RAD slightly also increased compared with the postoperative (36.1 ± 6.3 mm vs. 28.6 ± 5.5 mm, P < 0.05), but did not recover to the preoperative level. TRA slightly increased compared with the postoperative, but the difference was not statistically significant (P > 0.05). The PAP and left ventricular ejection fraction appeared no statistical difference compared with the postoperative (P > 0.05), the remaining patients without serious complications. CONCLUSIONS: PBMV is a safe and effective procedure for MS combined with TR in patients of rheumatic heart disease. It can alleviate the symptoms and reduce the size of TR. It can also improve the quality-of-life and prognosis. Its recent and mid-term efficacy is certain. While its long-term efficacy remains to be observed.

Application of peripheral-blood-derived endothelial progenitor cell for treating ischemia-reperfusion injury and infarction: a preclinical study in rat models.

BACKGROUND: Our aim was to explore the therapeutic effects of peripheral blood-derived endothelial progenitor cells (PB-EPC) in cardiac ischemia-reperfusion infarction models in rats and in in vitro culture systems. METHODS: Rat models of ischemia reperfusion and myocardial infarction were developed using male, Sprague-Dawley rats. Cardiomyocyte and endothelial cell cultures were also established. Therapeutic effects of PB-EPCs were examined in vivo and in vitro in both models. Rats underwent either cardiac ischemia-reperfusion (n = 40) or infarction (n = 56) surgeries and were transplanted with genetically modified EPCs. Treatment efficacy in the ischemia-reperfusion group was measured by infarct size, myocardial contraction velocity, and myeloperoxidase activity after transplantation. Cardiomyocyte survival and endothelial cell apoptosis were investigated in vitro. Vascular growth-associated protein expression and cardiac function were evaluated in the myocardial infarction group by western blot and echocardiography, respectively. RESULTS: Infarct size and myeloperoxidase activity were significantly decreased in the ischemia-reperfusion group, whereas myocardial contractility was significantly increased in the EPC and Tß4 groups compared with that in the control group. In contrast, no differences were found between EPC + shRNA Tß4 and control groups. Rates of cardiomyocyte survival and endothelial cell apoptosis were significantly higher and lower, respectively, in the EPC and Tß4 groups than in the control group, whereas no differences were found between the EPC + shRNA Tß4 and control group. Four weeks after myocardial infarction, cardiac function was significantly better in the EPC group than in the control group. Expressions of PDGF, VEGF, and Flk-1 were significantly higher in EPC group than in control group. CONCLUSIONS: Study findings suggest that PB-EPCs are able to protect cardiomyocytes from ischemia-reperfusion or infarction-induced damage via a Tß4-mediated mechanism. EPCs may also provide protection through increased expression of proteins involved in mediating vascular growth. Autologous peripheral-blood-derived EPCs are readily available for efficient therapeutic use without the concerns of graft rejection.

[Comparative study of interventricular electrical and mechanical synchrony at different cardiac pacing sites].

OBJECTIVE: To compare the different impacts of right ventricular apex, right ventricular outflow tract septum and left ventricular outflow tract septum region on interventricular electro-mechanical synchronization and assess the ideal pacing sites for maintaining the interventricular electro-mechanical synchronization. METHODS: A total of 30 patients without organic heart disease were operated with radiofrequency ablation at our hospital. The mapping electrodes were implanted post-operatively on the left ventricular posterior wall (LVPLW) and right ventricular anterior lateral wall (RVALW) respectively. And the ablation electrodes were placed subsequently in right ventricular apex, right ventricular outflow tract septum region and left ventricular outflow tract septum. The difference values were measured between transmission time from pacemaker to LVPLW, from pacemaker to RVALW and between aortic pre-ejection interval (APEI) and pulmonary artery pre-ejection interval (PPEI). Then their correlations were compared. RESULTS: When pacing at right ventricular apex, the difference value between transmission time from pacemaker to LVPLW and from pacemaker to RVALW was (34 ± 7) ms. And it was (18 ± 4) ms while pacing at right ventricular outflow tract septum region and (12 ± 4) ms at left ventricular outflow tract septum region. There was significant difference (P < 0.01). The absolute value of APEI-PPEI was (25 ± 5) ms at right ventricular apex, (13 ± 4) ms at right ventricular outflow tract septum region and (11 ± 3) ms at left ventricular outflow tract septum region. And there was significant difference (P < 0.01). The absolute value of APEI-PPEI was positively correlated with the change of LVPLW-RVALW (r = 0.993, P < 0.01). Left ventricular outflow tract septum pacing showed ABp and left ventricle end-systolic pressure significantly increased [(127 ± 23) mm Hg, (142 ± 22) mm Hg, P < 0.05], left ventricular end-diastolic pressure was significantly lower [(9 ± 3) mm Hg, P < 0.05]. CONCLUSION: Compared with right ventricular apical pacing and right ventricular outflow tract ventricular septal pacing, left ventricular outflow tract septum has a smaller impact on the electro-mechanical synchronization. It conforms more closely to the physiological pacing so that there is a higher synchronization of electrical and mechanical ventricular contractions.

[Percutaneous coronary intervention of patients with complex small coronary artery lesion unsuitable for coronary artery bypass graft].

OBJECTIVE: To investigate the safety and efficacy of percutaneous coronary intervention (PCI) in patients with low left ventricular ejection fraction (LVEF) and complex small coronary artery lesions. METHODS: Complete or partial post-PCI revascularization of coronary artery was employed in 16 patients with a low LVEF and complex small coronary artery lesions who were unsuitable for CABG (coronary artery bypass grafting). All cases were observed with regards to immediate success rate of operation, complication, hospitalization duration, improvement of cardiac function and LVEF and major adverse cardiac events (including cardiac death, myocardial infarction and target lesion revascularization) at 12 months post-operation. RESULTS: All cases were successfully treated without death and severe complications while the hospitalization duration was (11 +/- 5) days. The follow-up survey at 12 months post-operation showed that no major adverse cardiac event occurred, the post-operative improvement of cardiac function was from III - IV grade to I - II grade, the improvement of LVEF was from 25% - 45% [(29 +/- 8)%] to 32% - 48% [(37 +/- 7)%], left ventricular end diastolic diameter (LVDd) was shortened from 52 - 79 (66 +/- 11) mm to 49 - 68 (58 +/- 8) mm. The reexamination of 14 cases by coronary angiography at 12 months post-operation showed that there was no intra-stent thrombosis while 20% - 40% intra-stent restenosis occurred in 2 cases. CONCLUSION: For patients with a low LVEF and complex small coronary artery lesions, PCI is a safe and effective method to lower the mortality rate of CHD patients with heart failure and improve the long-term patient prognosis.