Aortic Endothelial Dysfunction Research Articles

The role of low-carbohydrate, high-fat diet in modulating autophagy and endoplasmic reticulum stress in aortic endothelial dysfunction of metabolic syndrome animal model.

Endothelial dysfunction (ED) is induced by insulin resistance, mediated by endoplasmic reticulum (ER) stress and disturbed autophagy. This study investigates the protective role of a low-carbohydrate, high-fat (LCHF) diet on ED, ER stress, and autophagy dysregulation in an experimental animal model of metabolic syndrome. Forty male Sprague-Dawley rats were divided into four groups: a Control group (standard diet) and three Dexamethasone (DEX) treated groups. Group II continued the standard diet, Group III received an LCHF diet, and Group IV received a high-carbohydrate, low-fat (HCLF) diet. At the end of the experiment, aortic tissue samples were obtained and used for histological, immunohistochemical (Endothelin and PCNA, biochemical MDA, TCA, NO, 8-OH-dG, and Nrf2/ARE protein) and molecular (Endothelin, eNOS, Nrf-2 α, p62, LC3, BECN-1, PINK1, CHOP, BNIP3, PCNA) analysis. Oxidative stress, autophagy markers, and ED markers are increased in the metabolic syndrome group. LCHF diet mitigates the adverse effects of DEX on endothelial dysfunction and oxidative stress, as evidenced by reduced BMI, HOMA-IR, and improved histological and molecular parameters. Oxidative stress, autophagy dysregulation, and ER stress play crucial roles in the pathogenesis of insulin resistance-induced endothelial dysfunction. An LCHF diet offers protective benefits against insulin resistance and related comorbidities, including endothelial dysfunction.

Effect of esaxerenone on the onset of aortic endothelial dysfunction and circulating microparticles in type 1 diabetic male mice

Endothelial dysfunction exacerbates hypertension and other vascular complications in diabetes mellitus (DM). Circulating microparticles (MPs) and extracellular vesicles released in patients with DM have emerged as novel regulators of endothelial dysfunction. The obstruction of mineralocorticoid receptors (MRs) is a potential therapeutic approach to reduce cardiovascular complications. Their impact on the obstruction of MRs on circulating MPs and endothelial dysfunction in DM remains unclear. DM was induced in mice through a single intravenous dose of streptozotocin (STZ; 200 mg/kg). Esaxerenone (ESAX; 3 mg/kg/day), a MR blocker was administered via diet for 8 weeks. In this study, the aortas of the DM group showed the endothelial dysfunction and the administration of ESAX ameliorated the endothelial-dependent responses. Moreover, ESAX influences the impaired endothelial-dependent responses of DM-derived MPs. Interestingly, MP levels increased in DM whereas decreased after ESAX administration. In the aorta, the DM-derived MPs increased the expression of intercellular adhesion molecule-1 (ICAM-1). ESAX inhibited the adhesion of DM-derived MPs. Moreover, the ICAM-1 inhibitor A205804 shows similar effects as ESAX. These results indicate that the release and adhesion properties of MPs can be partially obstructed by ESAX via the ICAM-1 signaling pathway, which clarifies the other functions beyond the anti-hypertensive effects of ESAX.

Exposure to aircraft noise exacerbates cardiovascular and oxidative damage in three mouse models of diabetes.

Epidemiology links noise to increased risk of metabolic diseases like diabetes and obesity. Translational studies in humans and experimental animals showed that noise causes reactive oxygen species (ROS)-mediated cardiovascular damage. The interaction between noise and diabetes, specifically potential additive adverse effects, remains to be determined. C57BL/6 mice were treated with streptozotocin (i.p. injections, 50 mg/kg/d for 5d) to induce type-1 diabetes, with S961 (subcutaneous osmotic minipumps, 0.57 mg/kg/d for 7d) or fed a high-fat diet (HFD, 20 weeks) to induce type-2 diabetes. Control and diabetic mice were exposed to aircraft noise to an average sound pressure level of 72 dB(A) for 4d. While body weight was unaffected, noise reduced insulin production in all diabetes models. The oral glucose tolerance test showed only an additive aggravation by noise in the HFD model. Noise increased blood pressure and aggravated diabetes-induced aortic, mesenteric, and cerebral arterioles endothelial dysfunction. ROS formation in cerebral arterioles, the aorta, the heart, and isolated mitochondria was consistently increased by noise in all models of diabetes. Mitochondrial respiration was impaired by diabetes and noise, however without additive effects. Noise increased ROS and caused inflammation in adipose tissue in the HFD model. RNA sequencing data and alteration of gene pathway clusters also supported additive damage by noise in the setting of diabetes. In all three models of diabetes, aircraft noise exacerbates oxidative stress, inflammation, and endothelial dysfunction in mice with pre-existing diabetes. Thus, noise may potentiate the already increased cardiovascular risk in diabetic patients.

The Role of Amino Acid Glycine on Cardiovascular Health and Its Beneficial Effects: A Narrative Review

Glycine, a simple amino acid, is not only essential due to its potential insufficiency in vivo, but also has significant metabolic functions. It serves as a crucial building block for proteins. At the same time, as a bioactive molecule, it regulates gene expression for cytoprotection, protein configuration and activity, and other critical biological processes, including glutathione synthesis. The intriguing, beneficial effects of glycine in medical applications have piqued the research community’s interest in recent decades. This work delves into the compelling discoveries about the pivotal role of glycine in cardiovascular health and its intricate mechanisms of action for alleviating several medical conditions. Glycine’s broad spectrum of impact spans numerous diseases, encompassing not only acute myocardial infarction, aortic dissection, and cardiac hypertrophy, but also transplant rejections of aortic allografts, insulin resistance, and endothelial dysfunction, thereby providing a comprehensive understanding of its medical applications.

The Circulating Milieu Mediates Arterial Dysfunction with Aging: Protection by Habitual Aerobic Exercise

Age-related arterial dysfunction (aortic stiffening and endothelial dysfunction) is the key antecedent to cardiovascular diseases. Habitual aerobic exercise largely mitigates arterial dysfunction in mid-life/older (ML/O) adults; however, the mechanisms by which this occurs remain to be fully elucidated. The circulating milieu (bioactive factors in the bloodstream) shifts towards a pro-inflammatory/oxidative state with advancing age but is preserved with habitual aerobic exercise. However, it is unclear if the circulating milieu directly contributes to age-related arterial dysfunction and if habitual aerobic exercise preserves arterial function with aging, in part, by favorably modulating the circulating milieu. PURPOSE: To determine the influence of aging and habitual aerobic exercise on circulating milieu-mediated aortic stiffening and endothelial function. METHODS: Human serum was collected from healthy young (18-29 yr) and ML/O (50+ yr) men and women. Aerobic exercise status was determined by leisure-time physical activity (>60 h/wk). Aortic rings (~1 mm) and common carotid arteries were obtained from young adult (3-4 mo), intervention naïve male and female C57BL/6N mice to assess aortic stiffness and endothelial function, respectively. Arteries were exposed ex vivofor 24-48 hours to 5% sex-matched human serum from one of the following groups: young sedentary (n=9; 4F), ML/O sedentary (n=14; 7F) or ML/O habitually aerobic exercise trained (n=8; 4F). Aortic stiffness was measured as elastic modulus (a measure of intrinsic mechanical wall stiffness). Endothelial function was assessed as endothelium-dependent dilation (EDD) and endothelium-independent dilation (EID) as peak response to acetylcholine and the nitric oxide (NO) donor sodium nitroprusside, respectively. Differences were determined via one-way ANOVA. RESULTS: Aortic Stiffness. Aortic elastic modulus was higher in aorta rings incubated with ML/O sedentary serum compared to young sedentary serum (ML/O sedentary, 4912 ± 588 v. Young sedentary, 3019 ± 522 kPa; p=0.020). Compared with ML/O sedentary serum, aorta rings incubated with ML/O trained serum had lower elastic modulus (2237 ± 426 kPa, p=0.002) which was not different to aorta rings incubated with young sedentary serum (p=0.369). Endothelial Function. Peak EDD was lower in carotid arteries incubated with ML/O sedentary v. young sedentary serum (79 ± 3% v. 92 ± 2%, p=0.001). Compared with ML/O sedentary serum, incubation with ML/O trained serum evoked higher peak EDD (93 ± 1%; p=0.001) that was similar to that produced by Young sedentary serum (p=0.866). There were no group differences in peak EID (p=0.228) suggesting that circulating milieu-induced differences were endothelium-specific and were not attributable to changes in smooth muscle sensitivity to NO. CONCLUSION: These findings suggest that the circulating milieu is a key mechanism by which aging induces arterial dysfunction and by which habitual aerobic exercise training preserves arterial function with aging. F31 HL165885, R21 AG078408, K99 HL159241, K99/R00 HL151818. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Early life stress-induced aortic endothelial dysfunction is mediated by endothelium-specific HDAC9

Early life stress (ELS) is highly prevalent worldwide and has been associated with an elevated risk of developing cardiovascular disease (CVD) in adulthood. We previously showed that maternal separation with early weaning (MSEW), an established ELS model of neglect in mice, leads to blunted endothelium-dependent relaxation as well as increased histone deacetylase (HDAC) activity. We hypothesized that specific HDAC isoform(s) mediate the ELS-induced endothelial dysfunction. In a screen of all HDAC isoforms, HDAC1, 6, and 9 gene expression were significantly elevated in aortas from male adult MSEW and normally reared (NR) C57BL6/J mice (N=3/group, p<0.05). Western blot analysis found that only HDAC9 protein abundance was significantly elevated in aortas from male MSEW as compared to NR mice (N=6/group, p=0.01). Detection of HDAC9 via immunohistochemistry was greater in the aortic endothelium of MSEW mice compared to NR (N=5/group). Subsequently, vascular reactivity was measured with wire myography on tamoxifen inducible endothelium-specific HDAC9KO (Cdh5CreERT2-HDAC9) MSEW and NR adult mice to determine the role of HDAC9 in MSEW-induced endothelial dysfunction. Male mice exposed to MSEW displayed blunted aortic endothelial-dependent relaxation to acetylcholine compared to NR, whereas endothelial-specific knockout of HDAC9 (HDAC9KO) restored endothelial-dependent relaxation (NR: Emax=89.27%±4.15, N=5; MSEW: Emax=48.12%±22.12, N=2; NR HDAC9 KO: Emax=68.98%±4.73, N=5, MSEW HDAC9KO: Emax=70.50%±1.85, N=3). Females demonstrated no difference between MSEW and NR endothelial dependent relaxation, however, HDAC9KO MSEW displayed blunted relaxation compared to all other groups (NR: Emax= 77.14%±4.86, N=5; MSEW: Emax=84.30%±8.71, N=2; NR HDAC9 KO: Emax=70.80%±5.87, N=5, MSEW HDAC9KO: Emax=69.76%±9.10, N=4). Endothelial-independent relaxation to sodium nitroprusside (SNP) was similar in all groups in both sexes (Male: NR: Emax=96.92%±1.26, N=5; MSEW: Emax=100%±0, N=2; NR HDAC9 KO: Emax=96.52%±3.484, N=5; MSEW HDAC9KO: Emax=100%±0, N=3; Female: NR: Emax=98.58%±0.42, N=5; MSEW: Emax=100%±0, N=2; NR HDAC9 KO: Emax=98.48%±1.08, N=5; MSEW HDAC9KO: Emax=99.15%±0.85, N=4). These findings suggest that HDAC9 mediates ELS-induced endothelial dysfunction in males, however, HDAC9 may play a protective role in females. Future studies aim to further investigate sex-dependent differences in the specific role of HDAC9 in ELS related CVD. NIH P01 HL158500, F31HL165863-01, T32GM135028. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Proximal convoluted tubule cells-derived exosomes attenuate aging-related vascular dysfunction

Aging is an identified risk factor for cardiovascular disease. Aging is associated with increased prevalence of endothelial dysfunction and arterial stiffening. Exosomes are bi-lipid nanoscale vesicles that play important roles in intercellular communications. The purpose of this study was to determine whether administration of renal proximal convoluted tubule cells (PCT)-derived exosomes affects vascular and renal function in aging mice. Aging (24 months old) and young mice (3 months old) were treated with PCT-derived exosomes via non-invasive injection into the kidney 3 times a week for 2 weeks. Administration of PCT-derived exosomes did not affect blood pressure or urinary Na+, K+, and Ca2+ excretions, indicating that PCT-derived exosomes did not affect kidney function. Pulse wave velocity (PWV) was increased in aging mice, indicating arterial stiffening. Interestingly, treatment with PCT-exosomes effectively reduced aging-associated arterial stiffening. In addition, aortic endothelium-dependent relaxing (EDR) responses to acetylcholine (Ach) were reduced in aging mice, indicating endothelial dysfunction. Treatment with PCT-derived exosomes rescued aging-associated aortic endothelial dysfunction. Mechanistically, aortic expression of VCAM1, ICAM1, TNFα, and MCP-1 was upregulated in aging mice, indicating vascular inflammation. Notably, aging-associated vascular inflammation was largely attenuated by delivery of PCT-derived exosomes. Our results suggest that PCT-derived exosomes improves aging-associated arterial stiffening and vascular dysfunction partly via inhibiting vascular inflammation. The findings may help develop novel therapeutic strategies for aging-associated vascular dysfunction using PCT-derived exosomes. This work is supported by NIH research grants R01 HL154147 and AG062375. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Fish oil alleviates diabetes-induced aortic endothelial dysfunction and injuries in mice

Diabetes mellitus (DM)-induced endothelial dysfunction (DED) contributes to macrovascular complications of DM, resulting in high mortality. Fish oil (FO) is a dietary source of omega-3 polyunsaturated fatty acids, with little known for its action on DED. Thus, the objective of the current study was to investigate the effect of FO on aortic DED using a mouse model of DM. To this end, C57BL/6 J male mice (n = 8/each group) were intraperitoneally injected with streptozotocin (50 mg/kg/day, 5 days), following feeding with a high-fat diet containing FO for 24 weeks, with fasting blood glucose levels and glucose tolerance monitored. At the end of the procedure, aortic rings were harvested and evaluated for the contraction and relaxation in response to phenylephrine and acetylcholine respectively. Aortic pathological injuries were assessed by hematoxylin and eosin, and Masson's trichrome staining. Aortic expression of proinflammatory, oxidative stress and profibrotic markers were determined by qRT-PCR and immunohistochemical staining. The diabetic mice developed impaired glucose tolerance, abnormal endothelial dependent aortic contraction and relaxation, thickened tunica media, and increased accumulation of fibrosis in the aortas. These effects were remarkably reversed by FO supplementation. Further investigation revealed that FO inhibited aortic mRNA expression of Tnf-α, Il-6, Il-1β and Vcam-1 as markers for inflammation, Nox1, Nox2, Nox4 and Nos2 as markers for oxidative stress, as well as Acta 2, Col1a1, Col3 and Ccn2 as markers for fibrosis. Moreover, FO decreased the aortic protein expression of TNF-α, 4-HNE and COL1, all of which were increased under the diabetic condition. The present study may provide a molecular basis for the application of FO in prevention of macrovascular complications of DM.

Sodium valproate treatment reverses endothelial dysfunction in aorta from rabbits with acute myocardial infarction

Sodium valproate (VPA), a histone deacetylase (HDAC) inhibitor, could be a promising candidate to treat acute myocardial infarction (AMI). In this study, AMI was induced in New Zealand White rabbits by occluding the left circumflex coronary artery for 1 h, followed by reperfusion. The animals were distributed into three experimental groups: the sham-operated group (SHAM), the AMI group and the AMI + VPA group (AMI treated with VPA 500 mg/kg/day). After 5 weeks, abdominal aorta was removed and used for isometric recording of tension in organ baths or protein expression by Western blot, and plasma for the determination of nitrate/nitrite (NOx) levels by colorimetric assay. Our results indicated that AMI induced a reduction of the endothelium-dependent response to acetylcholine without modifying the endothelium-independent response to sodium nitroprusside, leading to endothelial dysfunction. VPA treatment reversed AMI-induced endothelial dysfunction and even increased NO sensitivity in vascular smooth muscle. This response was consistent with an antioxidant effect of VPA, as it was able to reverse the superoxide dismutase 1 (SOD 1) down-regulation induced by AMI. Our experiments also ruled out that the VPA mechanism was related to eNOS, iNOS, sGC and arginase expression or changes in NOx plasma levels. Therefore, we conclude that VPA improves vasodilation by increasing NO bioavailability, likely due to its antioxidant effect. Since endothelial dysfunction was closely related to AMI, VPA treatment could increase aortic blood flow, making it a potential agent in reperfusion therapy that can prevent the vascular damage.

(065) Characterization of Cardiovascular Function, Metabolic Status and Erectile Function in Preclinical Model of Cardiometabolic Syndrome

Abstract Introduction Cardiometabolic syndrome is a widespread health issue and common cause of erectile dysfunction (ED). Cardiometabolic syndrome encompasses a group of indications including abdominal obesity, insulin-resistant glucose metabolism, dyslipidemia, and hypertension. To date, studies have looked at high fructose or high fat diets to induce cardiometabolic syndrome and ED in rodents, but these models do not closely model the clinical phenotype. Furthermore, the molecular basis of ED in cardiometabolic syndrome needs to be elucidated. Objective Our objective is to characterize the cardiovascular and erectile function, and metabolic status in the ZSF-1 obese rat. This hybrid rat is a cross between a Zucker diabetic fatty female and a spontaneously hypertensive heart failure male rat. We hypothesize that with age the ZSF-1 obese rats will demonstrate similar disease phenotype as seen clinically and develop ED. Methods We used male lean and obese ZSF-1 rats (n=8/group). We measured body weight weekly, and performed apomorphine erectile testing, MRI to assess body composition and glucose tolerance tests at 10, 20, 30 weeks. At 30 weeks, we assessed erectile function using ICP/MAP and vascular stiffness was measured via laser doppler pulse wave velocity (PWV). Additionally, acetylcholine (Ach)-mediated vasodilation, vasodilation to sodium nitroprusside (SNP), and contraction to phenylephrine (PE) was measured in thoracic and abdominal aortas. Aorta and penile segments were collected for histological analyses. Results On average, obese rats were 60% larger (230g) than controls, had greater percent body fat and had significantly impaired glucose tolerance at all time points (p<0.05). Over time, ZSF-1 obese rats developed ED as there was a significant decrease in apomorphine-induced erections by 30 weeks (p<0.05). ED was confirmed by significantly reduced ICP/MAP in ZSF-1 obese compared to lean rats (p<0.05). Obese rats were significantly more hypertensive compared to lean rats (MAP: 140.2 mmHg vs 99.6mmHg, p<0.05). Lean and obese rats had no difference in vascular stiffness indicated by PWV. In our tissue bath experiments, ACh-mediated vasodilation was significantly decreased in the thoracic aorta of obese rats (p<0.05) while the abdominal aorta was unchanged. SNP-mediated vasodilation was similar between groups in both thoracic and abdominal aortas. At high concentrations, PE-mediated vasoconstriction was significantly increased in the thoracic and decreased in the abdominal aortas of obese rats (p<0.05). Conclusions To our knowledge, this is the first report on the erectile function in the ZSF-1 rat model of cardiometabolic syndrome. The ZSF-1 obese rats had marked adiposity, hypertension, hyperglycemia and presented with aortic endothelial dysfunction and ED. We are currently assessing testosterone levels and penile fibrosis. We believe that the ZSF-1 rat is useful for studying ED as they gradually develop metabolic disease and ED over time like the human disease. Disclosure No.

The role of miR-134-5p in 7-ketocholesterol-induced human aortic endothelial dysfunction.

Atherosclerotic cardiovascular diseases are the leading causes of morbidity and mortality worldwide. In our previous study, a panel of miRNA including miR-134-5p was deregulated in young acute coronary syndrome (ACS) patients. However, the roles of these ACS-associated miRNAs in endothelial dysfunction, an early event preceding atherosclerosis, remain to be investigated. In the present study, human aortic endothelial cells (HAECs) were treated with 7-ketocholesterol (7-KC) to induce endothelial dysfunction. Following treatment with 20 μg/ml 7-KC, miR-134-5p was significantly up-regulated and endothelial nitric oxide synthase (eNOS) expression was suppressed. Endothelial barrier disruption was evidenced by the deregulation of adhesion molecules including the activation of focal adhesion kinase (FAK), down-regulation of VE-cadherin, up-regulation of adhesion molecules (E-selectin and ICAM-1), increased expression of inflammatory genes (IL1B, IL6 and COX2) and AKT activation. Knockdown of miR-134-5p in 7-KC-treated HAECs attenuated the suppression of eNOS, the activation of AKT, the down-regulation of VE-cadherin and the up-regulation of E-selectin. In addition, the interaction between miR-134-5p and FOXM1 mRNA was confirmed by the enrichment of FOXM1 transcripts in the pull-down miRNA-mRNA complex. Knockdown of miR-134-5p increased FOXM1 expression whereas transfection with mimic miR-134-5p decreased FOXM1 protein expression. In summary, the involvement of an ACS-associated miRNA, miR-134-5p in endothelial dysfunction was demonstrated. Findings from this study could pave future investigations into utilizing miRNAs as a supplementary tool in ACS diagnosis or as targets for the development of therapeutics.

Protective effect of microbiota-derived short chain fatty acids on vascular dysfunction in mice with systemic lupus erythematosus induced by toll like receptor 7 activation

Our objective was to investigate whether short-chain fatty acids (SCFAs), specifically acetate and butyrate, could prevent vascular dysfunction and elevated blood pressure (BP) in mice with systemic lupus erythematosus (SLE) induced by TLR7 activation using imiquimod (IMQ). Treatment with both SCFAs and dietary fibers rich in resistant starch (RS) or inulin-type fructans (ITF) effectively prevented the development of hypertension and cardiac hypertrophy. Additionally, these treatments improved aortic relaxation induced by acetylcholine and mitigated vascular oxidative stress. Acetate and butyrate treatments also contributed to the maintenance of colonic integrity, reduced endotoxemia, and decreased the proportion of helper T (Th)17 cells in mesenteric lymph nodes (MLNs), blood, and aorta in TLR7-induced SLE mice. The observed changes in MLNs were correlated with increased levels of GPR43 mRNA in mice treated with acetate and increased GPR41 levels along with decreased histone deacetylase (HDAC)− 3 levels in mice treated with butyrate. Notably, the effects attributed to acetate, but not butyrate, were nullified when co-administered with the GPR43 antagonist GLPG-0974. T cell priming and differentiation into Th17 cells in MLNs, as well as increased Th17 cell infiltration, were linked to aortic endothelial dysfunction and hypertension subsequent to the transfer of faecal microbiota from IMQ-treated mice to germ-free (GF) mice. These effects were counteracted in GF mice through treatment with either acetate or butyrate. To conclude, these findings underscore the potential of SCFA consumption in averting hypertension by restoring balance to the interplay between the gut, immune system, and vascular wall in SLE induced by TLR7 activation.

Circ_0003575 knockdown alleviates ox-LDL-induced human aortic endothelial cell dysfunction in atherosclerosis by miR-637/TRAF6 axis.

Circular RNAs (circRNAs) are involved in the progression of atherosclerosis (AS). The present study aimed to determine the functions and mechanism of circ_0003575 in AS. Oxidized low-density lipoprotein (ox-LDL) was used to induce human aortic endothelial cells (HAECs) to establish an AS cell model. Cell Counting Kit-8 (CCK-8) assay and 5'-ethynyl-2'-deoxyuridine (EdU) assay were conducted to assess cell proliferation. Flow cytometry analysis was utilized to quantify cell apoptosis. Tube formation assay was performed to analyze angiogenesis ability. Enzyme linked immunosorbent assay (ELISA) was used to examine the concentrations of inflammatory factors. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were manipulated for the expression of circ_0003575, microRNA-637 (miR-637) and TNF receptor associated factor 6 (TRAF6). Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were adopted to estimate the downstream targets of circ_0003575. Ox-LDL treatment repressed the proliferation and angiogenesis and promoted the apoptosis and inflammation in HAECs. Circ_0003575 knockdown ameliorated ox-LDL-induced injury of HAECs. Circ_0003575 interacted with mi-R-637, which directly targeted TRAF6. Inhibition of miR-637 reversed the impacts of circ_0003575 knockdown on HAEC injury. Moreover, miR-637 overexpression promoted cell proliferation and angiogenesis and inhibited cell apoptosis and inflammation by targeting TRAF6 in ox-LDL-treated HAECs. Further, circ_0003575 silencing inhibited the activation of NF-κB pathway. Circ_0003575 knockdown alleviated ox-LDL-induced HAEC damage by regulating miR-637/TRAF6 and NF-κB pathways.

Targeting the gut microbiota with dietary fibers: a novel approach to prevent the development cardiovascular complications linked to systemic lupus erythematosus in a preclinical study

ABSTRACT This study is to investigate whether dietary fiber intake prevents vascular and renal damage in a genetic mouse model of systemic lupus erythematosus (SLE), and the contribution of gut microbiota in the protective effects. Female NZBWF1 (SLE) mice were treated with resistant-starch (RS) or inulin-type fructans (ITF). In addition, inoculation of fecal microbiota from these experimental groups to recipient normotensive female C57Bl/6J germ-free (GF) mice was performed. Both fiber treatments, especially RS, prevented the development of hypertension, renal injury, improved the aortic relaxation induced by acetylcholine, and the vascular oxidative stress. RS and ITF treatments increased the proportion of acetate- and butyrate-producing bacteria, respectively, improved colonic inflammation and integrity, endotoxemia, and decreased helper T (Th)17 proportion in mesenteric lymph nodes (MLNs), blood, and aorta in SLE mice. However, disease activity (splenomegaly and anti-ds-DNA) was unaffected by both fibers. T cell priming and Th17 differentiation in MLNs and increased Th17 infiltration was linked to aortic endothelial dysfunction and hypertension after inoculation of fecal microbiota from SLE mice to GF mice, without changes in proteinuria and autoimmunity. All these effects were lower in GF mice after fecal inoculation from fiber-treated SLE mice. In conclusion, these findings support that fiber consumption prevented the development of hypertension by rebalancing of dysfunctional gut-immune system-vascular wall axis in SLE.

Cellular Senescence Contributes to Large Elastic Artery Stiffening and Endothelial Dysfunction With Aging: Amelioration With Senolytic Treatment.

Here, we assessed the role of cellular senescence and the senescence associated secretory phenotype (SASP) in age-related aortic stiffening and endothelial dysfunction. We studied young (6-8 mo) and old (27-29 mo) p16-3MR mice, which allows for genetic-based clearance of senescent cells with ganciclovir (GCV). We also treated old C57BL/6N mice with the senolytic ABT-263. In old mice, GCV reduced aortic stiffness assessed by aortic pulse wave velocity (PWV; 477±10 vs. 382±7 cm/s, P<0.05) to young levels (old-GCV vs. young-vehicle, P=0.35); ABT-263 also reduced aortic PWV in old mice (446±9 to 356±11 cm/s, P<0.05). Aortic adventitial collagen was reduced by GCV (P<0.05) and ABT-263 (P=0.12) in old mice. To show an effect of the circulating SASP, we demonstrated that plasma exposure from Old-vehicle p16-3MR mice, but not from Old-GCV mice, induced aortic stiffening assessed ex vivo (elastic modulus; P<0.05). Plasma proteomics implicated glycolysis in circulating SASP-mediated aortic stiffening. In old p16-3MR mice, GCV increased endothelial function assessed via peak carotid artery endothelium-dependent dilation (EDD; Old-GCV, 94±1% vs. Old-vehicle, 84±2%, P<0.05) to young levels (Old-GCV vs. young-vehicle, P=0.98), and EDD was higher in old C57BL/6N mice treated with ABT-263 vs. vehicle (96±1% vs. 82±3%, P<0.05). Improvements in endothelial function were mediated by increased nitric oxide (NO) bioavailability (P<0.05) and reduced oxidative stress (P<0.05). Circulating SASP factors related to NO signaling were associated with greater NO-mediated EDD following senescent cell clearance. Cellular senescence and the SASP contribute to vascular aging and senolytics hold promise for improving age-related vascular function.

Abstract P2146: Endothelial Cell Specific Senescent Cell Clearance Alleviated Vascular Stiffness And Metabolic Dysfunction In Obese Mice

Aging is the leading predictor of acquired cardiovascular disease (CVD), and emerging evidence suggests cellular senescence plays a causal role in development of clinically-significant CVD. While we previously reported that senescent cell elimination delays or prevents progression of several aging phenotypes in aging mice, the responsible types of senescent cells contributing to cardiovascular dysfunction in vivo are not yet elucidated. To test the hypothesis that senescent endothelial cells are a major contributor to cardiovascular dysfunction, we developed Tie2-Cre; p16 Ink4a - LOX-ATTAC mice, which allows for specific elimination of p16 Ink4a+ senescent endothelial cells upon administration of AP20187 (which only affects cells carrying the conditionally-expressed ATTAC construct). Using a mouse model of diabetic vascular disease (40% fat diet for 12 weeks , HFD), we found that biweekly selective removal of senescent endothelial cells alleviated HFD-induced endothelial dysfunction in aorta (assessed ex vivo using isolated organ baths). Additionally, ejection fraction (assessed using in vivo echocardiography) was preserved in AP20187-treated mice (driving ATTAC-dependent removal of senescent endothelial cells) compared to their vehicle-treated controls (76.6%±4.2, 63.2%±4.2). In addition to improvement in cardiovascular function, metabolic parameters (HbA1c and fasting glucose levels) were improved by senescent endothelial cell elimination. Collectively, these data suggest that elimination of p16 Ink4a+ senescent endothelial cells may be a viable therapeutic strategy for alleviation of metabolic disease-induced cardiovascular dysfunction.

Waterpipe smoke inhalation induces lung injury and aortic endothelial dysfunction in mice.

Waterpipe tobacco smoking (WPS) inhalation has been shown to trigger endothelial dysfunction and atherosclerosis. However, the mechanisms underlying these effects are still unknown. Here, we assessed the impact and underlying mechanism of WPS exposure for one month on endothelial dysfunction using aortic tissue of mice. The duration of the session was 30 min/day and 5 days/week. Control mice were exposed to air. Inhalation of WPS induced an increase in the number of macrophages and neutrophils and the concentrations of protein, tumor necrosis factor alpha (TNF alpha), interleukin (IL)-1beta, and glutathione in bronchoalveolar lavage fluid. Moreover, the concentrations of proinflammatory cytokines (TNF alpha, IL-6 and IL-1beta), adhesion molecules (intercellular adhesion molecule-1, vascular cell adhesion molecule-1, E-selectin and P-selectin) and markers of oxidative stress (lipid peroxidation, glutathione, superoxide dismutase and nitric oxide) in aortic homogenates of mice exposed to WPS were significantly augmented compared with air exposed mice. Likewise, the concentration of galectin-3 was significantly increased in the aortic homogenates of mice exposed to WPS compared with control group. WPS inhalation induced vascular DNA damage assessed by comet assay and apoptosis characterized by a significant increase in cleaved caspase-3. While the aortic expression of phosphorylated nuclear factor kappaB (NF-kappaB) was significantly increased following WPS inhalation, the concentration of sirtuin 1 (SIRT1) was significantly decreased in WPS group compared with air-exposed group. In conclusion, our study provided evidence that WPS inhalation triggers lung injury and endothelial inflammation, oxidative stress and apoptosis which were associated with nuclear factor-kappaB activation and SIRT1 down-regulation.

Mitigation effect of galangin against aortic dysfunction and hypertrophy in rats with metabolic syndrome

Vascular alterations induced by a high-fat diet (HFD) are involved in the development of hypertension. Galangin, a flavonoid, is the major active compound isolated from galangal and propolis. The objective of this study was to investigate the effect of galangin on aortic endothelial dysfunction and hypertrophy, and the mechanisms involved in HFD-induced metabolic syndrome (MS) in rats. Male Sprague-Dawley rats (220–240 g) were separated into three groups: control + vehicle, MS + vehicle, and MS + galangin (50 mg/kg). Rats with MS received HFD plus 15% fructose solution for 16 weeks. Galangin or vehicle was orally administered daily for the final four weeks. Galangin reduced body weight and mean arterial pressure in HFD rats (p < 0.05). It also reduced circulating fasting blood glucose, insulin, and total cholesterol levels (p < 0.05). Impaired vascular responses to the exogenous acetylcholine observed in the aortic ring of HFD rats were restored by galangin (p < 0.05). However, the response to sodium nitroprusside did not differ between the groups. Galangin enhanced the expression of the aortic endothelial nitric oxide synthase (eNOS) protein and increased circulating nitric oxide (NO) levels in the MS group (p < 0.05). Aortic hypertrophy in HFD rats was alleviated by galangin (p < 0.05). Increases in tumour necrosis factor-alpha (TNF-α), interleukin (IL)-6 levels, angiotensin-converting enzyme activity and angiotensin II (Ang II) concentrations in rats with MS were suppressed in galangin treated group (p < 0.05). Furthermore, galangin reduced the upregulation of angiotensin II type I receptor (AT1R) and transforming growth factor-beta (TGF-β) expression in rats with MS (p < 0.05). In conclusion, galangin alleviates metabolic disorders and improves aortic endothelial dysfunction and hypertrophy in the MS group. These effects were consistent with increased NO availability, reduced inflammation, and suppressing Ang II/AT1R/TGF-β signalling pathway.

TUDCA treatment restores early protein-restriction-induced aortic hipocontractility and endothelial dysfunction

TUDCA treatment restores early protein-restriction-induced aortic hipocontractility and endothelial dysfunction Israelle Netto Freitas1,2; Joel da Silva Junior2; Jamaira Aparecida Victorio1,2; Everardo Magalhães Carneiro1,2; Ana Paula Davel1 1Laboratory of Vascular Biology and 2Obesity and Comorbidities Center-OCRC, Institute of Biology, University of Campinas-UNICAMP, Campinas, SP, Brazil. Malnutrition is a risk factor for the development of vascular dysfunction. Tauroursodeoxycholic acid (TUDCA) inhibits endoplasmic reticulum stress (ERS) and has been demonstrated to be protective against vascular dysfunction in cardiometabolic diseases. Therefore, we hypothesized that TUDCA could be beneficial for endothelial function and vascular contractility during malnutrition. For this, post-weaning male and female mice fed a normoprotein (NP, 14% protein) or low protein (LP, 6% protein, isocaloric) diet for 15 weeks. In the last 2 weeks, some NP and LP animals received TUDCA (300 mg/kg/day, ip) or vehicle. At the end of the treatment, body weight (BW) and systolic blood pressure (SBP) were evaluated and thoracic aortic isolated to assess vascular responses, fluorescence to dihydroethidium (DHE) and gene expression. As results, BW was reduced and SBP was elevated in male and female protein-restricted groups; TUDCA treatment restored SBP but not BW. Only in males, protein restriction impaired endothelium-dependent relaxation to acetylcholine and reduced contractility (phenylephrine, serotonin, and TXA2 analogue U46619) in aorta. TUDCA reversed both endothelial dysfunction and hypocontractility in LP male group. We found reduced expression of smooth muscle 22α (SM22α), α-actin and L-type calcium channel (Cav1.2) in aorta from male LP compared to NP, which was restored by TUDCA treatment. In addition, aorta from male LP group showed increased DHE fluorescence and gene expression of the ERS markers GRP78, ATF6, PERK, IRE1α and CHOP. These alterations were reversed by TUDCA. In conclusion, our data suggest that post-weaning protein restriction results in endothelial dysfunction and aortic hypocontractility in males only. These alterations were associated with oxidative stress and reduced expression of vascular contractile and ERS markers. TUDCA treatment rescued vascular contraction and endothelial function in early protein-restricted mice evidencing the therapeutic potential of this bile acid for the vascular complications associated with undernutrition. The authors have nothing to disclose. Funding source: São Paulo Research Foundation (FAPESP). This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Reactive Oxygen Species in the Aorta and Perivascular Adipose Tissue Precedes Endothelial Dysfunction in the Aorta of Mice with a High-Fat High-Sucrose Diet and Additional Factors.

Metabolic syndrome (Mets) is the major contributor to the onset of metabolic complications, such as hypertension, type 2 diabetes mellitus (DM), dyslipidemia, and non-alcoholic fatty liver disease, resulting in cardiovascular diseases. C57BL/6 mice on a high-fat and high-sucrose diet (HFHSD) are a well-established model of Mets but have minor endothelial dysfunction in isolated aortas without perivascular adipose tissue (PVAT). The purpose of this study was to evaluate the effects of additional factors such as DM, dyslipidemia, and steatohepatitis on endothelial dysfunction in aortas without PVAT. Here, we employed eight-week-old male C57BL/6 mice fed with a normal diet (ND), HFHSD, steatohepatitis choline-deficient HFHSD (HFHSD-SH), and HFHSD containing 1% cholesterol and 0.1% deoxycholic acid (HFHSD-Chol) for 16 weeks. At week 20, some HFHSD-fed mice were treated with streptozocin to develop diabetes (HFHSD-DM). In PVAT-free aortas, the endothelial-dependent relaxation (EDR) did not differ between ND and HFHSD (p = 0.25), but in aortas with PVAT, the EDR of HFHSD-fed mice was impaired compared with ND-fed mice (p = 0.005). HFHSD-DM, HFHSD-SH, and HFHSD-Chol impaired the EDR in aortas without PVAT (p < 0.001, p = 0.019, and p = 0.009 vs. ND, respectively). Furthermore, tempol rescued the EDR in those models. In the Mets model, the EDR is compromised by PVAT, but with the addition of DM, dyslipidemia, and SH, the vessels themselves may result in impaired EDR.