bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mechanisms coordinating vascular smooth muscle adhesion to the extracellular matrix during contraction
Authors: Huang Huang , Sun Zhe , Hill Michael A , Meininger Gerlad A. 
Published: November 2023

Physical coupling of vascular smooth muscle cells (VMSC) to extracellular matrix proteins (ECM) by integrins is required to efficient mechano‐transduction and vessel function. Previous work in our laboratory demonstrated that increased VSMC stiffness, induced by Angiotensin II (AII), is coordinated with enhanced adhesion to fibronectin (FN). However, the mechanism of coordination is not fully understood. In this study, we hypothesized that enhanced adhesion in response to AII was mediated by integrin activation through an inside‐out signaling pathway linked to increases in Ca2+. We first tested that the increase in FN adhesion was due to integrin activation and not increased membrane expression. We found that the adhesion, studied, using atomic force microscopy (AFM) with anti‐alpha 5 integrin‐coated probes, was not increased by AII in contrast to increased adhesion observed with FN coated probes. This suggested that the enhanced adhesion was occurring through an integrin activation process and not an increase in membrane density of the integrin. To examine a role for Ca2+ in this integrin activation, we used a non‐G‐protein coupled vasoconstrictor KCl (60mM), and assessed the coordination between changes in cell elasticity and adhesion to FN. In all VSMC tested, cell stiffness was increased by KCl, and this enhanced stiffness was transient and rapid. Also, cell adhesion probability (number of adhesion events) was transiently increased by KCl but the average adhesion force was unchanged. These data suggest that Ca2+ may play a role in the activation of integrin adhesion during contraction. However, further studies are necessary to confirm a role for Ca2+ and elucidate the signaling pathway linked to integrin activation. d adhesion to FN. In all VSMC tested, cell stiffness was increased by KCl, and this enhanced stiffness was transient and rapid. Also, cell adhesion probability (number of adhesion events) was transiently increased by KCl but the average adhesion force was unchanged. These data suggest that Ca2+ may play a role in the activation of integrin adhesion during contraction. However, further studies are necessary to confirm a role for Ca2+ and elucidate the signaling pathway linked to integrin activation.Support or Funding InformationNIH 1PO1HL095486 to GAM
Human pericardial fluid‐derived endothelin elicits arterial constriction
Authors: Nemeth Zoltan , Cziraki Attila , Szabados Sandor , Horvath Ivan , Koller Akos 
Published: November 2023

BackgroundLocal regulation of blood flow includes extravascular substances released by the parenchymal tissues. Recently, substances derived from fat cells, pericytes and other cells and tissues were shown to exert local vasomotor effects. We hypothesized that pericardial fluid (PF) also affects vasomotor tone, because it has been shown that PF contains vasoactive substances, such as endothelin‐1 (ET‐1).MethodsStudies were performed using isometric myograph (DMT‐Multi Myograph System, pH=7.4, T=36.8 °C). Human PF‐induced changes in vasomotor tone were studied in carotid arteries isolated from Wistar rats. Human PFs was obtained from patients undergoing coronary artery bypass graft (CABG) or valve replacement (VR) surgery and its vasomotor effect was examined by adding PF to the vessels chamber. Vasomotor capacity of vessels was tested by adding KCl (60 mM) before and after the adding PF. Also, in CABG PF the role of ET‐1 was assessed by using the selective endothelin A receptor (ETA) antagonist BQ123 (10−6 M/L).ResultsBoth CABG PF and VR PF induced significant increases in isometric forces of isolated arteries (CABG PF: 3.1±0.7 mN and VR PF 3.0 ± 0.9 mN). Isometric forces induced by CABG PF were significantly reduced in the presence of BQ123 (0.8±0.1 mN vs. 2.6±0.5 mN). We have also observed that contraction to PF were more potent than that of ET‐1. KCl elicited substantial increases in vasomotor tone, both before and after applications of PF (KCl‐1: 6.1±0.2 mN; KCl‐ 2: 6.5±0.9 mN).ConclusionPericardial fluids of patients undergoing cardiac surgery exert substantial constrictor activity in arteries, which in vivo may include microvessels as well. In the vasomotor activity of PF, ET‐1 has a dominant role, but other vasoactive factors may also be present. Thus non‐parenchymal tissues, such as pericardial tissues and fluid may also be involved in the local regulation of coronary blood flow.Support or Funding InformationHungarian Scientific Research Fund (OTKA) K 108444. SROP‐4.2.2.A‐11/1/KONV‐2012‐0024, Hungarian Society of Hypertension 2011/2012, 2013/14.
Stromal Interaction Molecule 1 Disruption Protects Myocardial from Ischemia ‐ Reperfusion Injury
Authors: Mali Vishal R , Ali Maha , Haddox Samuel , Belmadani Souad , Trebak Mohamed , Matrougui Khalid 
Published: November 2023

BackgroundIt has been reported that the increase in Stromal Interacting Molecule 1 (STIM1) expression is associated with myocardial complication. However, the role of vascular STIM1 in cardiac ischemia‐reperfusion (I/R) injury is unknown. Therefore, the central hypothesis of the present study is to determine the role of deleted STIM1 in endothelial cells or smooth muscle cells (SMC) in cardiac I/R injury. Thus, we performed cardiac I/R in control mice, mice lacking the STIM1 specifically in endothelial cells (Stim1−/−EC) or specifically in SMC (Stim1−/−SMC).Methods and ResultsAll mice were subjected to cardiac ischemia for 40 minutes and followed by reperfusion for 24 hours. After 24 hours reperfusion, hearts were incubated with triphenyl tetrazolium chloride to determine the infarct size. In control mice subjected to I/R, the heart develops infarct, which represents 50% of the heart area. Interestingly, in Stim1−/−EC and Stim1−/−SMC, the infarct size was significantly reduced indicating that STIM1 disruption specifically in endothelial cells or SMC protects the heart from I/R injury. Importantly, the heart protection was greater in Stim1−/−SMC than Stim1−/−EC mice. We found an increase in the endoplasmic reticulum (ER) stress in control mice subjected to I/R than in Stim1−/−SMC and Stim1−/−EC mice subjected to I/R.ConclusionOur results indicate that STIM1 disruption in endothelial cells and SMC protects the heart from I/R injury likely through the inhibition of ER stress.Support or Funding InformationNIH
In Utero Exposure to Alcohol Alters Reactivity of Cerebral Arterioles
Authors: Cananzi Sergio Giuseppe , Mayhan William Gerard 
Published: November 2023

BackgroundFetal alcohol spectrum disorders (FASD) have long been linked with developmental abnormalities that often result in brain damage and mental disabilities. It is conceivable that these abnormalities may be related alterations in the regulation of cerebral blood flow (CBF) by in utero exposure to alcohol. Our goal was to examine whether in utero alcohol impaired reactivity of cerebral arterioles during development.MethodsSprague‐Dawley dams were fed a liquid diet with or without alcohol (3% and 6.4%) for the duration of pregnancy (about 21 days). The pups were weaned at three weeks of age. At four weeks of age, we measured CBF (Laser Doppler) in pups in response to hypercapnia. In addition, we made a craniotomy over the parietal cortex and measured responses of cerebral arterioles to agonists that activate endothelial Nitric Oxide Synthase (ADP), neuronal Nitric Oxide Synthase (NMDA), and an agonist that dilates independent of NOS (nitroglycerin). We also used an inhibitor of NOS (L‐NMMA) and tested responses to agonists of eNOS and nNOS. We also measured the expression of NOX‐2 in the brain cortex.ResultsIn utero exposure to alcohol (3 and 6.4%) attenuated rCBF responses to hypercapnia. Responses to agonists that activate eNOS were reduced in cerebral arterioles of alcohol pups. Nitroglycerin produced similar responses in all groups. Protein expression of NOX‐2 from cortex tissue was increased in both 3% and 6.4% alcohol groups.ConclusionsOur findings suggest that in utero alcohol exposure negatively affects the cerebral arterioles and that NOX‐2 may play an important role in the observed impairments. Vascular dysfunction and upregulated expression of NOX‐2 may increase infarct damage from ischemic stroke during FASD.Support or Funding InformationLSU Health Sciences Center‐Shreveport and the Center for Cardiovascular Diseases and Sciences.
Maternal separation stress causes mis‐programming of arterial smooth muscle maturation
Authors: Reho John , Fisher Steven A 
Published: November 2023

Early life stress (ELS) has been implicated in the developmental programming of hypertension however there is little understanding of the programming/mis‐programming of the smaller arteries where vascular resistance and blood pressure are determined. We have previously demonstrated a critical postnatal window for the developmental maturation of the vascular smooth muscle contractile gene program and that this program is under the control of the sympathetic nervous system (SNS). We tested the hypothesis that stress induced hyper‐activation of the SNS during this critical postnatal maturational window would accelerate vascular maturation and potentially lead to adverse cardiovascular effects during adulthood. We used the maternal separation (MS) stress model as a model of ELS where pups were separated from their mothers for 3hrs/day from PND2‐14 and correlated changes in contractile mRNA and resistance mesenteric artery (MA) function throughout postnatal maturation to adulthood. ELS increased expression of CPI‐17, the inhibitory subunit of myosin phosphatase, smMHC, and the a1‐adrenergic receptor, key components of the SNS‐mediated contractile pathway in resistance mesenteric arteries (MA) at PND21 with some effects still present at PND35. This was commensurate with 2–5 fold increases in phenylephrine, KCl, and calcium induced force generation by MAs at PND21. This effect hadt had abated by PND35. Blockade of SNS signaling via the α‐adrenergic blocker Terazosin suppressed this stress induced increases in contractile gene expression and function. No differences were found in adult male rats subjected to ELS under basal or in response to a second stressor. Interestingly, adult females subjected to 1hr restraint stress subsequently demonstrated decreased PE induced constriction of MAs. This was not observed in females subjected to ELS. Thus, ELS results in mis‐programming in the maturation of MA smooth muscle, with evidence for disparate response in males and females. α‐adrenergic receptor blockers administered during ELS may be particularly effective in suppressing the accelerated trajectory of maturation of the constrictor function of these arteries.Support or Funding InformationNIH R01‐HL066171, T32‐HL072751
Exposure to PM2.5 during the First Trimester Contributes to Adult Cardiac Dysfunction
Authors: Tanwar Vineeta , Gorr Matthew , Eichenseer Clayton , Wold Loren 
Published: November 2023

ObjectiveThe present study was designed to investigate the effects of PM2.5 exposure during the first trimester of the in utero period on adult cardiac function.MethodsFemale FVB mice were exposed either to filtered air (FA) or particulate matter (PM2.5) at an average concentration of 51.69 μg/m3 from the Columbus, OH region for 6 h/day, 7 days/wk for the length of the first trimester of pregnancy. To examine cardiac function, male offspring at 12 weeks of age underwent baseline echocardiographic assessment. Following sacrifice, cardiomyocytes were isolated to assess cardiac dysfunction. Protein isolated from LV tissue from separate groups of mice was used to study underlying mechanisms.ResultsMice exposed to PM2.5 during the first trimester demonstrated increased LVESd (2.597 ± 0.082 FA, 2.844 ± 0.077 PM2.5, P=0.04) and LVEDd (3.925 ± 0.062 FA, 4.235 ± 0.084 PM2.5, P=0.01) dimensions and reduced PWTs (1.292 ± 0.039 FA, 1.151 ± 0.039 PM2.5, P=0.02) at 3 months of age. Morphological alterations were associated with lower systolic function as indicated by reduced %FS (34.73 ± 1.741 FA, 29.19 ± 0.817 PM2.5, P=0.007) in PM2.5 exposed mice compared to FA controls. Cardiomyocytes isolated from PM2.5 exposed hearts showed reduced %PS (13.60 ± 0.8970 FA, 10.91 ± 0.7080 PM, P=0.02) compared to FA exposed hearts. −dL/dt was significantly reduced (−4.672 ± 0.3680 FA, 3.581 ± 0.2636 PM2.5, P=0.01) and TR90 (0.1868 ± 0.007295 FA, 0.2158 ± 0.006476 PM, P=0.004) and tau FURA (0.1125 ± 0.008169 FA, 0.1865 ± 0.01068 PM, P=0.0001) were significantly increased between the groups. Western blot analyses demonstrated decreased SERCA‐2 ATPase, p‐PLN and Cav1.2 expression in PM2.5 exposed hearts compared to FA control.ConclusionExposure to PM2.5 during the first trimester of the in utero period triggers adverse developmental changes resulting in global adult cardiac dysfunction similar to exposure during the entire developmental period, suggesting critical windows of priming for adult cardiovascular disease.Support or Funding InformationProject was supported by a grant from the National Institutes of Health (RO1ES019923) to LEW
Nitric oxide donor [Ru(terpy)(bdq)NO]3+ (TERPY) promotes eNOS activation
Authors: Potje Simone Regina , Oliveira Suellen D'Arc , Chen Zhenlong , Bonini Marcelo G , da Silva Roberto S. , Bendhack Lusiane M , Antoniali Cristina , Minshall Richard D 
Published: November 2023

IntroductionTERPY ([Ru(terpy)(bdq)NO]3+) is a ruthenium complex nitric oxide (NO) donor that promotes relaxation of the mesenteric artery and aorta to a greater extent in hypertensive rats as compared to normotensive control. To better understand the mechanism of TERPY action, we sought to investigate whether it acts as both an NO donor and endothelial NO synthase (eNOS/NOS3) activator, as shown previously for nitroglycerin.ObjectiveThe aim of this study was to determine whether TERPY directly or indirectly (i.e., via NOS3 activation) increases the intracellular concentration of NO.MethodsHuman umbilical vein endothelial cells (HUVEC) and human embryonic kidney 293 cells transfected with empty vector (HEK) or eNOS cDNA (HEK‐eNOS) were treated with TERPY (1 μM) for different time lengths (5–60 min). NOS3 expression and Ser1179 phosphorylation were evaluated by Western Blot, and NO accumulation in HUVEC, HEK, and HEK‐eNOS cells was measured using a copper‐based fluorescent probe (1 μM). It was used to quantify the contribution of NO released from the donor source vs NO produced by eNOS (i.e, L‐NAME sensitive NO production).ResultsFirst, we demonstrated that in HEK, TERPY has the ability to release NO without additional stimulus, indicating that this compound is an NO donor. Moreover, in HEK‐eNOS cells, TERPY‐induced NO production was even greater with maximum values observed after 20 min. At this level of NO production, 26% could be blocked by co‐treatment with L‐NAME (non‐selective inhibitor of NOS, 0.1 mM). When cells were stimulated with TERPY for 20 min treatment, TERPY increased NOS3 Ser1179 phosphorylation in HUVEC and HEK‐eNOS further suggesting that TERPY induces NOS3 activation. Consistent with the hypothesis that activated eNOS could be inactivated by Src‐phosphorylated caveolin‐1 as part of a negative feedback mechanism, we have observed caveolin‐1 Tyr14 phosphorylation after 20 min.ConclusionTaken together, these data suggest that TERPY acts as a long‐acting NO donor and also as a NOS3 activator, thereby increasing the NO bioavailability. The potential benefits and mechanisms of eNOS activation by TERPY, as compared to nitroglycerin, requires further characterization in cell cultures and rodent models of hypertension.Support or Funding InformationFinancial support: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) [Grant 232217/2014‐9], Fundação Lemann, and NIH HL‐60678.
N‐myc downstream regulated gene‐1 promotes vascular inflammation through activating both NF‐κB and c‐Jun/AP‐1 pathways in endothelial cells
Authors: Zhang Guanxin , Qin Qing , Yi Bing , Guo Zhifu , Wei Xin , Yin Litian , Yin Yongqiang , Cheng Fang , Sun Jianxin 
Published: November 2023

ObjectiveN‐myc downstream regulated gene‐1(NDRG‐1) is a member of NDRG family that has been shown to play essential roles not only in the regulation of cancer metastasis but also in the development of cardiovascular diseases, such as atherosclerosis and thrombus. The objective of this study is to determine whether NDRG1 is involved in vascular endothelial cell (EC) activation, and if so, to determine the molecular mechanism (s) involved.Methods and ResultsIn response to the stimulation of inflammatory cytokines, such as interleukin‐1β (IL‐1β) and tumor necrosis factor‐alpha (TNF‐α), we show that the expression of NDRG1 in vascular ECs was markedly increased in a dose and time‐dependent manner, as determined by both quantitative real‐time PCR (qRT‐PCR) and western blot analysis. To determine the role of NDRG1 in endothelial activation, we performed a loss‐of‐function study by using NDRG1 specific small interference RNA (siRNA). Our results demonstrate that knockdown of NDRG1 in vascular ECs by siRNA substantially attenuates both IL‐1β and TNF‐α‐induced inflammatory responses in human umbilical vein endothelial cells (HUVECs). The expression of inflammatory cytokines and chemokines, such as IL‐6, IL‐8, and MCP‐1, was markedly inhibited in NDRG1 knockdown ECs. Intriguingly, the IL‐1β‐induced expression of thrombotic markers, such as plasminogen activator inhibitor type 1 (PAI‐1) and tissue factor (TF), was also attenuated in NDRG1 knockdown cells. To further elucidate the functional significance of NDRG1 in vivo, we assessed the expression of NDRG1 in the vessels subjected to the carotid artery ligation‐induced injury. Our results demonstrate that the expression of NDRG1 was markedly increased in the injured vessels as compared with their un‐injured controls. Furthermore, we found that in a mouse model of carotid artery ligation induced injury, knockdown of NDRG1 expression substantially attenuates vascular remodeling, as manifested by decreased neointimal formation, microphage infiltration, and expression of adhesion molecules in injured arteries. Mechanistically, we found that inhibition of NDRG1 markedly inhibited the transcriptional activities of both NF‐κB and AP‐1 pathways in vascular ECs, as determined by both promoter‐driven luciferase and electrophoretic mobility shift assays.ConclusionTaken together, our results for the first time identify NDRG1 as a critical regulator implicated in vascular inflammation and remodeling, and suggest that targeted inhibition of NDRG1 in vasculature may represent a novel therapeutic strategy for preventing certain cardiovascular diseases, such as atherosclerosis and thrombosis.Support or Funding InformationNIH:RO1 HL103869‐01, NSCF: 81541003, SHSCF:15ZR1413400
Enteroendocrine Cell is a Source of VEGF‐A in the Neonatal Intestine
Authors: Yan Xiaocai , Liu Xueli , Managlia Elizabeth , De Plaen Isabelle 
Published: November 2023

Necrotizing enterocolitis (NEC) is the most common gastrointestinal disease affecting premature newborns, characterized with a high mortality and chronic morbidity. We have previously found that VEGF‐A is decreased in human and experimental NEC and may play a role in NEC. However, which cell type(s) produce(s) VEGF‐A in the neonatal intestine remains unknown. Here, to determine the cellular origin of VEGF expression in normal mouse and human intestines, fixed small intestinal tissues of newborn mice and a human neonate with intestinal atresia were examined by immunofluorescence staining for VEGF‐A together with CD31, chromogranin A. To determine whether VEGF‐A was expressed in monocytes, intestines from CX3CR1‐GFP mice were used. In both intestinal tissues of mouse and human neonates, we found that VEGF‐A is strongly present in epithelial cells expressing chromogranin A, in myenteric neural plexus cells and in CD31+ cells. However, VEGF‐A was not detected in CX3CR1+ cells in the mouse intestine or in CD45+ cells in the intestine of humans. In conclusion, VEGF‐A is strongly expressed in enteroendocrine cells, myenteric neural plexus cells and endothelial cells, but not in myeloid‐derived cells. Therefore, we speculate that enteroendocrine cells may be critical for supporting the development of the intestinal villi microvasculature.Support or Funding InformationMemorial Foundation (Friends of Prentice) and NIH grant R01HD060876 (IDP).
Heme Oxygenase‐1 Counteracts HIV Protease Inhibitor‐Mediated Endothelial Cell Dysfunction
Authors: Durante William , Liu Xiao‐ming , Durante Zane E. , Peyton Kelly J. 
Published: November 2023

The use of human immunodeficiency virus (HIV) protease inhibitors has extended the duration and quality of life for HIV‐positive individuals. However, there is increasing concern that this antiviral therapy may promote premature cardiovascular disease by impairing endothelial cell (EC) function. In the present study, we evaluated the effect of HIV protease inhibitors on EC function and determined if the vasoprotective enzyme heme oxygenase‐1 (HO‐1) influences the biological action of these drugs. Treatment of human umbilical vein ECs with three distinct protease inhibitors, including ritonavir, atazanavir, and lopinavir, stimulated the expression of HO‐1 protein in a time‐ and concentration‐dependent manner. The induction of HO‐1 protein was preceded by a rise in HO‐1 mRNA and resulted in a significant increase in HO activity. Furthermore, protease inhibitors evoked a marked increase in Nrf2 activity and an elevation in the production of reactive oxygen species (ROS). Protease inhibitors also stimulated HO‐1 promoter activity, and this was prevented by mutating the antioxidant responsive elements of the promoter or by overexpressing a dominant‐negative mutant of Nrf2 that lacked the transactivation domain. In addition, the protease inhibitor‐mediated induction of HO‐1 and activation of Nrf2 was abolished by the antioxidant N‐acetyl‐L‐cysteine. Finally, protease inhibitors blocked EC proliferation and DNA synthesis in a concentration‐dependent fashion that was potentiated by the HO‐1 inhibitor tin protoporphyrin‐IX or by silencing the expression of HO‐1. In conclusion, this study found that HIV protease inhibitors stimulate HO‐1 gene expression in ECs via the ROS‐Nrf2 pathway, and that the induction of HO‐1 counteracts the anti‐proliferative action of protease inhibitors. Therapeutic strategies targeting HO‐1 represent a novel approach in alleviating protease inhibitor‐mediated EC dysfunction and vascular disease in HIV‐positive patients.Support or Funding InformationSupported by NIH grant R01 HL59976.