bluuebunny/crossref_metadata_2025_split huggingface dataset:
Involvement of TRPM4 in Stretch‐Induced Modulation of Lymphatic Pumping
Authors: von der Weid Pierre‐Yves , Cho Eun‐Buall , Kratofil Rachel , Lee Stewart 
Published: June 2021

Rhythmic and phasic contractions of collecting lymphatic vessels are critically involved in tissue fluid homeostasis and immune response. This contractile activity, or lymphatic pumping, is exquisitely regulated by luminal fluid pressure. However, the molecular contributors of stretch‐induced lymphatic pumping are not known. Some members of the TRP channel family, in particular TRPM4,have been shown to respond to stretch or pressure and to contribute to myogenic vasoconstriction and vascular response to stretch.Using pressure myography, electrophysiology, pharmacology and quantitative real‐time PCR, we tested the hypothesis that TRPM4 is also a major contributor of the lymphatic vessel response to stretch.We demonstrated the expression of TRPM4 mRNA in rat mesenteric lymphatic vessels and showed that the contraction frequency of the vessels pressurized at 3 mmHg was inhibited in a concentration‐dependent manner by the TRPM4 blocker 9‐phenanthrol (IC50=1.2 µM). The blocker (10 µM) also inhibited the pressure‐induce increase in contraction frequency in an endothelium‐independent manner and caused a hyperpolarization of the lymphatic muscle. These findings suggest an important role for TRPM4 in the ability of lymphatic vessels to adapt their contractile activity to changes in intraluminal lymph pressure.
Methylene Blue Mitigates Ethanol Exacerbation of Ischemia‐Reperfusion Injury of Cardiomyocytes and Brain Cells
Authors: Ryou Myoung‐gwi , Winters Ali , Robert Mallet , Yang Shaohua 
Published: June 2021

Ethanol (EtOH) intoxication is a major risk factor for myocardial and brain ischemia. Methylene blue (MB) could mitigate EtOH exacerbation of ischemia‐reperfusion injury.MethodsHT22 cells, primary microvascular endothelial cells (MVEC), astrocytes and cardiomyocytes (CM) were treated with 100 mM EtOH ± 100 nM MB for 5 days prior to 4 h oxygen and glucose deprivation (OGD, 0.1% O2) and 24 h reoxygenation. Cell viability was assayed with Calcein AM. FITC‐Dextran leakage and transendothelial electrical resistance evaluated integrity of an in vitro blood brain barrier (BBB) model composed of MVEC and astrocytes. Generation of reactive oxygen species (ROS) was monitored with mitosox stain and DCF‐DA assay. ATP concentration was measured by spectrophotometry. Aquaporin 4 and hypoxia inducible factor‐1α (HIF‐1α) contents were assessed by immunoblot.ResultsEtOH was severely detrimental to both cell viability and BBB integrity vs. OGD control. EtOH significantly increased ROS in a manner blunted by MB. ATP depletion during OGD in HT22 cells and CM was appreciably worsened by EtOH, but MB cotreatment restored ATP content to that of EtOH‐free OGD. EtOH decreased HIF‐1α and increased aquaporin 4.ConclusionEtOH increased the vulnerability of all four cell types to OGD‐reoxygenation insult. MB mitigated this EtOH toxicity by minimizing ROS generation, increasing ATP availability, and maintaining HIF‐1α, a critical factor for cellular adaptation to the ischemic environment.
Role of endoplasmic reticulum stress and thrombosis in type 2 diabetes‐induced vascular dysfunction
Authors: Ait Aissa Karima , Kassan Modar , Trebak Mohamed , Belmadani Souad , Matrougui Khalid 
Published: June 2021

Endoplasmic reticulum (ER) stress and thrombosis are important mechanisms that underlie many of the serious consequences of type 2 diabetes. However, the interaction between the ER stress and thrombosis in diabetes‐induced endothelial dysfunction remains unknown. Ten‐weeks‐old type 2 diabetic mice (db‐/db‐) and their controls (db‐/db+) treated with or without the ER stress inhibitor (tauroursodeoxycholic acid, TUDCA, 150 mg/kg per day, for 2 weeks) were used to determine the in vivo contribution of ER stress in vascular dysfunction. Blood pressure was similar in all groups of mice. Blood glucose and body weight were reduced in (db‐/db‐) mice treated with TUDCA. The impaired endothelium‐dependent relaxation (EDR), in response to acetylcholine, in (db‐/db‐) mice was restored after treatment with TUDCA in both aorta and mesenteric resistance arteries (MRA). Additionally, ER stress markers (CHOP and ATF4), NADPH oxidase activity and the mRNA levels of Nox2 and Nox4, which were increased in MRA and aorta from (db‐/db‐) mice, were significantly reduced after TUDCA treatment. Moreover, the vasoconstriction to thrombin and the mRNA levels of thrombin receptor (PAR1) were increased only in MRA from (db‐/db‐) mice and reduced after treatment with TUDCA.This study unveiled novel role of ER stress in the regulation of thrombin signaling and therefore vascular dysfunction in type 2 diabetes.
Melatonin Inhibit the Angiogenesis of Gastric Cancer by Nuclear Receptor
Authors: Zhou Ruixiang , Wang Rixiong , Song Jun , Zhang Hui , Luo Jianhua , Liu Hui 
Published: June 2021

This study aims to explore the relationship between melatonin (MLT) and gastric cancer angiogenesis by nuclear receptor with ELISA, real‐time quantitative RT‐PCR, IHC, Western blot, and siRNA methods.1. The anti‐angiogenic function of MLT on the human gastric cancer cells and the impact on MLT nuclear receptor in vitro. MLT was time‐dependent in inhibiting proliferation of human gastric cancer cell lines MGC‐803, SGC‐7901, and AGS; Chemical hypoxia simulated by 100uM CoCl2 increased proliferation of SGC‐7901 cells; MLT decreased the expression of VEGF mRNA and protein of SGC‐7901; MLT decreased the expression of nuclear receptor RZR/RORγ mRNA and protein of SGC‐7901.2. The anti‐angiogenic mechanism of MLT on human gastric cancer cells SGC‐7901 in vitro and the targets of MLT nuclear receptor RZR/RORγ. The RZR/RORγ silent by siRNA antagonized significantly MLT's inhibition on gastric cancer cell proliferation.After the 24h intervention by 3mM MLT, the mRNA and protein expression of RZR / RORγ,SENP1,HIF‐1α and VEGF was significantly decreased; but the expression of SENP1, HIF‐1α, VEGF mRNA and protein was increased.3. The anti‐angiogenic mechanism of MLT on the gastric cancer in vivo with nude mice.We established the tumor‐bearing nude mice model of gastric cancer SGC‐7901 with intervention by different concentrations of MLT. MLT reduced the tumor volume and weight of the gastric cancer; MLT inhibited tumor proliferation and angiogenesis by decreasing the expression of RZR / RORγ ,SENP1 ,HIF‐1α, VEGF and MVD.The present study confirmed that MLT is able to inhibit gastric cancer angiogenesis both in vivo and in vitro. It's major mechanism is reducing the expression of nuclear receptor RZR/RORγ and decreasing the expression of SENP1,HIF‐ 1α,VEGF.
Placental Growth Factor (PlGF) and Vascular Endothelial Growth Factor (VEGF) Receptors Expression in the Placenta – Correlation with Maternal Blood Hemoglobin Level and Pregnancy Outcome
Authors: Szukiewicz Dariusz , Stangret Aleksandra , Watroba Mateusz 
Published: June 2021

Insufficient tissue oxygenation may be caused by lowered hemoglobin (Hb) concentration in blood. Therefore, anemia during pregnancy may stimulate angiogenesis in the placental tissues. Here we examined correlations between mean placental vascular density, local expressions of PlGF and VEGF receptors (flk‐1, flt‐1), and the birth weight and maternal Hb level. After normal full‐term deliveries ending pregnancies with maternal anemia (Group I; N = 23) and normal Hb level (Group II; N = 23), tissue samples were collected from the maternal side of each placenta (3 samples per each placenta). The mean expressions of immunostained flk‐1 and flt‐1 were analyzed by quantitative morphometry in the paraffin sections. The mean vascular/extravascular tissular index (V/EVTI) was examined as a morphometric parameter which reflects intensity of vascularization by assessing a total vascular area within standardized visual fields. Nonparametric Mann‐Whitney U‐test and Spearman's correlation were used for comparative analysis of the studied groups. Among gravidas with lowered Hb level, the local expression of flt‐1 was significantly increased (p < 0.05) and positively correlated with the mean birth weight. The same association was denoted for V/EVTI. Thus, compensatory changes in the vascular system of the human placenta evoked by low maternal Hb concentration include increased angiogenesis to maintain normal growth of the fetus. Up‐regulation of flt‐1 may play a key role.WUM grant: 2M2‐W1‐14
Baroreflex impairment in a rat model of Parkinson's disease
Authors: Cavalleri Marina , Barna Barbara , Moreira Thiago , Takakura Ana 
Published: June 2021

Parkinson's disease (PD) exhibit motor and non‐motor symptoms. Among non‐motor symptoms, the cardiovascular autonomic dysfunction is frequently. The aim of this work was to evaluate whether in a model of PD induced by injection of the toxin 6‐hydroxy‐dopamine (6‐OHDA) into the striatum was able to produce changes in the baroreflex as well as in neurons in brainstem areas enrolled in cardiovascular control. Wistar rats (300‐320 g, n = 5) received bilateral injections of 6‐OHDA or vehicle into the striatum. After 60 days,baroreflex was assessed by determining the heart rate (HR) responses to ramped infusions of phenylephrine (PE) and sodium nitroprusside (SNP)induced mean arterial pressure (MAP) changes. After, the rats were anesthetized for brain perfusion and processed for tyrosine hydroxylase (TH) and the transcription factor Phox2b immunoreactive. PE produced a similar increased in MAP in 6‐OHDA and control groups. However, the bradycardia was blunted in 6‐OHDA (D = ‐7.5±0.5 to ‐62±6 vs. vehicle: ‐29.5±1.2 to ‐148.5±9.8 bpm). In 6‐OHDA, the SNP produced a smaller decrease in MAP (Δ = ‐18±2.6 to ‐24.5±0.8 vs. vehicle: ‐60.3±2 to ‐96.5±1.9 mmHg) and a smaller increase in HR (Δ = 34.75±3.4 to 59.5±7.8 vs. vehicle: 61±7.8 to 101.3±6 bpm). Ocurred a reduction in TH neurons in the substantia nigra (85%), in the C1 regions (70%) and in the Phox2b+ neurons in the NTS (48%). We observed an impairment of baroreflex sensitivity in the 6‐OHDA model of PD, which could be correlated with a reduction in the neuronal cytoarchitecture of brainstem regions involved in cardiovascular controlFinancial support: FAPESP, CNPq, CAPES/PROEX
Nitrous Oxide Inhalation Attenuates an Increase in Muscle Sympathetic Nerve Activity during Cold Stress
Authors: Isono Kazushige , Okada Yoshiyuki , Mitsui Tatsuhisa , Masuda Yuji , Ogasawara Tadashi 
Published: June 2021

Nitrous oxide inhalation was reported to suppress pressor response to pain stimulation, while an increase in heart rate (HR) remained unchanged. We evaluated whether nitrous oxide inhalation attenuates an increase in muscle sympathetic nerve activity (MSNA) during cold stress. MSNA (microneurography), arm cuff and beat‐to‐beat blood pressure (BP), and HR were measured during 5‐min baseline, 2‐min cold pressor test (CPT) in 12 young men [30±1 (SE) yrs] after they receive room air, 100% oxygen, 30%, and 40% nitrous oxide for 20 min through a mask, respectively. BP increased during CPT in all conditions, while the increase of systolic BP tended to be smaller during inhalation of 40% nitrous oxide than room air (26±5 vs 15±4 mmHg, P=0.081). HR also increased during CPT without any difference the increase between the conditions. MSNA burst frequency increased during CPT, while the increase was smaller during the inhalation of 40% nitrous oxide than room air (21±4 vs 12±3 burst/min, P=0.045). Change (Δ) in systolic BP during CPT was correlated with ΔMSNA burst frequency (r=0.40), but not ΔHR (r=0.06) in all conditions. These results suggest that suppressed pressor response to pain stimuli by inhalation of nitrous oxide is attributable to the attenuation of increase in muscle sympathetic nerve activity.Supported by JSPSKAKENHI70566661.
Aquaporin‐1 retards renal cyst development in polycystic kidney disease by inhibition of Wnt signaling
Authors: WANG Weiling , LI Fei , ZHOU Hong , Verkman Alan , YANG Baoxue 
Published: June 2021

ObjectiveAutosomal dominant polycystic kidney disease (ADPKD) is one of the most common human monogenic diseases. Aquaporin‐1 (AQP1) is expressed mainly at epithelial cell plasma membranes in renal proximal tubules, as well as in epithelial cells lining the cysts in ADPKD kidneys, postulating that AQP1 may be involved in cytogenesis. Here we investigated the role of AQP1 on ADPKD development.Methods: We used Aqp1/Pkd1 double knockout mice, embryonic kidney organ cultures, and matrix‐grown MDCK cells in this study to investigate the role of AQP1 in ADPKD. Expression of Wnt signaling‐related proteins were detected in MDCK and AQP1‐MDCK cells. Co‐immunoprecipitation was used to determine the interaction between AQP1 and β‐catenin,GSK3β,LRP6 and Axin1.Results: Kidney size and cyst number were significantly greater in AQP1‐null PKD mice than in AQP1‐expressing PKD mice. In embryonic kidney cultures, AQP1 deletion increased cyst development. In matrix‐grown MDCK cells, AQP1 inhibited cystogenesis and promoted tubulogenesis. AQP1 decreased β‐catenin and cyclinD1 expression, suggesting down‐regulation of Wnt signaling. Co‐immunoprecipitation showed AQP1 interaction with β‐catenin, GSK3β, LRP6 and Axin1. Biochemical analysis revealed decreased p‐β‐catenin and increased β‐catenin expression in Aqp1/Pkd1 double knockout mice, suggesting enhanced Wnt signaling.ConclusionsThese results suggest that AQP1 may be involved in maintaining the stability of the “destruction complex” to promote β‐catenin phosphorylation and suppress the Wnt signaling. This study reveals a previously unrecognized role of AQP1 in ADPKD and hence may provide new therapeutic targets.
Transcriptome Profiling of Human Prostate Cancer Cell Line LNCaP Treated with IndividualSoy Isoflavones or Their Combination
Authors: Jang Chan Ho , Schmidt Carl , Wu Changqing 
Published: June 2021

Prostate cancer (PCa) is the second leading cause of cancer death in American men. Increasing epidemiologic evidence and laboratory studies suggest soy isoflavones might reduce PCa risk. However, attention has been drawn to the safety of using high levels of soy isoflavones in human, which is especially the concern about taking soy isoflavone dietary supplements. Our previous work showed that daidzein and genistein had a synergistic effect on inhibiting proliferation and inducing apoptosis of PCa cells, which suggests a combination of isoflavones at lower levels may be more effective for PCa prevention. Based on this work, RNA sequencing (RNA‐seq) was performed in this study to search underlying mechanisms in human prostate cancer cell line LNCaP treated with individual isoflavones (genistein or daidzein, 50 or 200 μM) or their combination (50 μM). Transcriptome analysis showed that several genes were differently expressed in LNCap cells among treatments with individual isoflavones or their combination.Genes such as transcription factor CP2‐like 1, activating transcription factor 3, PRKC apoptosis WT1 regulator, DNA‐damage‐inducible transcript 3 and interferon gamma‐inducible protein 16 are highly expressedin the cells treated with genistein orthecombination of daidzein and genistein, while not highly expressed in the cells treated only with daidzein. To conclude, these findings have indicated that RNA‐seq is valuable to simultaneously search multiple molecular targets involved in the cancer preventive effect of soy isoflavones and a reliable means to quantify absolute transcript levels. The findings might also help explain the synergistic effect of daidzein and genistein.The project is funded by University of Delaware Research Foundation.
Reduced Renal K+ Excretion With Compensatory Hyperaldosteronism In KCa1.1 Channel β2‐subunit KO Mice
Authors: Larsen Casper , Sorensen Mads , Praetorius Helle , Leipziger Jens 
Published: June 2021

The kidneys are the primary organs responsible for excreting K+, ensuring whole body K+ homeostasis by precisely matching K+ excretion to dietary K+ intake. K+ is secreted into the urine in the collecting ducts, and two distinct mechanisms for K+ secretion exist; a constitutive mechanism mediated by ROMK (Kir1.1) in principal cells and a flow‐induced mechanism mediated by BK channels (KCa1.1) in intercalated cells. Both mechanisms are up‐regulated by aldosterone. Here we studied renal K+ excretion in KO mice for the β2‐subunit of the BK channel.The β2 KO mice have increased plasma aldosterone, low renin expression and normal plasma [K+]. The low renin in β2 KO mice indicates that hyperaldosteronism was triggered by a K+ handling deficiency, rather than hypotension and activation of the renin‐angiotensin‐aldosterone system. We hypothesize that β2 KO mice have decreased BK channel‐mediated renal K+ secretion, which is compensated by hyperaldosteronism and up‐regulation of ROMK‐mediated K+ secretion, allowing β2 KO mice to maintain normal plasma [K+]. In fact, when treated with eplerenone (mineralocorticoid receptor antagonist) for 4 days, β2 KO mice develop slight hyperkalemia (4.15 mM ± 0.13 in WT vs. 4.60 mM ± 0.10 in KO, P = 0.013).Urinary K+ excretion following oral K+ load (20% of normal daily intake) was not different between WT and KO mice under control conditions. However, when treated with eplerenone, β2 KO mice had a significantly lower urinary K+ excretion rate (P=0.044) and significantly higher plasma [K+] 3 hours after oral K+ load (10.0 mM ± 0.4 in WT vs. 11.4 mM ± 0.5 in KO, P = 0.044). Our data support that hyperaldosteronism in β2 KO mice is part of a chronic compensation to a decreased BK channel‐mediated renal K+ secretion.Supported by the Lundbeck foundation.
Placental Ischemia‐Induced TH17 Cells Mediate the Pathophysiology Associated with Preeclampsia
Authors: Cornelius Denise , Wallace Kedra , Amaral Lorena , Scott Jeremy , Campbell Nathan , Thomas Alexia , Moseley Janae , LaMarca Babbette 
Published: June 2021

Preeclampsia (PE) is associated with hypertension, inflammation, intrauterine growth restriction (IUGR), increased CD4+ THelper 17 cells (TH17s) and agonistic auto‐antibodies to the AT1 receptor (AT1‐AA). The objective of this study was to determine a role for TH17s in mediating pathophysiology associated with PE using the reduced uterine perfusion pressure (RUPP) rat model of PE. On gestation day 12 (GD12) RUPP‐induced TH17s were injected into normal pregnant (NP) rats; On GD19 blood pressure (MAP) was recorded, and blood and tissues were collected. One‐way ANOVA was used for statistical analysis. MAP increased from 99±2 in NP (n=8) to 128±4 in RUPP (n=9) and to 111±3 mmHg in NP+TH17 (n=11; p<0.05). Placental and renal ROS increased from 238 and 411 RLU, respectively, in NP to 328 and 603 RLU in RUPP and to 339 and 833 RLU in NP+TH17 (p<0.05). Importantly pup weight (gm) decreased significantly from 2.19 in NP to 1.91 in RUPP, and to 1.9 in NP+TH17 (p<0.05). Adoptive transfer of TH17s also increased inflammation: IL‐6 increased significantly from 27.5 pg/mL in NP to 62.2 pg/mL in RUPP and to 75.9 pg/mL in NP+TH17 (p<0.5); IL‐17 increased from 0.5 pg/mL in NP to 1.29 pg/mL in RUPP and increased to 5.5 pg/mL in NP+TH17 (p>0.05). AT1‐AA significantly increased from 0.1beats/min in NP to 16.82beats/min in RUPP and to 15.6beats/min in NP+TH17. TH17s induced hypertension, ROS, IUGR, inflammation, and AT1‐AA in pregnant rats, demonstrating the importance of TH17s to mediate the pathophysiology associated with PE and their potential as a new therapeutic target for treatment of PE.NIH grants RO1HD067541 and T32HL105324
N‐Acetyl‐Seryl‐Aspartyl‐Lysyl‐Proline (Ac‐SDKP) Delays the Development of Hypertension and Renal Damage in Systemic Lupus Erythematosus (SLE)
Authors: Nakagawa Pablo , Liao T‐D , Worou M , Basha H , Peterson E , Janic B , Yang X‐P , Rhaleb N‐E , Carretero Oscar 
Published: June 2021

SLE is an autoimmune disease with high prevalence of hypertension. Ac‐SDKP is naturally occurring peptide with anti‐inflammatory and immunomodulatory properties. In classical hypertension models, Ac‐SDKP prevents renal damage without affecting systolic blood pressure (SBP). We hypothesize that in autoimmune systemic diseases associated with hypertension, such as SLE, in addition to preventing renal damage, Ac‐SDKP also prevents the development of hypertension. Here we used NZBWF1 mice as SLE hypertension model. Female NZW (control) or NZBWF1 (SLE) mice were divided in 4 groups: 1) control+vehicle, 2) control+Ac‐SDKP, 3) SLE+vehicle, 4) SLE+Ac‐SDKP. Animals were treated with vehicle or Ac‐SDKP (800 µg/kg/d via minipumps) from 25 to 38 wks of age. Urine albumin and SBP were recorded weekly. Plasma anti‐dsDNA antibodies were measured at 38wks. At 28 wks of age 40 % of the animals in the SLE+vehicle group developed albuminuria (defined as urine albumin 蠅 30 µg/24hs), while in the SLE+Ac‐SDKP group the incidence of albuminuria was 10 %. At 32 wks of age the incidence of hypertension (defined as SBP 蠅 140 mmHg) in SLE+vehicle was 40 %, while none of the animals in SLE+Ac‐SDKP group developed hypertension. At 32 wks, the survival rate was 60 % in SLE+vehicle group and 90% in SLE+Ac‐SDKP 90 % group. However, at 38 wks of age the incidence of albuminuria, hypertension, mortality and plasma dsDNA antibody levels were similar between SLE+vehicle and SLE+Ac‐SDKP groups. In SLE, Ac‐SDKP delays the incidence of renal damage, hypertension and mortality without affecting the dsDNA antibody levels. Supported by NIH HL028982 and AHA 13PRE17100074
Expression of claudin‐2 in the uroepithelium increases urinary bladder activity
Authors: Montalbetti Nicolas , Kullmann F Aura , Rued Anna , Clayton Dennis , Prakasam H Sandeep , Ruiz Wily , Apodaca Gerard , Carattino Marcelo 
Published: June 2021

Bladder filling promotes a reorganization of the umbrella cell tight junctions (TJ) ring, which is accompanied by a large drop in junctional resistance. The goal of this study was to assess the impact of increased paracellular permeability on bladder function. Claudins are structural components of the TJ that define the permeability of the paracellular barrier present in epithelia. To increase the paracellular permeability of the rat uroepithelium we expressed claudin‐2 using in situ adenoviral transduction. Fluorescent microscopy showed prominent expression of claudin‐2 in the umbrella cells of the transduced bladders. The electrical resistance and potential difference of the urinary bladders mounted in Ussing chambers were significantly lower in rats transduced with claudin‐2, 0.5 ± 0.1 kΩ.cm2 and ‐1.3 ± 0.2 mV, than in rats transduced with GFP, 4.6 ± 0.3 kΩ.cm2 and ‐12 ± 2 mV. Conversely, the permeability of the uroepithelium to large molecules (MW >350), fluorescein and NHS‐biotin, was similar in rats transduced with GFP and claudin‐2. In vivo studies of bladder function using continuous cystometry revealed higher intravesical basal pressures and shorter intervoiding intervals in rats transduced with claudin‐2 than in rats transduced with GFP. In conclusion, the expression of claudin‐2 in the uroepithelium increases its permeability to ions and augments voiding frequency, consistent with the notion that paracellular ion transport modulates bladder function.Support: NIH R01 DK099196 and P30 DK079307 Pittsburgh Center for Kidney Research ‐ O'Brien Pilot.