Beneficial Cardiovascular Effects Research Articles

NaHS restores the vascular alterations in the renin-angiotensin system induced by hyperglycemia in rats

Hyperglycemia (HG) impairs the renin-angiotensin system (RAS), which may contribute to vascular dysfunction. Besides, hydrogen sulfide (H2S) exerts beneficial cardiovascular effects in metabolic diseases. Therefore, our study aimed to determine the effects of chronic administration of sodium hydrosulfide (NaHS; inorganic H2S donor) and DL-Propargylglycine [DL-PAG; cystathionine-ץ-lyase (CSE) inhibitor] on the RAS-mediated vascular responses impairments observed in thoracic aortas from male diabetic Wistar rats. For that purpose, neonatal rats were divided into two groups that received: 1) citrate buffer (n = 12) or 2) streptozotocin (STZ, 70 mg/kg; n = 48) on the third postnatal day. After 12 weeks, diabetic animals were divided into 4 subgroups (n = 12 each) that received daily i.p. injections during 4 weeks of: 1) non-treatment; 2) vehicle (PBS, 1 mL/kg); 3) NaHS (5.6 mg/kg); and 4) DL-PAG (10 mg/kg). After treatments (16 weeks), blood glucose, angiotensin-(1−7) [Ang-(1−7)], and angiotensin II (Ang II) levels, vascular responses to Ang-(1−7) and Ang II, and the expression of angiotensin AT1, AT2, and Mas receptors, angiotensin converting enzyme (ACE) and ACE type 2 (ACE2) were determined. HG induced: 1) increased blood glucose levels and expression of angiotensin II AT1 receptor; 2) impaired Ang-(1−7) and Ang II mediated vascular responses; 3) decreased angiotensin levels and expression of angiotensin II AT2 and angiotensin-(1−7) Mas receptors, and ACE2; and 4) no changes in ACE expression. Interestingly, NaHS, but not DL-PAG, reversed HG-induced impairments, except for blood glucose level changes. These results suggest that NaHS restores vascular function in streptozotocin-induced HG through RAS modulation.

Sodium-glucose cotransporter-2 inhibitors and cardiovascular clinical outcomes in acute heart failure: A narrative review.

In an effort to expedite the publication of articles, AJHP is posting manuscripts online as soon as possible after acceptance. Accepted manuscripts have been peer-reviewed and copyedited, but are posted online before technical formatting and author proofing. These manuscripts are not the final version of record and will be replaced with the final article (formatted per AJHP style and proofed by the authors) at a later time. This review describes the evidence from randomized controlled trials (RCTs) regarding the effects of sodium-glucose cotransporter 2 (SGLT2) inhibitors on cardiovascular (CV) clinical outcomes when therapy is initiated during acute heart failure (HF). SGLT2 inhibitors have become a cornerstone of guideline-directed medical therapy (GDMT) for type 2 diabetes mellitus, chronic kidney disease, and HF. Because of their ability to promote natriuresis and diuresis as well as other potentially beneficial CV effects, use of SGLT2 inhibitors has been investigated when therapy is initiated during hospitalization for acute HF. We identified 5 placebo-controlled RCTs that reported CV clinical outcomes incorporating one or more components of all-cause mortality, CV mortality, CV hospitalization, HF worsening, and hospitalization for HF in patients treated with empagliflozin (n = 3 trials), dapagliflozin (n = 1 trial), and sotagliflozin (n = 1 trial). Nearly all CV outcomes in these trials showed benefit with SGLT2 inhibitor use during acute HF. Incidence of hypotension, hypokalemia, and acute renal failure was generally similar to that with placebo. These findings are limited by heterogeneous outcome definitions, variation in time to SGLT2 inhibitor initiation, and small sample sizes. SGLT2 inhibitors may have a role in inpatient management of acute HF, provided there is close monitoring for fluctuations in hemodynamic, fluid, and electrolyte status. Initiation of SGLT2 inhibitors at the time of acute HF may promote optimized GDMT, continued medication adherence, and reduced risk of CV outcomes.

Human epicardial adipose tissue expresses glucose-dependent insulinotropic polypeptide, glucagon, and glucagon-like peptide-1 receptors as potential targets of pleiotropic therapies.

Human epicardial adipose tissue (EAT) plays a crucial role in the development and progression of coronary artery disease, atrial fibrillation, and heart failure. Microscopically, EAT is composed of adipocytes, nerve tissues, inflammatory, stromovascular, and immune cells. Epicardial adipose tissue is a white adipose tissue, albeit it also has brown fat-like or beige fat-like features. No muscle fascia divides EAT and myocardium; this allows a direct interaction and crosstalk between the epicardial fat and the myocardium. Thus, it might be a therapeutic target for pharmaceutical compounds acting on G-protein-coupled receptors, such as those for glucose-dependent insulinotropic polypeptide (GIP), glucagon (GCG), and glucagon-like peptide-1 (GLP-1), whose selective stimulation with innovative drugs has demonstrated beneficial cardiovascular effects. The precise mechanism of these novel drugs and their tissue and cellular target(s) need to be better understood. We evaluate whether human EAT expresses GIP, GCG, and GLP-1 receptors and whether their presence is related to EAT transcriptome. We also investigated protein expression and cell-type localization specifically for GIP receptor (GIPR) and glucagon receptor (GCGR). Epicardial adipose tissue samples were collected from 33 patients affected by cardiovascular diseases undergoing open heart surgery (90.9% males, age 67.2 ± 10.5 years mean ± SD). Microarray and immunohistochemistry analyses were performed. Microarray analysis showed that GIPR and GCGR messenger ribonucleic acids (mRNAs) are expressed in EAT, beyond confirming the previously found GLP-1 [3776 ± 1377 arbitrary unit (A.U.), 17.77 ± 14.91 A.U., and 3.41 ± 2.27 A.U., respectively]. The immunohistochemical analysis consistently indicates that GIPR and GCGR are expressed in EAT, mainly in macrophages, isolated, and in crown-like structures. In contrast, only some mature adipocytes of different sizes showed cytoplasmic immunostaining, similar to endothelial cells and pericytes in the capillaries and pre-capillary vascular structures. Notably, EAT GIPR is statistically associated with the low expression of genes involved in free fatty acid (FFA) oxidation and transport and those promoting FFA biosynthesis and adipogenesis (P < 0.01). Epicardial adipose tissue GCGR, in turn, is related to genes involved in FFA transport, mitochondrial fatty acid oxidation, and white-to-brown adipocyte differentiation, in addition to genes involved in the reduction of fatty acid biosynthesis and adipogenesis (P < 0.01). Having reported the expression of the GLP-1 receptor previously, here, we showed that GIPR and GCGR similarly present at mRNA and protein levels in human EAT, particularly in macrophages and partially adipocytes, suggesting these G-protein-coupled receptors as pharmacological targets on the ongoing innovative drugs, which seem cardiometabolically healthy well beyond their effects on glucose and body weight.

Effects of empagliflozin on left ventricular diastolic function in addition to usual care in individuals with type 2 diabetes mellitus—results from the randomized, double-blind, placebo-controlled EmDia trial

BackgroundThe sodium-glucose co-transporter 2 inhibitor empagliflozin improves cardiovascular outcome in patients with type 2 diabetes mellitus (T2DM) and heart failure. Experimental studies suggest a direct cardiac effect of empagliflozin associated with an improvement in left ventricular diastolic function.MethodsIn the randomized, double-blind, two-armed, placebo-controlled, parallel group trial EmDia, patients with T2DM and elevated left ventricular E/E´ ratio were enrolled and randomized 1:1 to receive empagliflozin 10 mg/day versus placebo. The primary endpoint was the change of left ventricular E/E´ ratio after 12 weeks of intervention.ResultsA total of 144 patients with T2DM and an elevated left ventricular E/e´ ratio (age 68.9 ± 7.7 years; 14.1% women; E/e´ ratio 9.61[8.24/11.14], left ventricular ejection fraction 58.9% ± 5.6%). After 12 weeks of intervention, empagliflozin resulted in a significant higher decrease in the primary endpoint E/e´ ratio by − 1.18 ([95% confidence interval (CI) − 1.72/− 0.65]; P < 0.0001) compared with placebo. The beneficial effect of empagliflozin was consistent across all subgroups and also occurred in subjects with heart failure and preserved ejection fraction (n = 30). Additional effects of empagliflozin on body weight, HbA1c, uric acid, red blood cell count, hemoglobin, mean corpuscular hemoglobin, and hematocrit were detected (all P < 0.001). Approximately one-third of the reduction in E/e´ by empagliflozin could be explained by the variables examined.ConclusionsEmpagliflozin improves diastolic function in patients with T2DM and elevated end-diastolic pressure. Since the positive effects were consistent in patients with and without heart failure with preserved ejection fraction, the data add a mechanistic insight for the beneficial cardiovascular effect of empagliflozin.Trial registrationClinicaltrials.gov, unique identifier: NCT02932436.Graphical abstract

The Sodium-Glucose Co-Transporter 2 (SGLT2) Inhibitor Empagliflozin Reverses Hyperglycemia-Induced Monocyte and Endothelial Dysfunction Primarily through Glucose Transport-Independent but Redox-Dependent Mechanisms

Hyperglycaemia-induced oxidative stress and inflammation contribute to vascular cell dysfunction and subsequent cardiovascular events in T2DM. Selective sodium-glucose co-transporter-2 (SGLT-2) inhibitor empagliflozin significantly improves cardiovascular mortality in T2DM patients (EMPA-REG trial). Since SGLT-2 is known to be expressed on cells other than the kidney cells, we investigated the potential ability of empagliflozin to regulate glucose transport and alleviate hyperglycaemia-induced dysfunction of these cells. Primary human monocytes were isolated from the peripheral blood of T2DM patients and healthy individuals. Primary human umbilical vein endothelial cells (HUVECs) and primary human coronary artery endothelial cells (HCAECs), and fetoplacental endothelial cells (HPECs) were used as the EC model cells. Cells were exposed to hyperglycaemic conditions in vitro in 40 ng/mL or 100 ng/mL empagliflozin. The expression levels of the relevant molecules were analysed by RT-qPCR and confirmed by FACS. Glucose uptake assays were carried out with a fluorescent derivative of glucose, 2-NBDG. Reactive oxygen species (ROS) accumulation was measured using the H2DFFDA method. Monocyte and endothelial cell chemotaxis were measured using modified Boyden chamber assays. Both primary human monocytes and endothelial cells express SGLT-2. Hyperglycaemic conditions did not significantly alter the SGLT-2 levels in monocytes and ECs in vitro or in T2DM conditions. Glucose uptake assays carried out in the presence of GLUT inhibitors revealed that SGLT-2 inhibition very mildly, but not significantly, suppressed glucose uptake by monocytes and endothelial cells. However, we detected the significant suppression of hyperglycaemia-induced ROS accumulation in monocytes and ECs when empagliflozin was used to inhibit SGLT-2 function. Hyperglycaemic monocytes and endothelial cells readily exhibited impaired chemotaxis behaviour. The co-treatment with empagliflozin reversed the PlGF-1 resistance phenotype of hyperglycaemic monocytes. Similarly, the blunted VEGF-A responses of hyperglycaemic ECs were also restored by empagliflozin, which could be attributed to the restoration of the VEGFR-2 receptor levels on the EC surface. The induction of oxidative stress completely recapitulated most of the aberrant phenotypes exhibited by hyperglycaemic monocytes and endothelial cells, and a general antioxidant N-acetyl-L-cysteine (NAC) was able to mimic the effects of empagliflozin. This study provides data indicating the beneficial role of empagliflozin in reversing hyperglycaemia-induced vascular cell dysfunction. Even though both monocytes and endothelial cells express functional SGLT-2, SGLT-2 is not the primary glucose transporter in these cells. Therefore, it seems likely that empagliflozin does not directly prevent hyperglycaemia-mediated enhanced glucotoxicity in these cells by inhibiting glucose uptake. We identified the reduction of oxidative stress by empagliflozin as a primary reason for the improved function of monocytes and endothelial cells in hyperglycaemic conditions. In conclusion, empagliflozin reverses vascular cell dysfunction independent of glucose transport but could partially contribute to its beneficial cardiovascular effects.

ELITE: rationale and design of a longitudinal elite athlete, extreme cardiovascular phenotyping, prospective cohort study

IntroductionThe cardiovascular benefits of physical exercise are well-known. However, vigorous exercise has also been associated with adverse cardiac effects. To improve our understanding of cardiovascular adaptation to exercise versus maladaptation...

Flavan-3-ol-methylxanthine interactions: Modulation of flavan-3-ol bioavailability in volunteers with a functional colon and an ileostomy.

Flavan-3-ols, including the flavan-3-ol monomer (-)-epicatechin, are dietary bioactives known to mediate beneficial cardiovascular effects in humans. Recent studies showed that flavan-3-ols could interact with methylxanthines, evidenced by an increase in flavan-3-ol bioavailability with a concomitant increase in flavan-3-ol intake-mediated vascular effects. This study aimed at elucidating flavan-3-ol-methylxanthine interactions in humans in vivo by evaluating the specific contributions of theobromine and caffeine on flavan-3-ol bioavailability. In ileostomists, the effect of methylxanthines on the efflux of flavan-3-ol metabolites in the small intestine was assessed, a parameter important to an understanding of the pharmacokinetics of flavan-3-ols in humans. In a randomized, controlled, triple cross-over study in volunteers with a functional colon (n=10), co-ingestion of flavan-3-ols and cocoa methylxanthines, mainly represented by theobromine, increased peak circulatory levels (Cmax) of flavan-3-ols metabolites (+21±8%; p<0.05). Conversely, caffeine did not mediate a statistically significant effect on flavan-3-ol bioavailability (Cmax=+10±8%, p=n.s.). In a subsequent randomized, controlled, double cross-over study in ileostomists (n=10), cocoa methylxanthines did not affect circulatory levels of flavan-3-ol metabolites, suggesting potential differences in flavan-3-ol bioavailability compared to volunteers with a functional colon. The main metabolite in ileal fluid was (-)-epicatechin-3'-sulfate, however, no differences in flavan-3-ol metabolites in ileal fluid were observed after flavan-3-ol intake with and without cocoa methylxanthines. Taken together, these results demonstrate a differential effect of caffeine and theobromine in modulating flavan-3-ol bioavailability when these bioactives are co-ingested. These findings should be considered when comparing the effects mediated by the intake of flavan-3-ol-containing foods and beverages and the amount and type of methylxanthines present in the ingested matrixes. Ultimately, these insights will be of value to further optimize current dietary recommendations for flavan-3-ol intake. CLINICAL TRIAL REGISTRATION NUMBER: This work was registered at clinicaltrials.gov as NCT03526107 (study part 1, volunteers with functional colon) and NCT03765606 (study part 2, volunteers with an ileostomy).

News in diabetology: what's new in 2022

Diabetology is a constantly evolving discipline, many molecules appear on the market and treatment recommendations change quite frequently according to the latest published studies. It is therefore often difficult for the primary care physician to be up to date. After lifestyle measures that must of course be preferred before any drug approach, metformin remains the pharmacological basis of treatment. Current recommendations favor the introduction of an SGLT2 inhibitor or a GLP-1 receptor agonist after metformin because these molecules have shown beneficial cardiovascular and renal effects. The purpose of this article is to help the primary care physician to choose the most suitable pharmacological treatment for each patient, in the light of the 2022 novelties in the field of diabetes.

Role of cytochrome P450-epoxygenase and soluble epoxide hydrolase in the regulation of vascular response.

The role of cytochrome P450-epoxygenase has been seen in cardiovascular physiology and pathophysiology. The aberration in CYP450-epoxygenase genes occur due to genetic polymorphisms, aging, or environmental factors, that increase susceptibility to cardiovascular diseases (CVDs). The actual role played by the CYP450-epoxygenases is the metabolism of arachidonic acid (AA) and linoleic acid (LA) into epoxyeicosatrienoic acids (EETs) and epoxyoctadecaenoic acid (EpOMEs) metabolites (oxylipins) and others, which is involved in vasodilation and myocardial-protection. But the genetic polymorphisms in CYP450-epoxygenases lose their beneficial cardiovascular effects of oxylipins, and the soluble epoxide hydrolase (sEH) antagonizes beneficial oxylipins into diols. Like sEH converts EETs into dihydroxyeicosatrienoic acid (DHETs), EpOMEs into dihydroxyoctadecaenoic acid (DiHOMEs), and reverses its beneficial effects, and the sEH gene (Ephx2) polymorphisms cause the enzyme to become overactive and convert epoxy-fatty acids into diols, making them vulnerable to CVDs, including hypertension. Other, enzymes like ω-hydroxylases (CYP450-4A11 & CYP450-4F2)-derived oxylipins from AA, ω-terminal-hydroxyeicosatetraenoic acids (19-, 20-HETE), lipoxygenase-derived oxylipins, mid-chain hydroxyeicosatetraenoic acids (5-, 11-, 12-, 15-HETEs), and the cyclooxygenase-derived prostanoids (prostaglandins: PGD2, PGF2α; thromboxane: Txs, oxylipins) are involved in vasoconstriction, hypertension, inflammation, and cardiac toxicity. Also, there are significant interactions were seen between adenosine receptors [adenosine A2A receptor (A2AAR) and adenosine A1 receptor (A1AR)] with CYP450-epoxygenases, ω-hydroxylases, sEH, and their derived oxylipins in the regulation of the cardiovascular response. Moreover, polymorphisms exist in CYP450-epoxygenases, ω-hydroxylases, sEH, and the adenosine receptor genes in populations associated with CVDs. This chapter will discuss the role of oxylipins' interactions with adenosine receptors in cardiovascular function/dysfunction in mice and humans.

Camellia oil (Camellia oleifera Abel.) treatment improves high-fat diet-induced atherosclerosis in apolipoprotein E (ApoE)-/- mice.

Atherosclerosis is the main cause of cardiovascular diseases, and healthy dietary habits are a feasible strategy to prevent atherosclerosis development. Camellia oil, an edible plant oil, exhibits multiple beneficial cardiovascular effects. Our previous study showed that oral administration of camellia oil attenuated hyperglycemia, fat deposits in the liver, and the atherosclerosis index in high-fat diet (HFD)-induced obese mice. Here, an atherosclerosis model of apolipoprotein E (ApoE)-/- mice induced by HFD was used to study the effect of camellia oil on atherosclerosis, and 16S rRNA gene sequencing was used to analyze the changes in gut microbiota composition. The results showed that camellia oil significantly inhibited the formation of atherosclerotic plaques in ApoE-/- mice, which were characterized by significantly reduced levels of serum total cholesterol and enhanced levels of serum high-density lipoprotein cholesterol. The aortic levels of interleukin-6 and tumor necrosis factor were decreased. The results of the 16S rRNA analysis showed that after camellia oil interventions, the intestinal flora of ApoE-/- mice changed significantly, with the diversity of intestinal flora especially increasing, the relative abundances of Bacteroides, Faecalibaculum, Bilophila, and Leuconostoc increasing, and the Firmicutes/Bacteroidetes ratio and Firmicutes abundance decreasing. Collectively, our findings confirmed the promising value of camellia oil in preventing the development of atherosclerosis in ApoE-/- mice. Mechanistically, this preventive effect of camellia oil was probably due to its lipid-lowering activity, anti-inflammatory effects, and alteration of the gut microbiota composition in the mice.

Sodium Glucose Co-Transporter-2 (SGLT 2) Inhibitor as A Therapy For Heart Failure With Low Ejection Fraction (HFrEF)

Heart failure is a major cause of cardiovascular morbidity and mortality worldwide despite advances in prevention and management. Recent clinical trial findings have shown that sodium-glucose co-transporter 2 inhibitor (SGLT2i) produces effects other than lowering blood glucose levels, i.e., exhibits beneficial cardiovascular effects. The purpose of this paper is to determine the role of SGLT2i in heart failure patients with low ejection fraction (HFrEF) so that it can be used as a guide for future management. This study is a narrative review by searching on Google Scholar through several keywords. The period including articles obtained is articles from 2014 to 2022. The diuretic effect of SGLT2i can cause an increase in cardiac preload. The reduction in cardiac afterload likely occurs through a decrease in blood pressure and arterial stiffness resulting in increased endocardial blood flow. SGLT2i has a beneficial effect on the heart remodeling process. SGLT2i can prevent heart failure by increasing the production of ATP from the oxidation of ketone bodiesSGLT2i in heart failure patients can reduce blood pressure, maintain a decrease in eGFR and reduce the risk of acute kidney injury (AKI) in patients. SGLT2i is very effective in the treatment of HFrEF patients. Because of these advantages, further research is needed in Indonesia in order to determine the effectiveness and impact of giving SGLT2i to HFrEF patients in Indonesia.

Sodium-glucose cotransporter inhibitors and kidney fibrosis: review of the current evidence and related mechanisms.

Sodium-glucose cotransporter inhibitors (SGLT2i) are a new class of anti-diabetic drugs that have beneficial cardiovascular and renal effects. These drugs decrease proximal tubular glucose reabsorption and decrease blood glucose levels as a main anti-diabetic action. Furthermore, SGLT2i decreases glomerular hyperfiltration by a tubuloglomerular feedback mechanism. However, the renal benefits of these agents are independent of glucose-lowering and hemodynamic factors, and SGLT2i also impacts the kidney structure including kidney fibrosis. Renal fibrosis is a common pathway and pathological marker of virtually every type of chronic kidney disease (CKD), and amelioration of renal fibrosis is of utmost importance to reduce the progression of CKD. Recent studies have shown that SGLT2i impact many cellular processes including inflammation, hypoxia, oxidative stress, metabolic functions, and renin-angiotensin system (RAS) which all are related with kidney fibrosis. Indeed, most but not all studies showed that renal fibrosis was ameliorated by SGLT2i through the reduction of inflammation, hypoxia, oxidative stress, and RAS activation. In addition, less known effects on SGLT2i on klotho expression, capillary rarefaction, signal transducer and activator of transcription signaling and peptidylprolyl cis/trans isomerase (Pin1) levels may partly explain the anti-fibrotic effects of SGLT2i in kidneys. It is important to remember that some studies have not shown any beneficial effects of SGLT2i on kidney fibrosis. Given this background, in the current review, we have summarized the studies and pathophysiologic aspects of SGL2 inhibition on renal fibrosis in various CKD models and tried to explain the potential reasons for contrasting findings.

Considerations for Choosing First-Line Urate-Lowering Treatment in Older Patients with Comorbid Conditions.

Gout is the most common inflammatory arthritis in adults. The prevalence of gout increases with age. Urate-lowering treatment (ULT) among older patients is often challenging in that patients frequently suffer insufficient effectiveness or adverse events due to comorbidities, concurrentmedications, and altered pharmacokinetics. The large-scale randomized controlled trials (RCTs) directly investigating gout patients regarding cardiovascular (CV) safety have only recently been introduced; CARES and FAST compared the CV safety of the two xanthine oxidase inhibitors (XOis), febuxostat versus allopurinol, in patients with gout. Based on the CARES trial that showed CV concerns with febuxostat, the current international guidelines recommend allopurinol as first-line ULT in gout, while preserving other agents as a second-line treatment, despite a higher potency of febuxostat. XOis would be more suitable than uricosurics to treat older patients with gout due to the high prevalence of chronic kidney disease (CKD) in older patients. However, allopurinol alone might not achieve the target serum uric acid levels below 6 mg/dL and CKD might confer an increased risk of allopurinol induced cutaneous adverse reactions in older patients. Furthermore, as well as the later analysis of CARES participants who were lost to follow-up, data from the FAST trial and real-world studies suggest non-inferior CV safety for febuxostat compared to allopurinol even in the presence of CV diseases. Thus, febuxostat use in older patients with renal impairment may be more positively considered. The combination therapy of a novel uricosuric, verinurad, plus febuxostat reduced albuminuria in hyperuricemic patients with type 2 diabetes and CKD in a phase 2a trial, and further RCTs are awaited. Finally, the sodium-glucose cotransporter-2 inhibitor class of oral hypoglycemic agents, known to exert beneficial CV and renal effects independent of glycemic control, have shown a uricosuric effect and could be used as adjunctive therapy in older patients with cardiorenal comorbidities.

Stepwise Strategy to Identify Thrombin as a Hydrolytic Substrate for Nattokinase.

Nattokinase (NK) is a serine protease with a potent thrombolytic activity that possesses multiple cardiovascular disease (CVD) preventative and treatment activities. In light of its advanced beneficial cardiovascular effects and its nature as a serine protease, characterizing its biological substrates is essential for informing and ultimately delineating the molecular mechanism of its thrombolytic and anticoagulant activities that will unlock the powerful strategic design of effective therapies for CVDs. Given the efficacy of NK to break the vicious loop between inflammation, oxidative stress, and thrombosis, and the extensive role of thrombin in the loop, a stepwise computational strategy was developed to investigate the cleavage events of NK, including both a protein-protein complex model for protein substrate recognition and a protease-peptide complex model for the cleavage site identification, whereby their contact region was sited to allow for the prediction of the corresponding cleavage site that was successfully verified by both mass spectrometry (MS)-based N-terminal sequencing and various functional assays. Collectively, thrombin was predicted and identified to be a novel biological substrate of NK, which expanded the comprehensive antithrombus mechanism of NK via breaking the vicious loop between inflammation, oxidative stress, and thrombosis. This study not only provided insight into the interaction characteristics between NK and its hydrolytic substrate for a better understanding toward its catalytic mechanism but also developed a comprehensive computational strategy to elucidate the proteolytic targets of NK for the breakthrough of feature drug development.

Effect of Simvastatin on Hepatic Venous Portal Gradient for Maintenance of Lipid Profile of Liver Cirrhosis patients

Background: Liver cirrhosis has an essential influence on the lipid profile. Statins have well-known beneficial cardiovascular effects reducing cardiovascular events and mortality. Aim: To compare the change in hepatic venous portal gradient with or without simvastatin for maintenance of lipid profile of liver cirrhosis patients Methods: This Randomized controlled trial was done at Department of Medicine, South medical ward, Mayo hospital, Lahore from October 2021 to March 2022.patients were randomly divided in two groups. In group A, patients were given simvastatin with standard treatment. In group B, patients were given standard treatment only. Before and after 1 month of treatment, lipid profile and HVPG level were assessed again. The change in HVPG and lipid profile was calculated. Data was entered and analyzed in SPSS version 25. Results: The mean age of patients received simvastatin was 55.26 ± 13.24 years and mean age of patients in control was 53.74 ± 16.88 years. In simvastatin group, there were 82 (54.7%) males and 68(45.3%) females. In control group, there were 93 (62.0%) males and 57(38%) females. With simvastatin, the mean HVPG level was reduced from 20.11± 6.21mg/dl to 15.42±4.71 mg/dl after a month (mean change = 4.69 ± 1.50 mg/dl). While in control group, mean HVPG level was reduced from 19.28±7.61 mg/dl to 17.82± 6.52 mg/dl after a month (mean change = 1.46±1.09 mg/dl). The effect size was 23.3% with simvastatin while 7.6% without simvastatin for reduction of HVPG in cirrhotic patients. The difference was observed to be significant (p&lt;0.05). Conclusion: Thus, addition of simvastatin is effective in controlling lipid profile of patients with liver cirrhosis. It also has beneficial role in improving condition of liver cirrhosis patients. Keywords: hepatic venous portal gradient, simvastatin, lipid profile, liver cirrhosis, LDL, HDL, total cholesterol, triglycerides

Antioxidant Properties and Beneficial Cardiovascular Effects of a Natural Extract of Pomegranate in Healthy Volunteers: A Randomized Preliminary Single-Blind Controlled Study.

Pomegranates are known to possess anti-hypertensive, anti-atherogenic and cardioprotective effects mainly due to their pleiotropic effects on various cellular pathways, especially those triggered by oxidative stress. The aim of this study was to investigate the effect of natural standardized pomegranate (PE) extract on cardiovascular risk factors in 24 healthy volunteers who participated in a randomized, single-blind placebo-controlled study. There were 12 subjects in the PE group and 12 in the placebo group. Variables were measured at baseline and after 14 and 28 days of supplementation are anthropometry, BP, pulse wave velocity, fat and lean body mass, salivary and urinary cortisol, and cortisone, total phenolics, antioxidant capacity and lipid peroxidation. Urinary total phenolics excretion and antioxidant capacity were significantly increased after 14 and 28 days of PE intake. At day 28, there were also statistically significant decreases in systolic and diastolic blood pressure (BP), pulse wave velocity, body fat and fat mass, as well as an increase in lean body mass. Significant changes in the placebo group were not found. Glucocorticoid levels showed a significant decrease in saliva cortisol at day 28 (morning) in the PE group, and cortisol/cortisone ratio was significantly decreased following 28 days of PE intake at morning, noon, and evening. Urine free cortisol was significantly reduced at day 14. These findings suggest that pomegranate extract intake may improve antioxidant and oxidative stress status and play a beneficial role in the attenuation of some cardiovascular risk factors. Future studies should concentrate on overweight and older people.

GLP-1 receptor expression in human internal thoracic arteries is dependent on low-grade inflammation potentially explaining the cardiovascular protective effect of GLP1-R agonists

Abstract Background Clinical research showed that glucagon-like peptide-1 receptor (GLP1R) agonists (GLP1RA) reduced the incidence of major cardiovascular events in diabetic patients. The protective effect was more noteworthy in patients with established cardiovascular disease, associated with chronic low-grade inflammation resulting in endothelial dysfunction and arterial remodeling and fibrosis. Despite the clinically proven cardiovascular benefit of GLP1RA, the expression pattern of GLP1R and its function in the human vasculature remain poorly studied. Aim This study investigated the expression of GLP1R and its role in internal thoracic arteries (ITA) from patients with coronary artery disease. Methods Human ITA were collected from patients (N=30) undergoing coronary artery bypass surgery at the University Hospital of Strasbourg. Gene expression levels were assessed using RT-qPCR, protein levels by Western blot analysis, in situ tissue localization of proteins by immunofluorescence (IF) staining, and the level of oxidative stress using dihydroethidium. Results ITA showed a substantial difference in GLP1R mRNA levels reaching up to 7-fold among patients. GLP1R mRNA levels were positively correlated with those of SLC5A1, SLC5A2, F3, AT1R, IL1B, IL6, TNFα, VCAM1 and NCF1, and negatively with NOS3 mRNA levels. High GLP1R expressing ITA showed high levels of phosphorylated p65 NF-κB, cell adhesion molecule VCAM1, members of the angiotensin system (ACE1 and AT1R), remodeling and fibrotic markers (MMP9, MMP2, TGFβ) and a low level of eNOS, whereas the contrary was observed for low GLP1R expressing ITA. IF staining showed that GLP1R signals were predominantly observed in the endothelium of low GLP1R expressing ITA and in the vascular smooth muscle of high GLP1R expressing ITA. Prominent CD68 staining associated with the endothelium and the perivascular adipose tissue were observed in high but not low GLP1R expressing ITA. High GLP1R expressing ITA showed increased levels of oxidative stress, which were inhibited by the antioxidant N-acetylcysteine (NAC), NADPH oxidase inhibitor (VAS-2870), ACE inhibitor (perindoprilat), AT1R antagonist (losartan), TNF-α receptor neutralizing antibody (infliximab), selective SGLT2 inhibitor (empagliflozin), dual SGLT1/2 inhibitor (sotagliflozin) and by GLP1R agonists (liraglutide and semaglutide) with inhibitory effects amounting to about 50 to 75%. Conclusion These findings indicate that GLP1R expression levels in human ITA are dependent on low-grade inflammation and linked to endothelial dysfunction, pro-thrombotic, pro-remodeling and pro-fibrotic responses. Furthermore, they were associated with increased level of oxidative stress sensitive to inhibitors of either the local angiotensin system, SGLT2, TNFα and agonists of GLP1R. Thus, the cardiovascular beneficial effects of GLP1R agonists might be attributable to their ability to reduce the pro-oxidant stimulatory signal related to low-grade inflammation. Funding Acknowledgement Type of funding sources: Other. Main funding source(s): Frency Society of Vascular Medicine (SFMV)

Mechanism of canagliflozin-induced vasodilation in resistance mesenteric arteries and the regulation of systemic blood pressure

Canagliflozin, a sodium glucose co-transporter 2 (SGLT2) inhibitor, is reported to produce beneficial cardiovascular effects including a reduction in arterial contractility, and blood pressure. However, whether canagliflozin could directly relax resistance mesenteric arteries, underlying molecular mechanism and its role in regulating systemic blood pressure remain unclear. Here, we investigated the mechanism of regulation of small mesenteric artery contractility and its relevance for blood pressure regulation. Our pressure myography data showed that canagliflozin application rapidly produces a concentration-dependent vasodilation in mesenteric arteries. Such vasodilation was inhibited by concurrent inhibition of smooth muscle cell voltage-gated K+ channels KV1.5 (by 1 μM DPO-1), KV2.1 (by 100 nM guangxitoxin), and KV7 (by 10 μM linopirdine) but not by the inhibition of small-, intermediate-, and large-conductance Ca2+-activated K+ channels (SKCa by 1 μM apamin, IKCa 10 μM TRAM-34, and BKCa by 10 μM paxilline, respectively), ATP-sensitive (KATP) channels (by 10 μM glibenclamide), or SERCA pump (by 0.1 μM thapsigargin). Inhibition of SGLTs (by 1 μM phlorizin or the inhibition of endothelial signaling did not alter canagliflozin-evoked vasodilation. Consistently, acute canagliflozin treatment (4 mg/kg body weight) lowered systemic blood pressure in vivo. Overall, our data suggests that canagliflozin stimulates KV1.5, KV2.1, and KV7 channels, leading to vasodilation and a reduction of systemic blood pressure.

Genetic- and diet-induced ω-3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice.

SUMMARYTRPV4 channel activation in endothelial cells leads to vasodilation, while impairment of TRPV4 activity is implicated in vascular dysfunction. Strategies that increase TRPV4 activity could enhance vasodilation and ameliorate vascular disorders. Here, we show that supplementation with eicosapentaenoic acid (EPA), an ω-3 polyunsaturated fatty acid known to have beneficial cardiovascular effects, increases TRPV4 activity in human endothelial cells of various vascular beds. Mice carrying the C. elegans FAT-1 enzyme, which converts ω-6 to ω-3 polyunsaturated fatty acids, display higher EPA content and increased TRPV4-mediated vasodilation in mesenteric arteries. Likewise, mice fed an EPA-enriched diet exhibit enhanced and prolonged TRPV4-dependent vasodilation in an endothelial cell-specific manner. We also show that EPA supplementation reduces TRPV4 desensitization, which contributes to the prolonged vasodilation. Neutralization of positive charges in the TRPV4 N terminus impairs the effect of EPA on channel desensitization. These findings highlight the beneficial effects of manipulating fatty acid content to enhance TRPV4-mediated vasodilation.

Relationship between echocardiographic characteristics and cardiac biomarkers during long-distance trail running.

BackgroundEven if the beneficial cardiovascular effects of moderate exercise are recognized, effects of prolonged and intense exercise are still debated. This study aims to detect cardiovascular changes associated with long endurance running by assessing the relationship between echocardiographic parameters and cardiac biomarkers during long-distance trail running.MethodsWe performed a prospective observational study that included 20 participants who were all amateur runners (median age of 41 years old, still alive after a 7-year clinical follow-up) from 80-km trail running. All the participants underwent an echocardiographic examination and venous blood sampling before the race, at the intermediate refreshment checkpoints of the race (21st and 53rd km), and within 10 min after arrival.ResultsMitral E/A velocity ratio and mitral TDI e’ wave were significantly decreased at the 21st km to arrival (p < 0.05). Mitral S wave and global longitudinal strain (GLS) were significantly decreased from the 53rd km to arrival (p < 0.05 for 53rd and 80th km). As compared to baseline, T-troponin and NT-proBNP were significantly increased at the 21st km in all the participants, but T-troponin values were systematically increased above the significative threshold. Diastolic echocardiographic abnormalities were mainly observed among participants with highest NT-proBNP (> 77 ng.l–1) values at the 21st km. As compared to baseline, mitral e’ wave was significantly decreased (–35%) in participants with highest values of NT-proBNP. Similarly, GLS was also depressed among participants with highest troponin values at the 53rd km (p = 0.01 for 53rd km and p = 0.04 for arrival).ConclusionDuring the long-distance trail running, the early LV decrease in diastolic echocardiographic parameters is associated with increase in NT pro-BNP blood levels, and the decrease in LV systolic echocardiographic parameters later is associated with increase in T-troponin blood levels.