Puerarin alleviates atherosclerosis via the inhibition of Prevotella copri and its trimethylamine production.

OBJECTIVE: Puerarin (PU) is a natural compound that exhibits limited oral bioavailability but has shown promise in the treatment of atherosclerosis (AS). However, the precise mechanisms underlying its therapeutic effects remain incompletely understood. This study aimed to investigate the effects of PU and its mechanisms in mitigating AS in both mice and humans. DESIGN: The impact of PU on AS was examined in ApoE -/- mice fed a high-fat diet (HFD) and in human patients with carotid artery plaque. To explore the causal link between PU-associated gut microbiota and AS, faecal microbiota transplantation (FMT) and mono-colonisation of mice with Prevotella copri (P. copri) were employed. RESULTS: PU alleviated AS by modulating the gut microbiota, as evidenced by alterations in gut microbiota composition and the amelioration of AS following FMT from PU-treated mice into ApoE-/- mice fed HFD. Specifically, PU reduced the abundance of P. copri, which exacerbated AS by producing trimethylamine (TMA). Prolonged mono-colonisation of P. copri undermines the beneficial effects of PU on AS. In clinical, the plaque scores of AS patients were positively correlated with the abundance of P. copri and plasma trimethylamine-N-oxide (TMAO) levels. A 1-week oral intervention with PU effectively decreased P. copri levels and reduced TMAO concentrations in patients with carotid artery plaque. CONCLUSION: PU may provide therapeutic benefits in combating AS by targeting P. copri and its production of TMA. TRIAL REGISTRATION NUMBER: ChiCTR1900022488.

Prevotella copri promotes vascular calcification via lipopolysaccharide through activation of NF-κB signaling pathway.

Emerging evidence indicates that alteration of gut microbiota plays an important role in chronic kidney disease (CKD)-related vascular calcification (VC). We aimed to investigate the specific gut microbiota and the underlying mechanism involved in CKD-VC. We identified an increased abundance of Prevotella copri (P. copri) in the feces of CKD rats (induced by using 5/6 nephrectomy followed by a high calcium and phosphate diet) with aortic calcification via amplicon sequencing of 16S rRNA genes. In patients with CKD, we further confirmed a positive correlation between abundance of P. copri and aortic calcification scores. Moreover, oral administration of live P. copri aggravated CKD-related VC and osteogenic differentiation of vascular smooth muscle cells in vivo, accompanied by intestinal destruction, enhanced expression of Toll-like receptor-4 (TLR4), and elevated lipopolysaccharide (LPS) levels. In vitro and ex vivo experiments consistently demonstrated that P. copri-derived LPS (Pc-LPS) accelerated high phosphate-induced VC and VSMC osteogenic differentiation. Mechanistically, Pc-LPS bound to TLR4, then activated the nuclear factor κB (NF-κB) and nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) inflammasome signals during VC. Inhibition of NF-κB reduced NLRP3 inflammasome and attenuated Pc-LPS-induced VSMC calcification. Our study clarifies a novel role of P. copri in CKD-related VC, by the mechanisms involving increased inflammation-regulating metabolites including Pc-LPS, and activation of the NF-κB/NLRP3 signaling pathway. These findings highlight P. copri and its-derived LPS as potential therapeutic targets for VC in CKD.

Remnant cholesterol and the risk of aortic valve calcium progression: insights from the MESA study.

BACKGROUND: Remnant cholesterol (RC) is implicated in the risk of cardiovascular disease. However, comprehensive population-based studies elucidating its association with aortic valve calcium (AVC) progression are limited, rendering its precise role in AVC ambiguous. METHODS: From the Multi-Ethnic Study of Atherosclerosis database, we included 5597 individuals (61.8 ± 10.1 years and 47.5% men) without atherosclerotic cardiovascular disease at baseline for analysis. RC was calculated as total cholesterol minus high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C), as estimated by the Martin/Hopkins equation. Using the adjusted Cox regression analyses, we examined the relationships between RC levels and AVC progression. Furthermore, we conducted discordance analyses to evaluate the relative AVC risk in RC versus LDL-C discordant/concordant groups. RESULTS: During a median follow-up of 2.4 ± 0.9 years, 568 (10.1%) participants exhibited AVC progression. After adjusting for traditional cardiovascular risk factors, the HRs (95% CIs) for AVC progression comparing the second, third, and fourth quartiles of RC levels with the first quartile were 1.195 (0.925-1.545), 1.322 (1.028-1.701) and 1.546 (1.188-2.012), respectively. Notably, the discordant high RC/low LDL-C group demonstrated a significantly elevated risk of AVC progression compared to the concordant low RC/LDL-C group based on their medians (HR, 1.528 [95% CI 1.201-1.943]). This pattern persisted when clinical LDL-C threshold was set at 100 and 130 mg/dL. The association was consistently observed across various sensitivity analyses. CONCLUSIONS: In atherosclerotic cardiovascular disease-free individuals, elevated RC is identified as a residual risk for AVC progression, independent of traditional cardiovascular risk factors. The causal relationship of RC to AVC and the potential for targeted RC reduction in primary prevention require deeper exploration.

Cardiomyocyte Atrophy, an Underestimated Contributor in Doxorubicin-Induced Cardiotoxicity.

Left ventricular (LV) mass loss is prevalent in doxorubicin (DOX)-induced cardiotoxicity and is responsible for the progressive decline of cardiac function. Comparing with the well-studied role of cell death, the part of cardiomyocyte atrophy (CMA) playing in the LV mass loss is underestimated and the knowledge of the underlying mechanism is still limited. In this review, we summarized the recent advances in the DOX-induced CMA. We found that the CMA caused by DOX is associated with the upregulation of FOXOs and "atrogenes," the activation of transient receptor potential canonical 3-NADPH oxidase 2 (TRPC3-Nox2) axis, and the suppression of IGF-1-PI3K signaling pathway. The imbalance of anabolic and catabolic process may be the common final pathway of these mechanisms. At last, we provided some strategies that have been demonstrated to alleviate the DOX-induced CMA in animal models.

Spermidine-enhanced autophagic flux improves cardiac dysfunction following myocardial infarction by targeting the AMPK/mTOR signalling pathway.

BACKGROUND AND PURPOSE: Spermidine, a natural polyamine, is abundant in mammalian cells and is involved in cell growth, proliferation, and regeneration. Recently, oral spermidine supplements were cardioprotective in age-related cardiac dysfunction, through enhancing autophagic flux. However, the effect of spermidine on myocardial injury and cardiac dysfunction following myocardial infarction (MI) remains unknown. EXPERIMENTAL APPROACH: We determined the effects of spermidine in a model of MI, Sprague-Dawley rats with permanent ligation of the left anterior descending artery, and in cultured neonatal rat cardiomyocytes (NRCs) exposed to angiotensin II (Ang II). Cardiac function in vivo was assessed with echocardiography. In vivo and in vitro studies used histological and immunohistochemical techniques, along with western blots. KEY RESULTS: Spermidine improved cardiomyocyte viability and decreased cell necrosis in NRCs treated with angiotensin II. In rats post-MI, spermidine reduced infarct size, improved cardiac function, and attenuated myocardial hypertrophy. Spermidine also suppressed the oxidative damage and inflammatory cytokines induced by MI. Moreover, spermidine enhanced autophagic flux and decreased apoptosis both in vitro and in vivo. The protective effects of spermidine on cardiomyocyte apoptosis and cardiac dysfunction were abolished by the autophagy inhibitor chloroquine, indicating that spermidine exerted cardioprotective effects at least partly through promoting autophagic flux, by activating the AMPK/mTOR signalling pathway. CONCLUSIONS AND IMPLICATIONS: Our findings suggest that spermidine improved MI-induced cardiac dysfunction by promoting AMPK/mTOR-mediated autophagic flux.

Sulforaphane prevents rat cardiomyocytes from hypoxia/reoxygenation injury in vitro via activating SIRT1 and subsequently inhibiting ER stress.

AIM: Sulforaphane (SFN), a natural dietary isothiocyanate, is found to exert beneficial effects for cardiovascular diseases. This study aimed to investigate the mechanisms underlying the protective effects of SFN in a model of myocardial hypoxia/reoxygenation (H/R) injury in vitro. METHODS: Cultured neonatal rat cardiomyocytes pretreated with SFN were subjected to 3-h hypoxia followed by 3-h reoxygenation. Cell viability and apoptosis were detected. Caspase-3 activity and mitochondrial membrane potential (ΔΨm) was measured. The expression of ER stress-related apoptotic proteins were analyzed with Western blot analyses. Silent information regulator 1 (SIRT1) activity was determined with SIRT1 deacetylase fluorometric assay kit. RESULTS: SFN (0.1-5 µmol/L) dose-dependently improved the viability of cardiomyocytes, diminished apoptotic cells and suppressed caspase-3 activity. Meanwhile, SFN significantly alleviated the damage of ΔΨm and decreased the expression of ER stress-related apoptosis proteins (GRP78, CHOP and caspase-12), elevating the expression of SIRT1 and Bcl-2/Bax ratio in the cardiomyocytes. Co-treatment of the cardiomyocytes with the SIRT1-specific inhibitor Ex-527 (1 µmol/L) blocked the SFN-induced cardioprotective effects. CONCLUSION: SFN prevents cardiomyocytes from H/R injury in vitro most likely via activating SIRT1 pathway and subsequently inhibiting the ER stress-dependent apoptosis.

[Changes in serum level of carboxy-terminal telopeptide of type I collagen in patients with coronary heart disease].

OBJECTIVE: To investigate the serum level of carboxy-terminal telopeptide of type I collagen (ICTP) and explore its correlation with MMP-2 and MMP-9 in patients with coronary artery disease (CHD). METHODS: A total of 103 CHD patients treated in our hospital between October, 2013 and May, 2014 were enrolled, including 39 with stable angina pectoris (SAP), 39 with unstable angina (UA), and 25 with acute myocardial infarction (AMI), with 38 non-CHD volunteers as the control group. The serum levels of ICTP, MMP-2, and MMP-9 were detected in all the subjects using enzyme-linked immunosorbent assay (ELISA). RESULTS: No significant difference in serum levels of MMP-2, MMP-9, or ICTP was found between the control and SAP groups or between UA and AMI groups (P>0.05), but the latter two groups had significantly higher serum levels of MMP-2, MMP-9, and ICTP than the former two groups (P<0.05). Serum ICTP level was found to negatively correlated with the fibrotic area and positively with the lipid component in the plaques (P<0.05). Regression analysis revealed significant positive correlations of serum ICTP with MMP-2 and MMP-9 (P<0.05). CONCLUSION: An elevated serum ICTP level is indicative of the presence of unstable plaques in CHD patients. Serum ICTP is more strongly correlated with MMP-2 than with MMP-9, and can be used as a non-invasive marker for assessing vulnerable plaques in patients with acute coronary syndrome.

Trans-subclavian approach for radiofrequency ablation of premature ventricular contractions originating from subtricuspid annulus: a case report.

BACKGROUND: Catheter ablation has been established as a curative treatment strategy for ventricular arrhythmias. The standard procedure of most ventricular arrhythmias originating from the right ventricle is performed via the femoral vein. However, a femoral vein access may not achieve a successful ablation in some patients. CASE PRESENTATION: We reported a case of a 29-year old patient with symptomatic premature ventricular contractions was referred for catheter ablation. Radiofrequency energy application at the earliest endocardial ventricular activation site via the right femoral vein could not eliminate the premature ventricular contractions. Epicardial mapping could not obtain an earlier ventricular activation when compared to the endocardial mapping, and at the earliest epicardial site could not provide an identical pace mapping. Finally, we redeployed the ablation catheter via the right subclavian vein by a long sheath. During mapping of the subvalvular area of the right ventricle, a site with a good pace mapping and early ventricular activation was found, and premature ventricular contractions were eliminated successfully. CONCLUSION: Ventricular arrhythmias originating from the subtricuspid annulus may be successfully abolished via a trans-subclavian approach and a long sheath. Although access via the right subclavian vein for mapping and ablation is an effective alternative, it is not a routine approach.

[Predictors of occurrence of left atrium and left appendage thrombosis in patients with nonvalvular atrial fibrillation].

OBJECTIVE: To determine the predictors of thrombosis in left atrium (LA) or left atrial appendage (LAA) in patients with nonvalvular atrial fibrillation. METHODS: Two hundred and eight patients under 65 year old with atrial fibrillation (AF) were included and all of them received examination of transesophageal echocardiography (TEE). Thrombus formation in LA/LAA was found in 23 patients (thrombus group) but absent in the remaining 185 patients (nonthrombus group). All patients were analyzed by univariate regression and binary logistic regression to investigate the relationship between the occurrence of LA/LAA thrombosis and these factors (such as case history, smoking/drinking preference, indicators of clinical blood examination and ultrasound imaging study, etc) RESULTS: Univariate analysis revealed that diameter of LA [(34.9 +/- 4.4) mm vs (42.2 +/- 6.5) mm, P = 0.000], ratio of chest and heart (60/185 vs 20/23 P = 0.000), brain infarction/transient ischemic attack (TAI) (7/185 vs 6/23 P = 0.000), smoking (30/185 vs 8/23, P = 0.030), fibrinogen (FIB) [(3.0 +/- 0.7)g/L vs (3.5 +/- 1.0) g/L, P = 0.000], coronary artery disease (CAD) (10/185 vs 6/23, P = 0.000) and LVDd [(45.7 +/- 4.1) mm vs (48.5 +/- 5.7) mm, P = 0.000] and LVEF [(65.1 +/- 6.6) mm vs (59.3 +/- 1.3) mm, P = 0.050] were significant between nonthrombus group and thrombus group (P < 0.05). However binary logistic regression analysis identified that only LAD, ratio of chest and heart, brain infarction/TAI and FIB were the significant and independent predictors of LA/LAA thrombosis. CONCLUSION: Diameter of LA, ratio of chest and heart, brain infarction/TAI and FIB are independent risk factors of thrombosis in patients under 65 year old with nonvalvular atrial fibrillation. These patients need a better anticoagulation.

[Left atrial size and function after radiofrequency catheter ablation for paroxysmal atrial fibrillation].

OBJECTIVE: To evaluate the impact of radiofrequency catheter ablation on left atrial (LA) size and function in patients with paroxysmal atrial fibrillation (PAF) and whether there is any difference between segmental pulmonary vein ostial isolation (SPVI) and circumferential pulmonary vein ablation (CPVA). METHODS: Sixty-six patients with highly symptomatic atrial fibrillation were assigned to undergo either SPVI or CPVA. Transthorax echocardiography was performed before, 1 day, 1 months and 3 months after the procedure. LA dimension, LA area, late diastolic peak velocity of mitral valve inflow (A) and peak atrial systolic mitral annulus velocity (A') were recorded. RESULTS: Of 66 consecutive patients with symptomatic PAF, 30 patients underwent SPVI and 36 underwent CPVA. After a mean follow-up of (315 +/- 153) days, 21 patients (70%) after SPVI and 28 patients (75%) after CPVA were free of atrial tachyarrhythmia. As compared with the baseline, LA area decreased at 1-month after ablation in SPVI group and at 3-month in CPVA group. LA dimension decreased also in SPVI group, but did not in CPVA group. A velocity and A' velocity declined remarkably 1 day after CPVA, and restored 3 months later. The former went back to the level of baseline, and the latter exceeded it apparently. In SPVI group, A velocity increased at 1-month, and maintained in 3-month after ablation. A' velocity increased at 3-month after ablation. No reduction of A velocity or A' velocity was found after SPVI. CONCLUSIONS: This study demonstrated a decrease in LA area and an improvement in LA systolic function 3 months after ablation for PAF. The LA damage by CPVA was more than that by SPVI, which was characterized by the reduction of LA function 1 day after procedure and the delayed improvement of LA size and functional parameters.

[Radiofrequency ablation of idiopathic right ventricular tachycardia].

OBJECTIVE: To study the effect of electrophysiological characterization and radiofrequency ablation on idiopathic right ventricular tachycardia. METHODS: Five patients ( 3 male and 2 female ) with an average age of 35.2+/-11.2 years were enrolled in this study. 7F EPT electrode in temperature-controlled mode was used for the ablation. The temperature and power were controlled within the range of 50-55 degrees Celsius; and 30-35 W respectively. The ablation target was the point that evidently induced P-potential at posterior right ventricular septum by scaling test. Tachycardia was stopped within 3 s. Consolidated discharge was within 40 s. The ablation was ended when the tachycardia could not be evoked during routine intracardiac electrophysiology test. Aspirin (0.1 g/d) was given orally for 1 month after operation. RESULTS: The body surface electrocardiogram did not change significantly after ablation. Neither S1S1 and program med stimulation, nor intravenous drip of isoproterenol after operation evoked tachycardia. Neither tachycardia nor complication appeared 4-6 months after the test. CONCLUSIONS: (1) Posterior right interventricular septum can also give rise to idiopathic ventricular tachycardia similar to left posterior interventricular septum. Ablation at the point where P-potential can be evidently induced by scaling test could easily acquire success. (2) Idiopathic right ventricular tachycardia has typical body surface electrocardiogram when tachycardia attacks. (3) Ventricular tachycardia is different from bundle branch reciprocal ventricular tachycardia.

[Effects of C-type natriuretic peptide gene transduction on neointimal hyperplasia and endothelial function after angioplasty].

OBJECTIVE: To investigate the effects of C-type natriuretic peptide (CNP) gene transduction on neointimal hyperplasia and endothelial function after angioplasty. METHODS: Eighty-four rabbits were divided into 3 equal groups, namely normal control group, alkaline phosphatase gene transduction group and CNP gene transduction group. The rabbits in the latter two groups were given high-cholesterol diet 7 d before the experiment, followed by establishment of restenosis models by injuring the iliac artery and the specified gene transfer via retroviral vectors. Those in the normal control group were fed with normal diet. Before high-cholesterol diet and killing respectively, 2 ml venous blood samples were taken for testing blood lipid and serum CNP concentration. In the two groups with gene transduction, the injured rabbit iliac arteries were harvested for ex vivo vascular ring tension test, histological and pathological examinations, as well as immunohistochemistry analysis of CNP. The lumen area, neointimal thickness, neointimal area, ratio of intimal to medial area were measured by image analysis system. RESULTS: There were no significant differences in blood lipid and serum CNP concentration between the two gene transduction groups at the same time points both before and after operation. In CNP gene transduction group, endothelium-dependent relaxation of the vascular rings was significantly improved in comparison with the other two groups (P<0.01), irrespective of L-Arg pretreatment, whereas endothelium-independent relaxation function varied little between the 3 groups (P>0.05). Poor relaxation function to Ach of the vascular rings was resulted after pretreatment with LMMA. CNP gene expression at the site of gene transfer was detected in the CNP gene transduction group and in 2 weeks after balloon injury, the neointimal thickness, neointimal area and ratio of the neointimal to tunica media area were markedly increased in the two gene transduction groups, but the measurements were significantly lower in CNP group (P<0.01). CONCLUSION: CNP gene can be successfully transferred and effectively expressed at the injured site in the blood vessels to decrease the hyperplasia and significantly improve endothelial function after angioplasty.

[Ablation of postoperative "incisional" reentrant atrial tachycardia and flutter in children using the CARTO system].

OBJECTIVE: In children with congenital heart diseases who have undergone surgical interventions, postoperative arrhythmias frequently complicate the clinical course. "Incisional" atrial tachycardia or flutter is one of the most common forms of postoperative arrhythmias in these patients and can lead to significant morbidity and even mortality. The aim of this study was to investigate how to use antiarrhythmic drugs and the CARTO system to treat these cases. METHODS: There were 12 patients with "incisional" atrial tachycardia or flutter complicating surgery for congenital heart diseases in this study (3 patients with correction of tetrology of Fallot, 3 with atrial septal defect repair, 2 with ventricular septal defect repair, 1 with switch, 1 with repair of Ebstein's anomaly, 1 with total anomalous pulmonary venous drainage, and 1 with atrial septal closure with the Amplatzer septal occlusion). Patients whose body weight was less than 10 kg or those who did not wish to accept ablation were treated with antiarrhythmic drugs, including digitoxin, propranolol, metoprolol and cordarone. CARTO system was used to map 6 patients whose body weight was more than 10 kg and who agreed with accepting ablation for atrial tachycardia and flutter. Radio-frequency ablation was performed in these 6 cases including two cases of "incisional" atrial tachycardia and 4 of atrial flutter. RESULTS: (1) The antiarrhythmic drug was successful in 6 patients with "incisional" atrial tachycardia. (2) Six patients including 2 children with "incisional" atrial tachycardia and 4 children with atrial flutter were successfully ablated. But one case of "incisional" atrial tachycardia relapsed after 3 months of ablation. This case, however, was successfully ablated again later. No further relapse was observed during the 2 - 24 months of follow-up. CONCLUSION: Ablation of "incisional" atrial tachycardia and flutter is the first choice to treat the patients whose body weight is more than 10 kg and those who agree with accepting ablation by CARTO system. Drug therapy of "incisional" atrial tachycardia and flutter is palliative and it is the only selection to treat the patients whose body weight is less than 10 kg or those who do not wish to accept ablation procedure.