bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Anti‐Inflammatory Cytokine IL‐19 Reduces mRNA‐Stability Protein HuR Function in Human Vascular Smooth Muscle Cells
Authors: Herman Allison , Haines Dale , Autieri Michael V. 
Published: November 2023

Vascular diseases such as atherosclerosis and restenosis are often exacerbated by associated inflammation. Vascular smooth muscle cells (VSMCs) respond to injury and are capable of producing pro‐inflammatory cytokines which amplify the inflammatory response. Identification of compounds which attenuate this response is an important therapeutic goal. Presently, little is known about the potential for endogenously expressed anti‐inflammatory cytokines to attenuate inflammation and have vascular protective effects. Interleuken‐19 (IL‐19) is a newly identified member of the IL‐10 anti‐inflammatory family. IL‐19 expression can be induced in VSMCs by pro‐inflammatory cytokines and is capable of reducing the stability and abundance of pro‐inflammatory mRNA transcripts. HuR is an mRNA stability protein which functions to stabilize pro‐inflammatory transcripts by shuttling from the nucleus to the cytoplasm where it binds to AU‐rich elements within the 3′ UTR of transcripts. Nuclear to cytoplasmic shuttling is essential for HuR activity. We previously determined that treatment of VSMCs with IL‐19 transiently reduces HuR abundance in the cytoplasm, by a presently unknown mechanism. In the present study we show that overexpression of HuR significantly increases VSMC proliferation. We hypothesize that IL‐19 inhibits HuR translocation to the cytoplasm by altering chaperone protein function, abundance, or both. To test this hypothesis, VSMCs were transfected with epitope‐tagged HuR expression plasmids, followed by treatment with TNFα alone, IL‐19, or both, to determine if any HuR‐interacting proteins are differentially regulated by inflammation or IL‐19 to alter HuR activity. Protein extracts were prepared from each treatment group and HuR was immunoprecipitated and proteomic analysis (MASS SPEC) was performed to identify HuR‐interacting proteins in each of these conditions. Numerous novel protein interactions with HuR are altered by the different inflammatory treatment conditions. The roles of many of these novel interacting proteins involve RNA‐binding and post‐transcriptional regulation based on the literature. Inflammation‐modulated differential HuR protein‐ protein interactions provide a framework showing mRNA stability can be modified by HuR chaperone proteins VSMCs, which is a novel direction of study for inflammation. The proteins identified could lead to a better understanding of the mechanisms associated with vascular inflammatory diseases such as atherosclerosis and provide targets for new therapeutic agents.Support or Funding InformationThis work was supported by grants HL115575 and HL117724 from the National Heart Lung, and Blood Institute of the National Institutes of Health, and Grant 13GRNT1685003 from the American Heart Association to MVA
The role of PKC‐B in the BKCa functionality of vascular smooth muscle of diabetic rats
Authors: Espinosa‐Tanguma Ricardo , Álvarez‐Méndez Juan Antonio , Hernández‐García Ana Patricia , Algara‐Suárez Paola 
Published: November 2023

Diabetes mellitus (DM) is a disease characterized by high blood glucose concentrations. Cardiovascular disorders associated with DM are the leading cause of high morbidity and mortality of this disease, where endothelial tissue as well as smooth muscle is damaged. For example, the contractility of the smooth muscle cells is increased partly due to a decrease in the activity of the calcium‐dependent high conductance potassium channels (BKCa), which along with other potassium conductance maintain membrane potential and vascular tone.Protein kinase C (PKC) is involved in signaling pathways that promote muscle contraction. Christiandimitropoulou (2002) reported that in the vascular smooth muscle of diabetic rats the outwardly currents flowing through the BKCa channels are decreased. Likewise it was also reported that PKC can phosphorylate some residues of the carboxyl terminal of the alpha subunit of said channel. Data from our laboratories show that the inhibition of the classic isoforms of the PKC is associated with a decrease in the contraction of the mesenteric artery of diabetic rats, which suggests that the beta isoform may be involved in the vascular damage observed in DM.ObjectivesTo study the role of the PKC‐B on BKCa channels functionality and the aortic vascular contractility of diabetic rats induced by streptozotocine.MethodsTension and western blot experiments were performed on aortic rings.Resultsthe NS11021, a BKCa channel activator, significantly relaxes the rings of diabetic rats on a lesser scale than those of the control group, the PKC‐B LY333531 inhibitor does not modify the contractility induced by phenylephrine in normal rings but importantly relaxes those from diabetic rings. The effect of NS11021 is preserved even in the presence of LY333531. Western blot experiments suggest that smooth muscle from diabetic rats is less phosphorylated, indicating that BKCa activity is reduced compared with those from control rats.Conclusionsvascular smooth muscle of diabetic rats show greater contractility and this may be due to a diminished activity of the BKCa channels.Support or Funding InformationSupported by C14‐FAI‐04‐33.33 to PAS, Conacyt 223350‐M to RET. Conacyt scholarship to JAM (295659)
Clustering and combining pattern of metabolic syndrome risk factors and its association with arterial stiffness
Authors: Vicente Wanda Rafaela , Rodrigues Sara , Dutra‐Marques Akothirene CB , Carvalho Jefferson C , Jordao Camila P , Toschi‐Dias Edgar , Hussid Maria Fernanda , Costa‐Hong Valeria , Bortolotto Luiz A , Cepeda Felipe X , Trombetta Ivani Credidio 
Published: November 2023

IntroductionMetabolic syndrome (MetS) is associated with structural and functional vascular abnormalities that can lead to increased arterial stiffness, which predicts cardiovascular events and increases mortality. However, little is known regarding the implications of risk factor clustering in the arterial stiffness.ObjectiveIn MetS, determine the clustering and the combining pattern of risk factors and its association with arterial stiffness.MethodsWe selected recently diagnosed patients with MetS (n=47, 51 ± 6 y) according to ATP III criteria. A control group (C, n=17, 50 ± 4 years) was also studied. Arterial stiffness was evaluated by pulse wave velocity (PWV) in carotid‐femoral segment. The patients were categorized and analyzed according MetS risk factors clustering (3, 4 and 5 factors). We performed correlation tests and multiple linear regressions to determine predictor variables with PWV and independent variables of sex, age, body mass index, and MetS risk factors.ResultsPatients with MetS had increased PWV compared to C (8.8 ± 0.7, 9.8 ± 1.4 m/s, p=0.01). In multivariate analysis, the variables that remained as predictor of PWV were age (β=0.35, p= 0.01) and triglycerides (β=0.29, p= 0.03). There was a progressive increase of PWV to increase the number of risk factors. Furthermore, the PWV compared to C group was similar for cluster of 3 risk factors, whereas were higher for cluster of 4 and 5 risk factors (8.7 m/s [7.8 to 10.0], 9.4 m/s [7.7to 11.7], 9.8 m/s [7.3 to 13.4] and 10 m/s [9.3 to 13.4]).ConclusionThe number of risk factors amplifies the arterial stiffness. The presence of alteration in the triglycerides worsening the stiffness of large vessels, emphasizing the importance in addressing this risk factor for MetS patients.Support or Funding InformationThis study was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP# 2011/17533‐6 ) and, in part, by Fundação Zerbini. Sara Rodrigues was supported by FAPESP# 2013/15323‐0, Edgar Toschi‐Dias was supported by FAPESP# 2013/07651‐7, Felipe X Cepeda was supported by FAPESP# 2015/03274‐0.
Hypoxic Regulation of Vascular Proteasomal Degradation of MLCK
Authors: Adeoye Olayemi Olufikayo , Sorensen Dane William , Williams James , Pearce William 
Published: November 2023

Chronic exposure of in‐utero fetuses to severe hypoxia induces maladaptive changes in vascular structure and function, and elevates postnatal susceptibility to metabolic, endocrine, cardiovascular diseases. Correspondingly, multiple studies have demonstrated that chronic hypoxia depresses vascular contractility through mechanisms that remain to be fully elucidated. Our recent studies have shown that in fetal arteries chronic hypoxia decreases the abundance and activity of myosin light chain kinase, which is the rate‐limiting enzyme for force generation in smooth muscle cells. Here we hypothesize that chronic hypoxia decreases vascular contractility via age dependent increases in proteasomal degradation of MLCK protein. To test this hypothesis we used a well established chronic hypoxia model involving pregnant ewes maintained at sea level or at altitude (3280m) during the last 110 days of gestation. In vitro contractility experiments revealed that chronic hypoxia decreased maximum myogenic stress by 56%. Chronic hypoxia also significantly decreased MLCK abundance, and moreso in fetal (90%) than adult (47%) arteries. Conversely, qPCR measurements of MLCK mRNA revealed similar abundances in adult and fetal arteries, suggesting that increased loss of MLCK was not due to inhibition of transcription. To more directly elucidate mechanisms of MLCK degradation, the effects of MG132 (a potent inhibitor of proteasomal degradation) were quantified organ cultured fetal and adult arteries. Rates of MLCK degradation averaged 0.83 ± 0.21 in fetal and 0.99 ± 0.38 in adult normoxic arteries. With chronic hypoxia, degradation rates increased by 53% and 61% in fetal and adult arteries respectively. Together these data suggest that depressed contractile activity during hypoxic acclimatization occurs via hypoxic promotion of proteasomal degradation of MLCK slightly more in adult than fetal arteries.Support or Funding InformationThese studies were supported by an NIH Program Project grant from NICHD (HD31266,) and an NIH R01 grant from NINDS (NS076945).
Passive Heat Therapy as a Novel Approach for Inducing Angiogenesis in Humans: Roles of Nitric Oxide
Authors: Brunt Vienna E , Needham Karen Wiedenfeld , Comrada Lindan N , Francisco Michael A , Minson Christopher T 
Published: November 2023

Chronic passive heat therapy is a novel health intervention which may have widespread cardiovascular benefits. Studies in animals suggest chronic heat exposure promotes angiogenesis via NO, independently of VEGF. We sought to determine whether the same occurs in humans using an endothelial tubule formation assay to quantify serum angiogenic balance.METHODSSerum was collected from ten young (23±2 years), healthy human subjects before and after 8 weeks of passive heat therapy (36 sessions of hot water immersion sufficient to maintain rectal temperature ≥38.5°C for 60min per session; post‐heat therapy serum was collected at least 36h after the last session). All analyses were performed in duplicate using human sera collected pre and post heat therapy. To assess serum angiogenic balance, human umbilical vein endothelial cells (HUVECs) were plated onto serum‐free Matrigel, exposed to human sera, and incubated for 10h, after which total tubule length per frame was determined by two blinded investigators. To investigate whether potential improvements in serum angiogenic balance following heat therapy were NOS‐dependent, angiogenesis experiments were repeated in which HUVECs were additionally treated with 1mM L‐NNA, a competitive inhibitor of NOS. In separate experiments, HUVECs were exposed to human sera for 24h and eNOS protein was determined using Western blot, normalized to vinculin loading control. Lastly, serum VEGF concentration was determined by ELISA.RESULTSSerum angiogenic balance was improved following 8 weeks of heat therapy (total tubule length: 71.8±1.0 to 74.3±1.1mm, p=0.03) and this improvement was prevented by L‐NNA (73.1±1.7 to 73.9±1.1, p=0.56). eNOS protein expression increased by 18.6±3.4% (p=0.05) in endothelial cells exposed to human sera following heat therapy relative to pre‐heat therapy. Heat therapy decreased serum VEGF concentration (370±77 to 331±69 pg/ml, p=0.05).CONCLUSIONSChronic passive heat therapy in humans upregulates circulating factor(s) which promote angiogenesis in a NOS‐dependent and VEGF‐independent manner, possibly through increasing eNOS expression. These results are consistent with those reported in animal studies and suggest chronic passive heat therapy could be a viable treatment option for improving vascularization in at‐risk populations.Support or Funding InformationSupported by AHA Grant #14PRE20380300 and the Eugene and Clarissa Evonuk Memorial Foundation
DPPI Deficiency Enhances Both Angiogenesis and Arteriogenesis and Improves Cardiac Function after Myocardial Infarction
Authors: Kolpakov Mikhail A , Hooshdaran Bahman , Guo Xinji , Wang Tao , Rafiq Khadija , Vlasenko Liudmila , Qi Zhao , Garikipati Venkata NS , Kishore Raj , Houser Steven R , Sabri Abdelkarim 
Published: November 2023

Study objectiveMyocardial infarction (MI) induces both hypoxia and inflammation that trigger angiogenesis. Numerous proangiogenic and proarteriogenic factors are released during the inflammatory reaction post‐MI. However, the overall effect of the various factors of the inflammatory component depends on the local environment. This study examined the potential role of inflammatory serine protease (ISP) inhibition on angiogenesis after MI.Methods and resultsWe subjected wild‐type (WT) and mice lacking Dipeptidyl peptidase I (DPPI), a lysosomal enzyme involved in the cleavage and activation of major ISPs, to MI up to 4 weeks and assessed the role of DPPI gene deletion on angiogenesis and cardiac remodeling and function.DPPI deficient mice show markedly reduced activity of neutrophil‐ and mast cell‐derived serine proteases after MI compared to WT mice, along with a reduction in infarct size and improved cardiac function. DPPI deficiency also increased the number of capillaries and mature vessels in infarcted hearts by upregulating the expression of angiogenic cytokines such as vascular endothelial growth factor (VEGF) and angiopoietin‐1. Investigation of the mechanisms involved show reduced levels of soluble VEGF receptor 1 (sVEGFR1) accumulation in DPPI KO infarct together with increased phosphorylation of VEGFR2. The negative role of inflammatory serine proteases on angiogenesis was further demonstrated in vitro in tube formation assay. Treatment of human umbilical vein endothelial cells with the neutrophil‐derived serine protease cathepsin G (CG) led to up‐regulation of sVEGFR1 and its interaction with VEGFA, decreased VEGFR2 signaling and reduced the number and length of endothelial cell tubes. In contrast, preincubation of endothelial cells with neutralizing anti‐sVEGFR1 antibodies attenuated endothelial cell tube disorganization, improved VEGFR2 signaling and preserved new vessels formation.ConclusionsOur findings reveal the role of ISPs as key components of a molecular mechanism that negatively modulates angiogenesis through regulation of the relative levels of sVEGFR1 in endothelial cells. Our study also point to DPPI inhibition as a potential therapeutic target for stimulation of angiogenesis and the maintenance of cardiac function post‐MI.Support or Funding InformationThis work was supported by the National Institute of Health (HL088626 and HL076799) and American Heart Association (15SDG22650004 and 14PRE20380518).
Angiotensin‐(1–7) Requires both Mas1 and AT1 Receptors to Restore Angiogenesis
Authors: Stodola Timothy , Exner Eric , Hoffmann Brian , Didier Daniela , Greene Andrew 
Published: November 2023

The Renin‐Angiotensin System plays an important role in many physiological processes such as sodium homeostatis, regulating vascular tone and angiogenesis. The octapeptide Angiotensin II (AngII) acting through the AT1 receptor has long been known to promote vasoconstriction, fibrosis, cell proliferation and skeletal muscle angiogenesis in response to exercise or electrical stimulation. A second vasoactive peptide of the Renin‐Angiotensin System, Angiotensin‐1–7 (Ang‐(1–7)), acts through the Mas1 receptor and is generally thought to have many physiological actions that are antagonistic to the actions of the AngII‐AT1 axis, such as vasodilation, anti‐fibrotic and anti‐proliferative effects. We hypothesized Ang‐(1–7) acting through the Mas1 receptor could restore skeletal muscle angiogenesis in response to electrical stimulation. Little is known about the signaling events initiated by Mas1 receptor activation by Ang‐(1–7) relative to AngII‐AT1 activity. Here we show in Sprague Dawley rats with Renin suppressed through high dietary salt intake, which suppresses angiogenesis, that infused Ang‐(1–7) acting through the Mas1 receptor restores angiogenesis in vivo in skeletal muscle in response to electrical stimulation and in vitro in a tube formation assay using rat microvascular endothelial cells, in both cases independent of AngII. Using advanced proteomic methodology to characterize the components of Mas1 receptor signaling we identified signaling pathways that may play a role in the observed angiogenic response, such as nascent protein synthesis, exocytosis, cellular reorganization and regulators such as NO, NF‐kappaB and ERK1/2 signaling. Inhibition of ERK1/2 signaling inhibited tube formation by endothelial cells stimulated by AngII or Ang‐(1–7). We further assessed angiogenesis in response to electrical stimulation promoted by Ang‐(1–7) in SS‐Mas1em1Mcwi (Mas1 mutant knockout) and SS‐Agtr1aem5Mcwi (AT1 mutant knock out) strains, and compared to wild‐type SS/Mcwi. We found reduced angiogenesis to electrical stimulation after Ang‐(1–7) or orally active Ang‐(1–7) analog AVE0991 treatment in both the Mas1 mutant and the AT1 mutant knock outs relative to the SS/Mcwi. The RAS is a complex physiological system that can affect a wide range of processes in the cardiovascular system. Even though the AT1 and Mas1 receptor axes largely have antagonistic functions, we have shown that capillary angiogenesis can both be restored by AngII‐AT1 or Ang‐(1–7)‐Mas1 activity, and the restoration depends on the presence of functioning Mas1 and AT1 receptors.Support or Funding InformationNIH Heart, Lung and Blood Institute 082798
Hypoxia and Ischemia Regulate Placental Syndecan Expression and Shedding
Authors: George Eric M 
Published: November 2023

Preeclampsia, a hypertensive disorder of gestation, affects ~5% of the population, and is a leading cause of fetal and maternal morbidity and mortality. While the origins of the disease are unclear, it is commonly accepted that placental ischemia/hypoxia is a central causative factor. In response, the placenta secretes soluble factors which cause the maternal manifestation of the disorder. Among the pathways believed to be important in this response is activation of innate immune mechanisms, such as inflammatory cytokines. It has been widely reported in sepsis that vascular inflammation induces remodeling and shedding of the vascular endothelial glycocalyx, in particular cleavage and shedding of members of the integral membrane syndecan family. It is now known that these shed fragments have biological activity and act as potent inducers of the innate immune and inflammatory response. What is less well known is the presence of a functional glycocalyx in the placenta. Here we hypothesize that hypoxia will induce changes in placental syndecan production and shedding in vitro, and that chronic placental ischemia will cause syndecan shedding from the placenta in vivo. To examine the effects of hypoxia on placental cells, we cultured BeWo cytotrophoblast cells in oxygen tensions mimicking healthy (8%) and ischemic (1%) placentas, and examined syndecan1–4 mRNA and soluble SDC1 in the media. In response to hypoxia, SDC1,2, and 4 mRNA increased significantly (14, 6.5, and 2.5 Fold, respectively, p<0.05 each), as assayed by TaqMan qRT‐PCR. SDC3, in contrast, exhibited no significant difference in expression. Soluble SDC1 in the media, meanwhile was significantly increased by ~50% compared to normoxic control as measured by ELISA (41 ± 4 vs 63 ± 4 pg/ml, p<0.05). To determine the effects of chronic placental hypoxia/ischemia on SDC, we utilized the reduced uterine perfusion pressure (RUPP) rodent model. RUPP rats exhibited significantly increased blood pressure on gestational day 19 when compared to their control counterparts (105 ± 4 vs 122 ± 6 mmHg, p<0.05). Interestingly, soluble circulating SDC1 in the maternal circulation was significantly increased in RUPP rats when compared to their control counterparts (11 ± 4 vs 99 ± 20 pg/ml, p<0.05). These data suggest that hypoxia/ischemia differentially regulate SDC isoforms in vitro. Furthermore it is possible that isoform specific or pan‐syndecan circulating levels of syndecans could serve as a biomarker for preeclampsia. The importance of syndecan fragments in the inflammatory cascade during preeclampsia remains to be determined, as do the mechanisms regulating syndecan production and cleavage in these tissues.Support or Funding InformationThis work was supported in part by P01HL51971, P20GM104357 (EMG), and R00HL116774 (EMG).
Angiogenic Imbalance and Hypertension during Pregnancy Results in Intrauterine Growth Restriction and Developmental Delays in Rodent Offspring
Authors: Bowles Teylor , Golden Leia , Spencer Shauna‐Kay , Owens Michelle Y , Wallace Kedra 
Published: November 2023

IntroductionIn the current study our objective was to determine if rat pups born to dams experiencing angiogenic imbalance and hypertension during pregnancy had intrauterine growth restriction (IUGR) and developmental delays. To test this objective we infused sFlt‐1 and sEndoglin into timed pregnant rats. This animal model creates pathological symptoms similar to what is seen in women with Hemolysis Elevated Liver enzyme Low Platelet (HELLP) syndrome and severe preeclampsia (sPreE). Additionally these pregnant rats development hypertension and have increased production of TNF‐alpha, IL‐6 and IL‐17 which is also experienced by women with HELLP and sPreE.MethodsOn gestational day (GD) 12, mini osmotic pumps infusing sFlt‐1 and sEndoglin (4.7 and 7 ug/kg respectively) were implanted into timed pregnant rats (n=4) to create HELLP or sPreE. Within 12hrs of delivery 4 pups from each litter of HELLP rats and normal pregnant (NP; n=4) rats were cross‐fostered with a set of lactating SHAM rats (n=4) so that the final litter size per SHAM rat was 16 pups. Rat pups underwent the standard Irwin battery of developmental milestones beginning on post‐natal day (PND) 2 – PND16.ResultsThere was not a difference in the gestational day of delivery between the groups, but pups in the HELLP group did display IUGR compared to pups in the NP group (6.16 ± 0.10 vs. 6.69 ± 0.13g in NP; p=0.004). There was a significant interaction (p<0.0001) in surface righting with pups displaying the largest delay in this task at PND2; 35.06 ± 9sec in HELLP rats vs. 18.41 ± 3.73sec in NP rats. Significant interactions between the days of testing were seen in negative geotaxis (p<0.0001), cliff avoidance (p<0.0001) and open circle (p<0.0001). Rats in the HELLP group had a significant delay in achieving the forelimb grasp test compared to NP rats (p<0.0001), with up to 39% of the NP having achieved this task by PND5 compared to only 15% of the HELLP group. Pups born to dams with HELLP had a significant delay in eye opening compared to NP pups (p=0.0043).ConclusionsThese data support the hypothesis that IUGR is associated with developmental delay. The angiogenic imbalance during pregnancy in this model may serve as one mechanism whereby IUGR occurs in the absence of pre‐term delivery, thereby mediating the developmental delay in the resulting offspringSupport or Funding InformationThis work was funded by an Institutional Development Award (IDeA) from the NIGMS under grant number P30GM103328.