bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comorbidity Between Metabolic Syndrome and Depression Exacerbates Oxidative Stress and Inflammation in the Brain
Authors: Branyan Kayla , DeVallance Evan , Asano Shinichi , Lemaster Kent , Allen Colton , Perry Sydney , Frisbee Jefferson , Chantler Paul 
Published: November 2023

Major cardiovascular disease risk factors, such as metabolic syndrome (MetS) and depression, are associated with a chronic inflammatory state and increased oxidative stress. As in other peripheral microvessels, inflammation and oxidative stress impair the microvasculature of the brain, leading to increased risk of stroke. It is unclear how the comorbidity between MetS and depression affect the inflammatory and oxidative stress conditions in the brain. To determine if depression and MetS together further exacerbate cerebral inflammation and oxidative stress compared to either disease alone, markers of oxidative stress were assessed through dihydroethidium (DHE) and DAF‐FM assays along with PCR and ELISA to evaluate the inflammatory response in the brains of obese Zucker rats (OZRs), a model of MetS, and lean Zucker rats (LZRs), healthy controls. Reactivity of the middle cerebral artery (MCA) was used as a measure of vascular function. The MCA was isolated and then hung in a pressurized vessel myobath to be exposed to increasing doses of acetylcholine (Ach) to test endothelium‐dependent dilation. OZRs had increased superoxide production in the basilar arteriole (p<0.05), which was exacerbated with UCMS. Similarly, the OZRs and OZR‐UCMS had a significant decrease in nitric oxide production as compared to both LZR groups. Concentrations of anti‐inflammatory cytokines, IL‐4 and IL‐10, were reduced in OZRs and further reduced in OZR‐UCMS. Inflammatory marker (TNFα and CD68) mRNA abundance was elevated in OZRs and to a higher extent in OZR‐UCMS. This oxidative stress and inflammation contributed to decreased MCA reactivity in OZR controls from that of LZR controls in response to Ach (p<0.01). In both LZRs and OZRs, MCA dilation was further attenuated with UCMS as compared to their respective control values. These data show that the comorbidity of MetS and depression exacerbates the oxidative stress and inflammatory states associated with each disease separately. Our results suggest that the cerebral environment is drastically changed during this comorbidity and leads to vascular dysfunction, which can cause an increased risk of poor cardiovascular outcomes.Support or Funding InformationNational Institute of Health (5P20GM109098)
Amyloidogenic Medin Induces Endothelial Dysfunction and Vascular Inflammation Through the Receptor for Advanced Glycation Endproducts
Authors: Migrino Raymond Q. , Davies Hannah , Truran Seth , Karamanova Nina , Franco Daniel A. , Serrano Geidy , Beach Thomas , Madine Jillian 
Published: November 2023

AimsMedin is a common amyloidogenic protein in humans, especially in older individuals. It has been implicated in the development of arterial degenerative changes associated with vascular aging. The mechanisms underlying medin‐induced vascular injury remain unknown. The aims are to assess medin's effects on human peripheral and central arteriole endothelial function, nitrative stress and vascular inflammation and identify potential mechanisms of injury.Methods and ResultsEx‐vivo human adipose and leptomeningeal arterioles were exposed (1 hour) to medin (0.1, 1 or 5 μM) without or with FPS‐ZM1 (100 μM, receptor for advanced endglycation products [RAGE]‐specific inhibitor) and endothelial function (acetylcholine dilator response) was compared to baseline control. Medin's effect on adipose arteriole smooth muscle function was tested by comparing post‐exposure dilator response to DETA NONOate (a nitric oxide [NO] donor) compared to baseline response. Human umbilical vein endothelial cells were exposed to medin (20 hours) without or with FPS‐ZM1 and oxidative and nitrative stress, cell viability and pro‐inflammatory signaling measures were obtained. Medin caused impaired endothelial function (versus baseline response to 10−4 M acetylcholine: −38.6±6.7% and −35.0±8.9% in adipose and leptomeningeal arterioles, respectively, each p<0.05) in human adipose and leptomeningeal arterioles. Medin did not affect adipose arteriole dilator response to DETA NONOate. Medin‐induced endothelial dysfunction was reversed by RAGE inhibitor. Medin decreased endothelial cell NO production and cell viability, increased peroxynitrite, superoxide production and profoundly increased gene and protein expression of interleukin‐6 and interleukin‐8 via activation of nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NFκB). These effects were reversed by RAGE inhibitor.ConclusionsMedin causes human microvascular endothelial dysfunction, oxidative and nitrative stress and pro‐inflammatory signaling mediated via RAGE. The discovery of this novel mechanism of injury may have implications in our understanding of and development of treatments to reverse vascular aging.Support or Funding InformationThe work was supported by National Institutes Health (NIA R21AG044723, NINDS U24NS072026, NIA P30AG19610, NIA RO1AG019795); Veterans Affairs Merit grant (I01BX007080); British Heart Foundation (FS/12/61/29877); the Arizona Department of Health Services (contrast 211002); the Arizona Biomedical Research Commission (4001, 0011, 05‐901 and 1001); Michael J. Fox Foundation for Parkinson's Research and Amyloidosis Foundation. The study was based on work supported by the Department of Veterans Affairs and VA employment.
Dynamics of mitochondrial function changes in the cerebral vasculature of type 2 diabetic rats
Authors: Merdzo Ivan , Rutkai Ibolya , Sure Venkata N. L. R. , McNulty Catherine A. , Katakam Prasad V. , Busija David W. 
Published: November 2023

Mitochondrial dysfunction has been suggested as a potential underlying cause of pathological conditions associated with type 2 diabetes (T2DM). We have previously shown that mitochondrial respiration levels assessed by the Seahorse XFe24 analyzer in large cerebral arteries, and mitochondrial protein levels measured by western blot in cerebral arteries and microvessels, were similar in insulin resistant, non‐diabetic, Zucker obese rats and lean control rats (Am J Physiol 310:H830‐838, 2016). In the present study we extended our investigations into mitochondrial dynamics and superoxide production of the cerebral vasculature of 14 week old Zucker diabetic fatty obese rats (ZDFO) with early T2DM.Body weight and blood glucose levels were significantly higher in the ZDFO compared to the control, lean (ZDFL) rats (347.2 ± 10.18 g vs. 320.4 ± 3.9 g, n = 20, P < 0.05; and 26.33 ± 1.56 mmol/L vs. 9.18 ± 0.46 mmol/L, n = 20, P < 0.05; respectively). In addition, basal mitochondrial respiration and proton leak were significantly decreased in the large cerebral arteries of the ZDFO rats compared with ZDFL (181.4 ± 10.93 pM/min/μg protein vs. 129.7 ± 8.211 pM/min/μg protein, n = 59, P < 0.05; and 149.3 ± 10.41 pM/min/μg protein vs. 100.3 ± 7.56 pM/min/μg protein, n = 59, P < 0.05; respectively). Furthermore, superoxide production was significantly increased in the microvessels of the ZDFO group (2355 ± 628 vs. 208 ± 48, n = 10, P < 0.05). The expression of mitochondrial proteins total MnSOD and VDAC were significantly lower in the cerebral microvessels (90.35 ± 9.13% vs. 123.1 ± 12.77%, n = 13, P < 0.05; and 23.63 ± 2.62% vs. 30.54 ± 2.07%, n = 12, P < 0.05; respectively), and levels the acetylated or active form of MnSOD were significantly reduced in large arteries of ZDFO group (89.72 ± 3.17% vs. 52.23 ± 6.01%, n = 12, P < 0.05). Despite evidence of increased oxidative stress in ZDFO, exogenous superoxide dismutase was not able to restore mitochondrial respiration in ZDFO rats.Our results show for the first time that mitochondria of large arteries and microvessels are adversely affected in the cerebral vasculature during the early stages of T2DM.Support or Funding InformationThis work was supported by National Institutes of Health grants (DWB: HL‐077731and HL093554), Louisiana Board of Regents Support Fund‐Research Competitiveness Subprogram (PVK: LEQSF(2014‐17)‐RD‐A‐11), American Heart Association National Center NRCP Scientist Development Grant (PVK: 14SDG20490359), and American Heart Association Post‐Doctoral Fellowship Grant (IR: 15POST23040005). This research was also supported by the Louisiana Board of Regents Endowed Chairs for Eminent Scholars program (DWB).
Can exercise protect against chronic stress induced cerebral microvessel changes in Lean and Obese Zucker rats?
Authors: Sheets Whitney Jordan , Asano Shinichi , Dawson Mariah , Ashman Melissa , Clayton Stuart , Brooks Steven , Frisbee Jefferson , Chantler Paul 
Published: November 2023

BackgroundChronic stress‐induced depression is a major cardiovascular risk factor, and metabolic Syndrome (MetS) is linked with depression. The relationship among chronic stress/depression (UCMS), MetS, CVD, and cerebrovascular dysfunction is poorly understood. We determined how UCMS and Exercise training affected cerebral microvessels density (MVD), and the pro (nitric oxide, VEGF, STAT3) and anti (TSP‐1) angiogenic factors. We hypothesize that UCMS will decrease MVD in Obese Zucker Rats (OZR = MetS), and exercise training will prevent this rarefaction by increasing pro‐angiogenic mediators, while reducing anti‐angiogenic factors including oxidative stress.MethodsNine‐week old lean rats (LZR) and OZR were randomly assigned (n=6/group) to: Control(C), Exercise (EX), subjected daily to an UCMS, and EX+UCMS for 8 weeks. At 17 weeks, brains were removed and snap frozen in Tissue Tek medium. Cortex MVD were measured by fluorescence IHC (rat anti‐CD31 antibody). Oxidative stress was assessed in cerebral tissue by ELISA technique (anti 3‐Nitrotyrosine antibody) and by DHE. Cerebral inflammatory and angiogenic markers for STAT3, VEGF and TSP‐1 were measured by western blot or ELISA.ResultsMVD was decreased (−31%, p<0.05) in the cortex from OZR‐UCMS vs. OZR control. Exercise training attenuated UCMS induced decreased MVD in OZR by 41%. In contrast, there were no significant differences (8% decrease to 4% increase) in MVD among LZR groups. In both LZR and OZR UCMS increased oxidative stress, and decreased nitric oxide in the cerebral vessels. Although VEGF results were not yet significant, a slight decrease was seen in OZR UCMS that was recovered with exercise training in UCMS. Initial data also suggests that TSP‐1, which is higher in OZR vs. LZR control, is also influenced by UCMS, and Ex. Compared to LZR‐C, LZR‐UCMS had an 74% increase in STAT3. Although both OZR‐C and OZR UCMS exhibited significantly increased STAT3 phosphorylation compared to LZR‐C, there was no difference between OZR‐C and OZR UCMS. Both LZR‐Ex and OZR‐Ex attenuated UCMS induced elevated STAT3 activity.ConclusionTaken together, UCMS drives the development of decreased brain MVD in OZR. Chronic stress and obesity both independently increased STAT3 signaling in the brain, which was then abrogated by exercise in both lean and obese animals. Exercise also prevented the onset of cortical rarefaction seen in OZR‐UCMS. We are further exploring TSP1 and VGEF pathways.Support or Funding InformationAcknowledgements: American Physiological Society Short‐Term Experience for Underrepresented Persons Fellowship and the National Institute of Diabetes and Digestive and Kidney Diseases for helping to fund this project. National Institute of Health (5P20GM109098), and National Institute of General Medical Sciences Grant U54 GM‐104942.
Kv1.3 and Kv1.5 Channels as Novel Targets for the Prevention of Restenosis in Human Vessels
Authors: Perez‐Garcia M Teresa , Arevalo‐Martinez Marycarmen , de la Fuente Miguel Angel , Simarro Maria , Alonso Esperanza , Lopez‐Lopez José Ramón , Cidad Pilar 
Published: November 2023

The contractile state of vascular smooth muscle cells (VSMCs) in the vessel media determines vascular tone. However, these cells can undergo a phenotypic modulation (PM) towards a synthetic phenotype in response to injury. Upon PM, VSMCs lose their contractile proteins and adquire proliferative, migratory and secretory skills. PM is central to the process of intimal hyperplasia that leads to restenosis of the vessel following vascular injury. We have previously described changes in the expression of Kv1.3 and Kv1.5 channels associated with PM. Proliferating VSMCs show an increased functional expression of Kv1.3 channels with a concomitant loss of Kv1.5 channels. Moreover, Kv1.3 blockers inhibit proliferation and migration in animal models of vascular injury and in VSMCs in primary cultures. In this work, we explored if this Kv1.5 to Kv1.3 swicth is also present in human vessels undergoing PM, and we investigate the underliying mechanisms linking Kv channel switch to the development of intimal hyperplasia.We explored PM in human mammary artery (hMA) samples using organ culture of arterial rings and primary culture of VSMCs from arterial explants. We tested the effect of Kv1.3 and Kv1.5 manipulations by overexpresion of Kv1.5 with viral vectors and blockade of Kv1.3 channels with the selective inhibitor PAP‐1. The degree of intimal hyperplasia in the organ culture was explored with hystomorphometric analyses. Changes in proliferation, migration and secretion were also studied in both preparations.Fetal Bovine Serum (FBS) treatment induced intimal hyperplasia leading to stenosis in organ culture, which was prevented with PAP‐1. PAP‐1 decreased both FBS‐induced proliferation and VSMCs secretion, as the increase in the number of elastic laminae induced by FBS was fully reverted in the presence of PAP‐1. Kv1.3 blockers also inhibited proliferation of hMA VSMCs in culture. In addition, overexpression of Kv1.5 channels had an antiproliferative effect that can be explained by Kv1.5 occlusion of Kv1.3 pro‐proliferative effects.We show that organ culture of human vessels represents a valuable approach for the study of the complex, multifactorial process of vascular restenosis. Many of these processes can be targeted by the use of PAP‐1, so that therapies using Kv1.3 blockers could represent a significant improvement for the prevention of intimal hyperplasia.Support or Funding InformationSupported by grants ISCIII‐FEDER RD12/0042/0006, (Heracles Program), BFU2013‐45867‐R (MINECO) and by UVA‐Santander fellowship to MAM.
Gender Bias in G Protein‐Coupled Receptor Expression in Human Pulmonary Artery Smooth Muscle Cells
Authors: Iyinikkel Jean , Saunders Fiona , Walker Claire , Murray Fiona 
Published: November 2023

Pulmonary arterial hypertension (PAH) is characterized by increased pulmonary vascular resistance and proliferation of pulmonary artery smooth muscle cells (PASMC). Female bias of PAH (ratio of 4.1:1, female vs. male) is well known, however not fully understood. G protein‐coupled receptors (GPCRs) are the largest family of receptors and important regulators of PASMC function. Profiling GPCRs in female‐ vs. male‐PASMC could advance the understanding of the disease and highlight the role of novel GPCRs in the female bias of PAH. We used an “unbiased” approach (a TaqMan® GPCR array) to profile GPCR expression in primary human male‐ and female‐PASMC. We found that male‐PASMCs express 190 GPCRs (42 orphan and 148 non‐orphan, n=3), while female‐PASMC express 174 GPCRs (44 orphan and 130 non‐orphan, n=3). GPCR expression correlates with function: for example Gαs‐coupled GPCRs with formation of cAMP and inhibition of cell proliferation, thus documenting that the GPCR array identifies physiologically relevant GPCRs. We found female‐PASMC have an increased expression of 25 GPCRs (>2‐fold) and a decreased expression (>2‐fold) of 19 GPCRs, compared to male‐PASMCs. Chemokine receptor 1 (CCR1) and the purinergic receptor, P2Y12, both of which are Gαi­‐coupled, were highly upregulated in female‐PASMC (21‐ and 62‐fold, respectively). GPR75, an orphan GPCR whose expression was also increased in female‐PASMC (confirmed by real‐time PCR), was up‐regulated by 17β‐estradiol in a dose‐dependent manner (0.1 nM‐1 μM, 24 h). Taken together these data provide evidence that GPCR expression in PASMC is gender specific and such differences may contribute to the physiology of normal PASMC and the female bias of PAH.Support or Funding InformationATS foundation and Pulmonary Hypertension Association and University of Aberdeen
Hypoxia‐induced proliferation and migration in pulmonary arterial smooth muscle cells requires β‐catenin.
Authors: Yun Xin , Jiang Haiyang , Shimoda Larissa A 
Published: November 2023

Exposure to hypoxia causes proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) in the distal vasculature, leading to structural remodeling of these arteries and contributing to the pathogenesis of pulmonary hypertension. In past studies, we showed that expression of the water channel, aquaporin 1 (AQP1), was upregulated by both in vivo and in vitro exposure to hypoxia, and that this channel mediated PASMC proliferation and migration. The effects of AQP1 were independent of channel water transport, but were associated with increased expression of β‐catenin, a dual function protein that is involved in the Wnt signaling pathway. β‐catenin activates transcription of genes involved in cell growth and survival and regulates the coordination of cell‐cell adhesion. Thus, we explored whether β‐catenin was required for hypoxia‐induced PASMC migration and proliferation. Primary cultures of rat PASMCs were harvested from distal pulmonary arteries and exposed to in vitro hypoxia (4% O2 ‐ 5% CO2) or control conditions. Proliferation and migration were measured using BrdU incorporation and transwell filters, respectively. Exposure to hypoxia for 24–72 h increased PASMC migration and proliferation and β‐catenin protein, but not mRNA, levels. Silencing β‐catenin in PASMCs had no significant effect on cell function under control conditions, but prevented hypoxia‐induced proliferation and migration. Based on our findings, we conclude that increases in β‐catenin protein stability/synthesis, perhaps mediated by AQP1, are crucial for hypoxia‐induced PASMC proliferation and migration.Support or Funding InformationThis work was funded by HL‐07963 and AHA0755479.
Hypoxia Alters Ubiquitination of Proteins Associated with Cellular Proliferation in Mouse Lung
Authors: Zhao Jingru , Wade Brandy , Ma Jing , Hart C. Michael , Sutliff Roy L 
Published: November 2023

Pulmonary hypertension (PH) is a chronic condition characterized by increased pressure in the pulmonary vasculature. PH is caused by constriction and/or thickening of arterial walls due in part to cellular proliferation. The underlying mechanisms for PH pathogenesis are not well understood. The ubiquitin proteasome system (UPS) regulates cell growth and protein expression by marking proteins for degradation by attaching one or more ubiquitin moieties to lysines within target proteins. We hypothesized that hypoxia, a common stimulus for PH, would alter the UPS and pathways that regulate pulmonary vascular cell proliferation in PH. The Cell Signaling PTMScan® Ubiquitin Remnant Motif (K‐ɛ‐GG) Kit was used to determine the effect of hypoxia on protein ubiquitination by mass spectrometry (MS). Mice were exposed to normoxia (21% O2) or hypoxia (10% O2) for 3 weeks, and PH was confirmed by measuring right ventricular systolic pressure. Lungs were excised for MS analysis. The MS characterization of ubiquitinated proteins revealed that hypoxia significantly changes (fold change threshold of ≥±1.5) ubiquitination of 198 peptides. Hypoxia significantly increased ubiquitination of 14‐3‐3 eta at a single lysine, and RhoA and AnnexinA2 had increased ubiquitination at two lysines. Hypoxia also significantly decreased ubiquitination of Profilin‐1 at two lysines. These proteins have been studied in other cell types and diseases and are known to regulate cellular proliferation. 14‐3‐3 eta is a chaperone protein that interacts with proteins to promote cellular migration and proliferation in lung cancer cells. RhoA regulates cell division and is downregulated in Group 2 PH. AnnexinA2 expression is increased in hypoxia‐exposed osteoblastic cells, andProfilin‐1 expression is decreased in invasive and metastic breast cancer. We conclude that hypoxia‐induced changes in ubiquitination of 14‐3‐3 eta, RhoA, Profilin‐1, and Annexin A2 may play an important role in PH pathogenesis and regulation of pulmonary vascular cell proliferation in PH.Support or Funding InformationNIH RO1HL102167 (CMH and RLS), Merit Review from VA Office of Research and Development (1I01BX001910 to CMH), and AHA postdoctoral fellowship (16POST30930007 to BEW), SIRE (to JZ)
The role of microtubule–associated protein EB3 in Pathogenesis of Pulmonary Hypertension.
Authors: Atwal Gurdeep , Sun Mitchell , Yang Xiaoyan , Sun Ying , Chen Jiwang , Komarova Yulia 
Published: November 2023

Hypoxia is one of the most powerful triggers of Pulmonary Hypertension (PH) that acts directly on endothelial cells (ECs). Translocation of hypoxia inducible factors (HIFs) from the cytoplasm into the nucleus of ECs plays a critical role in the pathogenesis of PH by inducing transcription of genes containing hypoxia response elements (HREs) such as endothelin1 (ET1), which in turn, promotes vasoconstriction of smooth muscle cells. The role of microtubule (MT) cytoskeleton in the pathogenesis of pulmonary hypertension is not yet understood. Some studies suggest that MT dynamics play a critical role in nuclear translocation of HIFs, and therefore HIF function, but the results of these studies are controversial. Here, we investigated the role of end binding protein (EB3) in endothelial cells in the pathogenesis of PH through stabilization and/or nuclear translocation of HIFs in pulmonary arterial endothelial cells. In order to understand the specific role of EB3 in ECs, we developed tamoxifen‐inducible cell specific knock out (i‐KO) mouse models, allowing deletion of Mapre3 (EB3) gene in either endothelial or smooth muscle cells (SMCs). Exposure of EB3fl/fl (control) or iSMS‐KO mice to 10% hypoxia for four weeks resulted in a significant increase in right systolic ventricular pressure (RSVP) from 22±0.8 mmHg (normoxia) to ~35±0.8 and ~33±1 mmHg, respectively. This increase in RSVP was significantly mitigated in EB3 iEC‐KO mice as compared to the other two groups (p<0.0008). The protective effects of EB3 KO in ECs were even more evident when wall thickness of small arteries was compared between groups (p<0.0001). EB3 KO mice also showed decreased HIF2α immunofluorescence in nuclei of endothelial cells in lungs indicating the critical role of EB3 in HIF2α nuclear transport. Furthermore, our experiments in human pulmonary aorta endothelial cells expressing HIF2α mutants, which are not sensitive to proteolytic degradation in normoxia, conformed the role of EB3 in HIF2α trafficking. We conclude that MT cytoskeleton and EB3 are potential targets to limit HIF function in pathogenesis of PH.Support or Funding InformationR01 HL103922