bluuebunny/crossref_metadata_2025 huggingface dataset:
The Interplay between CCR1 and IL‐6 in Osteoclasts
Authors: Habbal Renee , Sakadjian Nishan , Gilchrist Annette
Published: November 2023

Osteoclasts are highly specialized multinucleated giant cells responsible for the process of bone resorption. The chemokine receptor CCR1 is thought to be involved in inflammatory bone destruction through its ability to enhance osteoclastogenesis. Using the murine macrophage cell line RAW264.7, which can be differentiated into osteoclasts by addition of receptor activator of nuclear factor κB ligand (RANKL), we examined several pharmacological inhibitors of CCR1 for their ability to displace 125I‐CCL3 binding and influence IL‐6 production. The role of IL‐6 in osteoclastogenesis has been debated, with some researchers suggesting IL‐6 stimulates osteoclast differentiation while others propose IL‐6 inhibits this process. We found that all of the compounds tested were able to displace 125I‐CCL3 binding. However, they differed with respect to their ability to inhibit IL‐6 production with some but not all resulting in increased IL‐6 production from RANKL‐differentiated RAW264.7 cells. Together, our data supports prior findings that the CCR1 antagonists are biased, and that IL‐6 inhibits RANKL‐induced osteoclastogenesis.
Andrographolide restores healthy airway function in toluene diisocyanate‐induced occupational mouse asthma model through upregulation of heam oxygenase‐1 and activation of Akt/Nrf‐2 dependent pathway
Authors: Sulaiman Ibrahim , Tan Khaishin , Mohtaruddin Norhafizah , Lim Jonathan Chee Woei , Stanslas Johnson
Published: November 2023

Toluene diisocyanate (TDI) is a major cause of chemical‐induced occupational asthma, which contributes about 15% of global asthma burden. Steroid resistance and compounded side effects that are associated with use of corticosteroid in management of asthma necessitate the search for alternative drugs. Andrographolide, a naturally occurring diterpene lactone was tested for control of cardinal occupational asthma features. BALB/c mice were dermally sensitized with 0.3% TDI or acetone olive oil (AOO) vehicle on day 1 and 8, followed by 0.01% TDI intranasal challenge on days 15, 18 and 21. Endpoints were evaluated via bronchoalveolar lavage fluid (BALF) cell analysis, 2′,7′‐dichlorofluorescin diacetate (DCFH‐DA) intracellular reactive oxygen species (ROS) assays, immunoblotting, immunohistochemistry and methacholine challenge tests. Decrease in BALF total and differential leukocyte counts was recorded in the andrographolide‐treated animals. The compound dose‐dependently (at 0.1mg/kg, 0.5mg/kg and 1mg/kg) reduced intracellular de‐esterification of DCFH‐DA. This suggests andrographolide's role in the neutralization of intracellular BALF ROS surge. Mechanistically, the treatment prevented TDI‐induced aberrant E‐cadherin bronchial epithelial distribution at cell‐cell contact site. Immunoblot analysis of lung samples shows significant up‐regulation of heam oxygenase‐1 (HO‐1) and nuclear factor erythroid 2‐related factor‐2 (Nrf2) which may have possibly occurred through Akt dependent pathways. Methacholine triggered airway hyperresponsiveness was markedly suppressed following andrographolide administration (i.p). Collectively, these findings indicate that andrographolide exhibit protective function against TDI induced airway epithelial barrier dysfunction and oxidative lung damage, and that Akt dependent pathways may play a role in the restoration of normal airway functionality in chemical induced occupational asthma. To the best of our knowledge, the specific ability of andrographolide to control TDI‐induced occupational asthma was not previously explored. Therefore, these data elucidate the therapeutic potential of the candidate compound in the control and management of chemical induced occupational asthma.
Anti‐inflammatory Evaluation Of LASSBio‐1823, A New p38 MAPK Inhibitor
Authors: de Carvalho Patrícia Ribeiro , de Morais Cordeiro Natália , Freitas Rosana Ribeiro , Alves Marina , Manssour Carlos Alberto Ribeiro , Fernandes Patricia Ribeiro
Published: November 2023

Protein kinases are directly involved in many human disorders, including inflammatory diseases and cancer. The p38 mitogen‐activated protein kinase (MAPK) plays a crucial role in a signal transduction cascade regulating biosynthesis and release of pro‐inflammatory cytokines. Our aim was to evaluate LASSBio‐1823 effects using in vitro and in vivo models of inflammation.

LASSBio‐1823 was evaluated in vitro p38 MAPK assay, carrageenan‐induced leukocyte migration into subcutaneous air pouch (SAP) of mice and measurement of nitric oxide (NO) and TNF‐α obtained from SAP exudates and after activation of J774.A1 macrophage cell line in vitro. J774.A1 cells were incubated with LASSBio‐1823 (1, 10 or 30 μM) and further activated with lipopolysaccharide (LPS, 1 μg/mL). After 24 h the supernatant was collected and NO and TNF‐α measured. Mice (Swiss Webster, 22–25g, n=4–6, authorization of use #DFBCICB015‐04/16 (COBEA/UFRJ/Brazil)) were orally treated with LASSBio‐1823 (10 μmol/kg) 1 h before carrageenan injection in SAP. After 24 h animals were euthanized and exudate was collected for subsequent dosages. Results are presented as mean±SD. Statistical analysis was ANOVA and Bonferroni post‐test (*p<0.05).

Results obtained in p38 enzymatic assay demonstrated LASSBio‐1823 IC50 of 21.33 μM (10.5–43.5 μM). The compound reduced NO production by LPS‐activated J774.A1 cells: 1 μM: 77.5±27 μM, 10 μM: 7.5±5.2 μM* and 30 μM: 14.6±11.2 μM* when compared to LPS: 85.5±20.1 μM. It was also observed a reduction in TNF‐α levels in LPS‐activated cells: 1 μM: 1,430.3±48.5 pg/mL, 10 μM: 865.6±53.9 pg/mL* and 30 μM: 324.0±27.2 pg/mL* when compared to LPS: 1,313.8±146.0 pg/mL. LASSBio‐1823 (10 μmol/kg, p.o.) also reduced leukocyte migration induced by carrageenan (54.3±33.0 cells × 106/mL*) when comparing with carrageenan‐injected group (128±9.9 cells × 106/mL). Levels of NO (91.7±43.0 μM*), TNF‐α (222.4±57.3 pg/mL*) and protein extravasased (69.6±21.8 pg/mL*) were reduced when compared to carrageenan‐treated groups (242.5±117.6 μM, 454.2±148.6 pg/mL and 273.0±14.7 pg/mL, to NO, TNF‐α and protein, respectively).

Our results suggest that LASSBio‐1823 reduces some parameters of an inflammatory process thus suggesting as a prototype of a new anti‐inflammatory drug.

Support or Funding Information

Financial support: CAPES, CNPq, FAPERJ and Instituto Vital Brazil (donation of animals).

Technical support: Alan Minho.
Tetrachloroethene‐mediated autoimmunity: Significance of lipid‐derived aldehydes
Authors: Wang Gangduo , Wang Jianling , Ansari G.A. Shakeel , Khan M. Firoze
Published: November 2023

Tetrachloroethene (perchloroethylene, PCE), a ubiquitous environmental contaminant, has been implicated in inducing autoimmunity/autoimmune diseases (ADs), including systemic lupus erythematosus (SLE) and scleroderma in humans. However, experimental evidence suggesting the potential of PCE in mediating autoimmunity is lacking. This study was, therefore, designed to explore PCE's potential in inducing an autoimmune response. Six‐week old female MRL+/+ mice, in groups of 6 each, were treated with PCE (0.25, 0.5 or 1.0 mg/ml) via drinking water for 12, 18 and 24 weeks and markers of lipid peroxidation (oxidative stress) and autoimmunity were determined. PCE exposure led to significant increases in malondialdehyde (MDA)‐protein adducts, and anti‐ MDA‐/4‐hydroxynonenal (HNE)‐protein adduct antibodies, which showed a time‐dependent response. The increases in anti‐MDA‐/HNE‐protein adduct antibodies were associated with significant increases in serum anti‐nuclear antibodies (ANA) and anti‐dsDNA antibodies at 18 weeks and, to a greater extent at 24 weeks, suggesting that PCE exposure exacerbated autoimmunity in our animal model. The splenocytes isolated from mice treated with PCE for 18 and 24 weeks showed greater Th17 cell proliferation and increased release of IL‐17 in culture supernatants following stimulation with MDA‐mouse serum albumin adducts, suggesting that MDA‐modified proteins may act as an immunologic trigger by activating Th17 cells and contributing to PCE‐mediated autoimmunity. Our studies provide the experimental evidence that PCE induces/exacerbates an autoimmune response and lipid‐derived reactive aldehydes (MDA and HNE) contribute to this response.

Support or Funding Information

Supported by NIH ES023106.
Changes in Bone Structure, Biomarkers and Density in Collagen‐Induced Arthritis
Authors: Samadfam Rana , Poulin Dominic , Varela Aurore , Jolette Jacquelin , Boyd Gabrielle , Cornicelli Joe , Young Alan
Published: November 2023

The purpose of this study was to evaluate the bone density using Dual X‐ray Absorptiometry (DXA), osteoclast‐derived tartrate‐resistant acid phosphatase form 5b (TRACP‐5b) in serum and osteoclast cellularity with immunostainingin a rat CIA model. Female Lewis rats were assigned to non‐disease or to collagen‐induced arthritis (CIA) groups. Under isoflurane anesthesia, animals in CIA group were injected intradermally in two sites on the back at the base of the tail (150 μL per site) with bovine type II collagen (CII) emulsified in Freund's incomplete adjuvant. This procedure was done on Day 1 and again on Day 7 (booster). Clinical score and paw inflammation were evaluated daily. DXA was used to measure bone mineral density (BMD), bone mineral content (BMC) and area using a Hologic Discovery A bone densitometer. Single scans at the lumbar spine and tibiotarsal joint were acquired once pretreatement, on Day 16 and on Day 25. Serum TRACP‐5b were measured once pretreatment, on Day 16 and Day 26. Paw tissue samples were collected for TRACP‐5b, histology and immunohistochemistry evaluations. In the CIA animals, clinical score and paw inflammation peaked on Day 16 and remained stable until Day 26. Decreases in DXA parameters (bone area, mineral content and density at the lumbar spine and bone density at the tibiotarsal joint) and serum TRACP‐5b were observed, while paw tissue TRACP‐5b, histopathological composite score and osteoclast cellularity were increased. These data clearly indicate changes in bone structure, biomarkers and density after induction of arthritis.
Anti‐cancer mechanisms of lovastatin in A549 human non‐small cell lung cancer cells
Authors: Wang Jie‐Yu
Published: November 2023

Lung cancer is a leading cause of death worldwide, and non‐small cell lung cancer (NSCLC) is the most common type of lung cancer. Although statins, 3‐hydroxy‐3‐methylglutaryl coenzyme A (HMG‐CoA) reductase inhibitors, are originally used for treatment of hypercholesterolemia, previous studies implicated that statins may have beneficial effect for the prevention or treatment of cancer. Thereby, the purpose of this study was to explore the potential effects and mechanisms of lovastatin, one of statin drugs, on chemotherapy of NSCLC. Besides, the next generation sequencing (NGS) was also applied to clarify the intracellular regulatory network of lovastatin within A549 cells. Cell viability and migration were examined by MTT and wound‐healing assays, respectively. The protein level was monitored by Western blot. Experimental data showed that lovastatin treatment can reduce AKT‐mTOR and ERK1/2‐MAPK pathway, arrest cell cycle at G0/G1 phase, as well as suppress serum‐induced A549 growth and migration. Our study suggested that lovastatin can be a potential treatment for NSCLC.
Let‐7c is the Most Effective among a Set of Bioengineered miRNA Agents in Inhibiting Hepatocellular Carcinoma Cell Proliferation
Authors: Jilek Joseph Lionel , Ho Pui Yan , Duan Zhijian , Yu Ai‐Ming
Published: November 2023

Hepatocellular carcinoma (HCC) is the most common liver cancer in the world. Few curative drugs are currently on the market, including sorafenib, although efficacy is limited and resistance and recurrence remains a significant problem. Dysregulation of microRNAs (miRNA), a family of short non‐coding RNAs, has been revealed as a significant contributor to HCC signaling, thus posing as a pharmacological target. However, current research is limited to the use of synthetic miRNA reagents that carry extensive artificial modifications. Recently our group has established a robust, fermentation based approach for the production of miRNA agents using a tRNA/pre‐miRNA scaffold. Using this approach, we have generated a small set of biologic miRNA/siRNA agents. Upon transfection, several agents were identified to significantly reduce Huh7 and Sk‐Hep‐1 cell viability. Further study on this subset of hits revealed chimeric let‐7c as the most potent and effective miRNA in inhibiting proliferation of both Huh7 and Sk‐Hep‐1 cell lines, with EC50 values of 2 and 7 nM, respectively. Subsequently, we demonstrated that biologic let‐7c significantly enhanced the sensitivities of Huh7 and Sk‐Hep‐1 cells to doxorubicin and sorafenib. These were accompanied by an 80‐fold and 20‐fold increase in mature let‐7c in Huh7 and Sk‐Hep‐1 cell lines, respectively, as well as suppression of let‐7c target gene expression (e.g., Bcl‐2/Bcl‐xL), following the transfection with biologic let‐7c. Overall, these data demonstrate that biologic let‐7c is a potent antiproliferative agent for HCC, and it also sensitizes HCC cells to sorafenib and doxorubicin.

Support or Funding Information

This study was supported by grant [R01GM133888] from NIGMS, NIH. J.L.J. was supported by a NIGMS Pharmacology Training Program [T32GM099608].
The Anti‐cancer Effects of Resveratrol on Canine Hemangiosarcoma Cells
Authors: Alderete Karen Stefania , Carlson Ariel , Grant Marianne , Sharkey Leslie , Zordoky Beshay
Published: November 2023

Hemangiosarcoma (HSA) is a highly malignant tumor of vascular endothelial origin that exhibits aggressive biological behavior. HSAs are more common in dogs than other species including cats, sheep, horses, and cattle. Generally, canine HSAs have poor prognosis. After the complete surgical excision of the primary tumor, the survival time of dogs with HSA is typically short, less than 3 months. Adjuvant treatment with chemotherapy, especially anthracycline‐based regimens such as doxorubicin, can result in modest improvement of survival rates. However, the median survival time of dogs with HSA is still short, even after chemotherapy and surgery. Therefore, there is an urgent need to improve the adjuvant chemotherapeutic regimens in order to reach better clinical outcomes in dogs with HSA. Metformin and resveratrol have been shown to have strong anti‐proliferative and/or pro‐apoptotic properties in several human cancer cell lines. However, the potential anticancer effects of metformin and resveratrol have not been determined in canine HSAs. Therefore, the objective of this study is to investigate the potential growth inhibitory effects of metformin and resveratrol on HSA cell lines when used alone or in combination with doxorubicin. FROG and DD‐1 canine HSA cell lines were treated with varying concentrations of metformin or resveratrol with and without doxorubicin. Cell viability was measured by the MTT assay, and the expression of apoptotic proteins was assessed by western blotting. In contrast to human cell line studies, metformin had a minimal inhibitory effect on the growth of HSA cells. However, resveratrol markedly inhibited the growth of HSA cells, which is in agreement with human cell line studies. In combination, resveratrol and doxorubicin inhibited the growth of both HSA cell lines more than doxorubicin alone. Mechanistically, resveratrol induced apoptosis in HSA cells, and augmented doxorubicin‐induced apoptosis in these cells. Additionally, MTT cell viability of the cardiac‐derived H9c2 cells showed that resveratrol does not augment doxorubicin‐induced cell death in cardiac cells. These findings suggest that resveratrol may have anticancer effects against canine HSA and could be a potential adjunct therapy to the standard‐of‐care anticancer drugs used in canine HSA patients. Since resveratrol has been shown to have both anti‐cancer and cardioprotective effects in rodent models, canine cancer patients can be a perfect translational platform to move the findings of these rodent studies forward to the clinic.

Support or Funding Information

Research reported in this abstract was supported by a Pre‐K Career Development Award from the National Center for Advancing Translational Sciences of the National Institutes of Health Award Number UL1TR000114. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
The β‐blocker carvedilol blocks UVB‐induced DNA damage and apoptosis of skin
Authors: Liang Sherry , Huang Kevin M. , Andresen Bradley T. , Huang Ying
Published: November 2023

β‐blockers are common pharmacological antagonists of the catecholamine activated β‐adrenergic receptors (β‐ARs). Traditionally prescribed for cardiovascular diseases such as hypertension, this class of drugs has recently been proposed with new uses in cancer treatment and prevention. Our previous studies showed that carvedilol, a nonselective GRK/β‐arrestin biased agonist, prevents skin carcinogenesis in vitro and in vivo. In the present study, we aimed at investigating the effects and mechanism of carvedilol on ultraviolet B (UVB) induced skin DNA damage and apoptosis. To assess the effects of carvedilol in vitro, the tumor promotion sensitive mouse epidermal JB6 P+ cells were treated with H2O2 or UVB to induce cell death and apoptosis. Protective effects of carvedilol was observed through cell counting, SRB cytotoxicity assay and apoptosis assay. Carvedilol effectively prevented H2O2 induced formation of reactive oxygen species in JB6 P+ cells. To explore the mechanism of action, JB6 P+ cells were treated with UVB followed by treatment with carvedilol (to avoid sunscreen effect) in various concentrations and time points. As expected, Western Blot analysis showed that the protein levels of p53 and phospho‐p53 at ser15 were induced by UVB treatment, whereas the effect of UVB on p53 was attenuated by carvedilol. We further examined the effects of topically administered carvedilol on SKH‐1 hairless mice irradiated with a single minimal effective dose (MED) of UVB (200 mJ/cm2). Our results showed that carvedilol protected UVB induced cyclobutene pyrimidine dimer (CPD) formation and apoptosis, 6 and 24 hours after UVB exposure. Consistently, the increased levels of total p53 and phospho‐p53 at Ser15 in mouse skin induced by UVB was decreased after carvedilol treatment. The change in p53 did not occur at the mRNA level, given RT‐PCR analysis failed to detect any change in the Tp53 gene. From these results, we conclude that carvedilol exhibits a protective effect against UVB induced DNA damage and apoptosis, partly related to its effect on UVB‐induced p53. These results further support the notion that the β‐blocker carvedilol may be re‐purposed for skin cancer prevention.
The Role of DNA Polymerase Nu and Zeta in the Repair of DNA Interstrand Crosslinks
Authors: Bernard Faviola , Althaus Irene , Fisher Adam , Jasty Vijay , Canman Christine
Published: November 2023

Interstrand DNA Cross‐links (ICLs) are covalent bonds between the two DNA strands. They can be induced by endogenous aldehydes produced by metabolic pathways or exogenous agents such as chemotherapeutic drugs. ICLs inhibit DNA replication and RNA transcription and are lethal if not repaired. Lesions that are not repaired may cause chromosome breakage and genome instability that could ultimately promote cancer. The Fanconi anaemia (FA) pathway is in charge of removing the ICLs by recruiting specific endonucleases to ‘unhook’ the ICL and DNA translesion synthesis polymerases (TLS) that insert and extend nucleotides opposite the unhooked ICL. Ultimately, the FA pathway directs homologous recombination repair enzymes to complete the repair process and restore the DNA to its original double stranded form. Our major goal is to identify which proteins of the FA pathway regulate TLS polymerase activity. We are currently investigating the roles DNA polymerase Nu (POLN) and DNA polymerase Zeta (REV3) play in TLS during ICL repair. Current models suggest that REV3 is required for TLS but it still unknown if POLN is also involved in the same pathway. Using RNA‐Interference, we depleted HeLa cells of POLN, REV3, or both proteins to determine if they act in the same pathway. First, we determined whether HeLa cells depleted of one or both DNA polymerases induced the similar or enhanced cytotoxicity as measured by colony forming assays following exposure to Cisplatin (CDDP) or Mitomycin‐C (MMC). Second, we examined whether double knockdown cells showed similar or elevated chromosomal aberrations following drug exposure. Third, we performed an immunofluorescence in which we stain for 53BP1 and p‐ATM, markers for DNA double strands breaks. Our preliminary data shows evidence of failure to complete ICL repair when REV3 or POLN are depleted from the cells. Together, the data suggests POLN and REV3 are involved in ICL repair.
Effect of Prenatal Methamphetamine Exposure on Blood Pressure and Vascular Function in Adult Male and Female Offspring
Authors: Harrison Allison M. , Seeley Sarah L. , Rorabaugh Boyd R. , Chrissobolis Sophocles
Published: November 2023

Hypertension and endothelial dysfunction are commonly associated with cardiovascular disease. Methamphetamine (meth) use is a widespread societal problem, with ~4.7% of the US population having tried meth at least once. Meth use during pregnancy can have deleterious developmental and behavioral consequences for offspring, including delayed development of gross motor skills and decreased memory. Although prenatal meth exposure reportedly causes increased cardiac ischemia in females, there exists very little additional data on the cardiovascular consequences of prenatal meth exposure in adult offspring. The aims of this study were to therefore test whether prenatal meth exposure increases blood pressure and impairs endothelial function in adult offspring, and whether this is influenced by gender. Pregnant female Sprague Dawley rats (8 wk old) were treated with either saline or methamphetamine (5 mg/kg/d sc, once daily) from gestational day 1 until pups were born. Male and female pups were weaned at 3 weeks of age and blood pressure (using tail cuff plethysmography) and vascular function (using myography) assessed after rats were 8 weeks old. Systolic blood pressure (SBP) was increased in male offspring prenatally exposed to meth (SBP = 151±2 mmHg, n=11) vs saline (141±3 mmHg, n=11, P<0.05 vs meth). SBP was similar in female offspring prenatally exposed to meth (SBP = 128±5 mmHg, n=7) and saline (SBP = 125±4 mmHg, n=8, P>0.05 vs meth). In males, endothelium‐dependent relaxation (assessed via relaxation responses to acetylcholine [ACh]) in second‐order mesenteric resistance arteries was impaired by prenatal meth exposure (pEC50: meth‐exposed = −7.5±0.06, n=5; saline‐exposed = −8.0±0.1, n=6; P<0.05). Prenatal meth exposure had no effect on responses to ACh in females (pEC50: meth‐exposed = −7.8±0.05, n=5; saline‐exposed = −8.1±0.08, n=5; P>0.05). Vasorelaxation responses to the endothelium‐independent agonist DEA NONOate, and phenylephrine‐induced vasoconstriction were not influenced by prenatal meth exposure in either gender. These data suggest that prenatal exposure to methamphetamine increases blood pressure and causes endothelial dysfunction in adult male offspring, whereas female offspring prenatally exposed to methamphetamine are protected against an increase in blood pressure and endothelial dysfunction. Prenatal methamphetamine exposure may therefore increase risk of cardiovascular disease in males.

Support or Funding Information

Raabe College of Pharmacy, Ohio Northern University
Gestational Exposure to Engineered Nanomaterials Effects Microvascular Health of Young Progeny
Authors: Stapleton Phoebe A
Published: November 2023

Engineered nanomaterials (ENM) are specifically manufactured at less than 100 nm in one dimension to take advantage of the unique physiochemical properties. While ENM are pervasive in domestic use, many are also being explored in biomedical applications as a platform for therapeutic or diagnostic (theranostic) applications. The toxicological implications of their use are tested primarily in healthy young adult male populations. However, these biomedical applications as contrast agents or directed drug delivery platforms may also be medically relevant during gestation. Unfortunately the outcomes during pregnancy and for developing young are unknown. In early studies, gestational ENM exposure led to impairments in uterine microvascular reactivity indicative of the development of a hostile gestational environment. The Barker Hypothesis theorizes that development is such an environment will influence cardiovascular health later in life.

Therefore, studies were initiated to assess the microvascular health of fetal and postnatal young at differing developmental time points, exposed to nano‐sized titanium dioxide through maternal inhalation during gestation. Microvascular reactivity was assessed via a pressurized isolated microvessel preparation where the arterioles of the tail (fetal [gestational day 20], neonate [6 ± 0.48d]) or heart (weanling [25.7 ± 0.9d], juvenile [7.64 ± 0.48w], adult [10.36 ± 0.34w]) progeny were dissected, excised, and evaluated in response to chemical stimuli to calculate endothelium‐dependent [acetylcholine (10−9–10−4 M)], ‐independent [spermine‐NONOate (10−9–10−4 M)], and vascular smooth muscle [phenylephrine (10−9–10−4 M)] function. Significant temporal impairments in endothelium‐dependent reactivity were identified in fetal (−37.2% ± 7.3 vs. control, respectively), juvenile (−29.1% ± 11.3) and adult (−59.3% ± 17.1) exposed progeny. ­

Following experiments performed with antioxidant treatment ([TEMPOL, 1 × 10−4 M] and catalase [50 units/mL]) in the isolated microvessel bath of the young adult cohort, led to an 86% ± 11.2 improvement in arteriolar reactivity and returned function to near control levels. These results indicate an increase in reactive oxidative stress (ROS) may be interfering in arteriolar function. Future studies may explore the origination of the ROS in the exposed progeny. Understanding the mechanisms associated with F1 effects may allow for pregnant women to safely reap the benefits of nanotechnology‐enabled products.

Support or Funding Information

NIH‐K99‐ES024783; P30‐ES005022
Antenatal Betamethasone Fetal Programming of Cardiometabolic Disease Is Reversed by Removal of Visceral Adipose Tissue Early in Life
Authors: Shaltout Hossam A , Massmann Angela , Figueroa Jorge
Published: November 2023

The use of antenatal betamethasone (BM) is a widely accepted therapy to improve survival in preterm infants. We have developed an experimental model of programming whereby pregnant ewes are administered BM (2 doses of 0.17 mg/kg) or vehicle at the 80th day of gestation and offspring deliver spontaneously at term. Our earlier studies show that BM‐exposed (BMX) offspring had a markedly impaired (~50% reduction) baroreflex sensitivity for control of heart rate (BRS) by 6 weeks of age followed by persistent suppression of the reflex and elevation in mean blood pressure (MAP) and development of glucose intolerance by 6 months of age compared to control group. In this study, we investigated the hypothesis that fetal programming of adipose tissue contributes to the elevation in MAP and reduction in BRS in BMX sheep. In a group of beta exposed sheep, Under general anesthesia, intra‐abdominal visceral fat (omental and perirenal) was surgically removed at 2–3 months of age, (BMX‐FR, n=7) and animals allowed to recover. At one year of age, heart rate and MAP were recorded continuously via femoral arterial catheters and BRS was tested using multiple doses of phenylephrine administered via femoral venous catheter to elevate MAP eliciting the reflex bradycardia. Data are presented as Mean ± SEM, and were analyzed by one way anova. While fat removal did not affect total body weight it lowered MAP by 20 mm Hg (from 95 ± 1 in BMX to 75 ± 4 in BMX‐FR, p < 0.05 to a level that is not different from the control group 78 ± 2 mm Hg). Also BRS improved by ~100 % as measured by ms/mm Hg (from 6 ± 1 in BMX to 13.8 ± 2.5 in BMX‐FR, p < 0.05 to a level that is not different from the control group 17 ± 3); there was no change in the heart rate between groups. At necropsy, we confirmed the effectiveness of the surgical fat ablation and found that BMX‐FR had a significant reduction in visceral fat expressed as % Body weight from 5.1± 0.5 to 1.0 ± 0.15. These data support the hypothesis that antenatal betamethasone exposure programs adipose tissue in a manner that leads to the impairment of cardiovascular system and that visceral fat removal early in life abrogate the cardiovascular dysregulation associated with betamethasone exposure. Further studies are being carried on to delineate the molecular changes in adipose tissues with fetal programming that leads to cardiometabolic disease and how it could be interrupted.

Support or Funding Information

HD‐47584, HL 68728
The Natriuretic and Diuretic Response to Uroguanylin is Blunted in Dahl‐Salt Sensitive Rats
Authors: Nascimento Nilberto Robson Falcão , Martins Isabel Cristina Mendonça Torres , Lessa Lucília Maria Abreu , Santos Cláudia Ferreira , Fonteles Manassés Claudino
Published: November 2023

Uroguanylin (UGN) is a 17 aminoacids long peptide with diuretic and natriuretic activity. UGN, an intestinal natriuretic peptide, is delivered to the kidney as an unprocessed propeptide, prouroguanylin (pro‐UGN). Circulating pro‐UGN is processed to its active natriuretic form exclusively within the renal tubules. The circulating levels of pro‐UGN and the urinary levels of UGN are increased during sodium overload and this natriuretic peptide and the kidney response to UGN is increased in rats maintained in a high salt intake. We hypothesized that the diuretic and natriuretic response to UGN are altered in Dahl‐salt sensitive rats (DS) as compared to Wistar (WR) rats. Dahl‐salt sensitive (DS) rats (n=5/group) received water with 1% NaCl during 15 days and the tail systolic blood pressure (mmHg) increased from 108.0±2.0 to 145.9±2.6 while no remarkable changes were observed in Wistar rats. After the end of this period, rats were anesthetized and prepared for classical renal clearance and hemodynamics studies. After a 1‐hour control period, UGN was intravenously infused at 100nmoles/kg/min rate during 1‐hour period and heart rate (HR), mean arterial pressure (MAP), renal vascular resistance (RVR), glomerular filtration rate (GFR), renal blood flow (RBF), urine output (V′), UNaV and fractional sodium excretion (FENa+) were evaluated during infusion and for an additional 1‐hour post‐infusion period. No significant changes were observed for HR, MAP, RVR or RBF in any group or period during the whole experimental period. UGN did not change GFR (ml/g/min) in WR rats but increased it from 1.7±0.8 to 4.4±1.1 in DS rats. In addition, UGN infusion induced a significant diuresis that was stable during the infusion and post‐infusion period. The urinary output (μL/g of kidney/min) increased from 6.7±1.1 to 21.6±5.4 (p<0.001) during the infusion with UGN in WR and from 5.7±2.6 to 11.5±3.6 in DS. Strikingly, the basal levels of UnaV (μEq/g of kidney/min) in DS are significantly lower than the levels observed for WR (0.04±0.01‐DS vs. 0.23±0.07‐p<0.001). The infusion of UGN increases UNaV from 0.23±0.07 to 0.48±0.01 (p<0.01) in WR and from 0.04±0.01 to 0.27±0.08 (P<0.001). Similarly, the basal FENa+ is remarkably lower in DS when compared to WR. DS rats excrete 10 times less sodium than WR. The infusion of UGN does not increase FENa+ (%) in DS but induced a 60% increase in in WR from 1.5±0.12 to 2.5±0.16 at the end of the infusion period. The diuretic and natriuretic effects of UGN are blunted in DS rats. The defect of this counter regulatory mechanism may have a role in the genesis and maintenance of hypertension in this strain of rats.

Support or Funding Information

CNPq, FUNCAP and CAPES
Diuretic and Natriuretic Activity of a Lipid Induced by Fatty Acyl Amide Hydrolase Inhibition in Cultured Mouse Medullary Interstitial Cells
Authors: Daneva Zdravka , Dempsey Sara , Ahmad Ashfaq , Wolf Carl , Li Ningjun , Li Pin‐Lan , Ritter Joseph
Published: November 2023

The endocannabinoid, anandamide (AEA, N‐arachidonoyl ethanolamine), is enriched in the renal medulla, a region implicated in the long‐term control of blood pressure. However, the role of AEA and its metabolizing enzymes, fatty acid amide hydrolase (FAAH) and cyclooxygenase‐2 (COX‐2), in the renal medulla are not well understood. In this study, cultures of mouse medullary interstitial cells (MMICs) from C57BL6 mice were established and characterized. Similar to inner medullary interstitial cells in mouse kidney sections in situ, cultured MMICs showed intense staining for tenascin C, a selective marker for medullary interstitial cells, and stained positively for immunoreactive COX‐2 and FAAH, and for the presence of lipid droplets by Oil Red O. Treatment of MMICs for 24 h with the FAAH inhibitor, PF3845, produced a marked increase in Oil Red O staining. Analysis of the chromatographic profiles of lipid extracts of the medium from vehicle‐ and PF3845‐treated MMIC cultures by reverse‐phase high pressure liquid chromatography (RP‐HPLC) with UV detection revealed a product that was responsive to treatment with PF3845. Levels of the product decreased when PF3845 was combined with the COX inhibitor, indomethacin. To assess the antihypertensive roles of the product, it was purified by RP‐HPLC of a larger scale extract and evaluated in anesthetized C57BL6 mice instrumented with a blood pressure probe in the carotid artery, a bladder cannula for collection of urine and an i.v. catheter implanted in the jugular vein for infusion of albumin‐saline. Continuous i.v. infusion of the lipid (0.1 AU/min for 20 min) had no effect on mean arterial blood pressure, but it increased urine volume and sodium and potassium excretion. The data suggest that the lipid is a diuretic‐natriuretic that is dependent on AEA or another ethanolamide and COX‐2 for its formation. Efforts are underway to identify the structure of the lipid.

Support or Funding Information

Supported by NIH grant DK 102539
Aquaporin‐3 deficiency slows renal cystogenesis via regulation of AMPK/ERK/mTOR signaling
Authors: Yang Baoxue , Wang Weiling
Published: November 2023

OBJECTIVE

Human autosomal dominant polycystic kidney disease (ADPKD) is characterized by numerous bilateral renal cysts that lead to a decline in kidney function over time. Previous studies reported aquaporin‐3 (AQP3) expression in cysts derived from collecting ducts in ADPKD. The aim of this study is to study the effect of AQP3 on ADPKD development.

METHODS

AQP3 deficient PKD mice and inducible PKD mice were established in this study. Effect of AQP3 on PKD development was studied with pathological and molecular biological techniques.

RESULTS

In both models, kidney size and cyst number were ~30% smaller in AQP3 null PKD mice than in AQP3‐expressing PKD mice, with the difference due mainly to smaller collecting ducts cysts. In matrix cultured MDCK cyst model, AQP3 transfection promoted cyst enlargement. To investigate the mechanism of AQP3‐dependent cyst development, we detected related signaling pathways. It was found that the AMPK/ERK/mTOR signaling, as well as intracellular ATP, were down‐regulated after AQP3 deletion. In addition, overexpression of AQP3 increased intracellular ATP synthesis, and HIF1α and glucose transporter 1 expression.

CONCLUSION

We conclude that AQP3 depletion retards cyst growth, primarily due to AQP3‐dependent energy metabolism. These findings identified the AQP3 as a potential therapeutic target for treating ADPKD.

Support or Funding Information

This work was supported by National Natural Science Foundation of China grants 31200869, 81261160507, 81330074 and 81170632, and the 111 Project.
Exogenous administration of hydrogen sulphide enhances the responsiveness of α1D‐adrenoreceptors in the kidney of rats with left ventricular hypertrophy
Authors: Ahmad Ashfaq , Sattar Munavvar Zubaid Abdul , Azam Maleeha , Khan Safia Akhtar , Abdullah Nor Azizan , John Edward James
Published: November 2023

Present study explored the interaction of hydrogen sulphide (H2S) and α1D‐adrenoreceptors of the kidney. This study investigated the functional contribution of α1D‐adrenoreceptors subtypes mediating the renal vasoconstriction in the kidney of rats with left ventricular hypertrophy treated with H2S.

Methodology

The LVH was induced by isoprenaline administration (5mg, S/C, every 72 hours for 2 weeks) and caffeine in drinking water (62mg/L) for 2 weeks while H2S was given 56μM/kg for 5 weeks as intra‐peritoneal injection. The responsiveness of α1D to exogenous noradrenaline (NA), phenylephrine (PE) and methoxamine (ME) in the absence and presence of BMY 7378 was studied.

Results

The plasma and kidney concentration of H2S in LVH group was significantly lower (P<0.05) than the control (LVH vs. control, 16±1 and 24±2 vs. 54±2 and 38±1 μM). The exogenous administration of NaHS in LVH resulted in a significant increased (P<0.05) in plasma and kidney concentration of H2S when compared to their untreated counterparts (LVH‐H2S vs. LVH, 54±2 and 67±2 vs. 16±1 and 24±2 μM). Induction of LVH significantly blunted (P<0.05) the responses of NA, PE and ME in saline phase of LVH by 21%, 23% and 28 % when compared to respective phase of Control group. Blocking of α1D‐adrenoceptor by low dose and high dose of BMY did not significantly reduced the responses to NA, PE and ME in low dose and high dose phases of LVH when compared to responses produced by NA, PE and ME in same phases of Control group. Exogenous administration of H2S significantly increased (P<0.05) the responses by α1D‐adrenoceptor to NA, PE and ME in LVH‐H2S in low and high dose phases by 52% and 30 %, 120% and 206 % while 116% and 423 % respectively when compared to responses produced by α1D‐adrenoceptor to NA, PE and ME in same phases of LVH

Conclusion

Treatment with H2S augmented the responsiveness of α1D‐adrenoceptor subtypes in the kidney of rats with LVH.

Support or Funding Information

The Institute of Postgraduate Studies (IPS) is acknowledged for the provision of a USM fellowship (Teaching) to Ashfaq Ahmad (APEX (1002/JHEA/ATSG4001). The authors fully acknowledge USM‐RU grant no. 1001/PFARMASI/815078 for this work.
PDI via Keap1/Nrf2 pathway Regulates Redox Balance, Renal AT1 Receptor Function and Blood Pressure
Authors: Wang Xitao , Asghar Mohammad
Published: November 2023

Oxidative stress is reported to play a vital role in the regulation of renal angiotensin AT1 receptor (AT1R) function and blood pressure (BP). This indicates that redox balance is crucial for normal AT1R function and BP. We tested a hypothesis that Protein Disulfide Isomerase (PDI), an enzyme that has diverse biological activity, via Keap1/Nrf2 pathway, regulates cellular redox homeostasis, AT1R function and blood pressure. To test the hypothesis, we treated adult Sprague Dawley (SD) rats with bacitracin, a PDI inhibitor for 14 days and determined various parameters. We found that bacitracin treated compared to vehicle treated rats, showed higher blood pressure. This was associated with greater diuresis and natriursis in response to AT1R blocker candesartan in bacitracin treated rats, indicating higher AT1R function in these rats. Moreover, bacitracin treated rats had higher oxidative stress and renal injury, as determined by measuring oxidative stress markers: urinary 8‐isoprostane, renal protein carbonyls and renal nitrotyrosine levels as well as urinary kidney injury molecule‐1 levels, respectively. This was associated with reduced antioxidant capacity and impaired Nrf2 transcription factor activity, involved in maintaining antioxidant capacity. Further, Kelch like ECH associated protein 1 (keap1) levels, an endogenous repressor of Nrf2, and tyrosine 216 phosphorylated GSK3β protein, which causes Nrf2 nuclear export were higher in bacitracin treated rats. Moreover, Nuclear Nrf2 levels in human kidney 2 cells treated with bacitracin also decreased compared to control cells. These data suggest that PDI via keap1/Nrf2 pathway regulates redox balance, AT1R function and BP in SD rats.

Support or Funding Information

NIH/NIA, AG039856
Characterization of potassium channels involved in the mechanisms of vascular relaxation induced by omega‐3 polyunsaturated fatty acids in rat arteries; a critical role for KATP channels
Authors: Limbu Roshan , Cottrell Graeme S , McNeish Alister J
Published: November 2023

Introduction

Fish oils are involved in the vasodilation of arteries and studies indicate that the main components involved are omega‐3 polyunsaturated fatty acids (n‐3 PUFA) that include docosahexaenoic acid (DHA) and eicosapentaenoic acids (EPA) (1). We previously demonstrated that inhibition of KCa channels partially attenuates fish oil mediated relaxation in rat mesenteric arteries (2). Since a considerable proportion of the relaxation to n‐3 PUFA was residual to blockade of KCa, vasodilation pathways involving other types of potassium channels could be involved. Studies report that the predominant vasodilation mechanisms can differ depending upon the type of artery an (3). Therefore, our main objective was to investigate these pathways in n‐3 PUFA‐induced relaxation of both large muscular (aorta) and resistance rat arteries (mesenteric).

Methods

2mm segments of rat arteries were mounted on a wire myograph and cumulative concentration response curves to DHA or EPA (10nM–30μM) mediated relaxation were examined following pre‐constriction with U46619 (10–500nM) in rat aorta and mesenteric artery. In certain experiments isotonic Krebs solution containing high [KCl] (final bath concentration 30mM; inhibits hyperpolarization mediated by potassium channels) or PNU37883A (3μM), a KATP blocker were administered prior to assessing relaxation. Relaxation was expressed as mean % relaxation of U46619‐induced constriction ± SEM of n animals. Data was analysed using one‐way ANOVA with Bonferroni's post‐test. P‐value of ≤ 0.05 was considered as being statistically significant.

Results

Non‐selective inhibition of potassium channels with high [KCl] Krebs virtually abolished both DHA‐ and EPA‐induced relaxation in rat aorta. Furthermore, High [KCl] Krebs also partially inhibited relaxation mediated by both DHA and EPA in rat mesenteric artery. Inhibition of KATP with PNU37883A attenuated fish oil‐induced relaxation in rat aorta. Consistently, in rat mesenteric artery PNU37883A also inhibited both DHA‐ and EPA‐induced relaxation.

Conclusions

We demonstrate that the non‐selective blockade of potassium channels abolished both DHA‐ and EPA‐mediated relaxation of rat aorta. Furthermore, inhibition of potassium channels in the resistance mesenteric artery also inhibited relaxation induced by DHA and EPA. We further characterised the potassium channels involved; blockade of KATP channels inhibited both DHA and EPA induced relaxation in rat aorta and mesenteric artery. This is the first study indicating that DHA and EPA evoke relaxation through KATP in both rat large muscular and resistance arteries. We will incorporate patch clamp in our future studies to investigate whether n‐3 PUFA directly regulate KATP channels.

Support or Funding Information

This work was supported by British Heart Foundation (PhD Studentship no. FS/14/33/30799)
Nstpbp5185 attenuates the progression of neointima formation in a rat model of balloon injury
Authors: Huang Tur‐Fu , Huang Shiu‐Wen
Published: November 2023

Background

Abnormal proliferation and migration of vascular smooth muscle cells (VSMCs) play an important role in the development of restenosis. Platelet‐derived growth factor (PDGF) is one of the major factors in triggering restenosis. Nstpbp5185 is a potent TP receptor antagonist, exhibits anti‐thrombotic activities.

Material and methods

In an effort to figure out the anti‐restenosis effect of nstpbp5185, we investigated its effects on VSMC proliferation, migration in vitro and neointima formation in rat balloon injury model in vivo.

Results

Our findings demonstrated that nstpbp5185 inhibited PDGF and U46619‐induced VSMC proliferation and PDGF‐induced migration in a concentration‐dependent manner. Furthermore, nstpbp5185 also concentration‐dependently repressed PDGF‐PDGFR signaling pathway in VSMCs. In addition, in vivo experiments showed that intraperitoneal administration of nstpbp5185 at 1 mg/kg/day significantly attenuated neointima formation in balloon‐injured rat carotid arteries.

Conclusions

Our results reveal that nstpbp5185 inhibits PDGF‐induced VSMC proliferation and migration, resulting in attenuation of neointima formation. Its therapeutic potential for treating restenosis after percutaneous transluminal coronary angioplasty (PTCA) warrants further evaluation.
Bazedoxifene Induces Greater Vascular Responses than Estradiol Independent of Sex and GPER
Authors: Zimmerman Margaret A. , Lindsey Sarah H.
Published: November 2023

Conjugated estrogens (CE) combined with the selective estrogen receptor modulator (SERM) bazedoxifene (BZA) is a novel menopausal hormone therapy that provides a progestin‐free option for women with an intact uterus. While the impact of estrogen (E2) on the vascular system has been extensively studied, less is known about the response to BZA. Previous work from our laboratory shows that many of the beneficial actions of estrogen in the vasculature are mediated by the G protein‐coupled estrogen receptor (GPER). Since previous generation SERMs bind to GPER in various tissues, we hypothesized that BZA may induce greater vascular effects than estradiol due to increased GPER activation. Mesenteric resistance arteries were isolated from male and female wildtype (WT) and GPER knockout (KO) mice between 18–24 weeks old. Vessel reactivity was measured via wire myography in response to increasing doses of phenylephrine (PE), sodium nitroprusside (SNP), bazedoxifene acetate (BZA), and 17β‐estradiol (E2). In males, BZA‐induced relaxation was significantly greater than E2 in both WT (BZA: 84 ± 7% vs. E2: 50 ± 5%; P<0.001) and KO vessels (BZA: 72 ± 11% vs. E2: 55 ± 4%; P<0.001). Similarly, BZA was a better vasodilator than E2 in females (BZA: 94 ± 1% vs. E2: 57 ± 6%; P<0.001), and was not altered by GPER deletion (BZA: 93 ± 1% and E2: 60 ± 6%; ns). To further assess the impact of BZA on vessel reactivity, mesenteric arteries were pre‐treated with either 10−9 M BZA, E2, or vehicle (0.1% DMSO) before being exposed to increasing doses of PE. In male WT and KO vessels, BZA blunted the response to PE‐induced contraction (WT Veh: 178 ± 11% vs. BZA: 145 ± 32%; P<0.001; KO Veh: 154 ± 5% vs. BZA: 120 ± 19%; P<0.001). This effect was exclusive to BZA and was not observed in male WT and KO vessels pre‐treated with E2 or DMSO (WT E2: 173 ± 20% and KO E2:160 ± 6%; ns). In female WT and KO arteries, pre‐treatment with BZA also opposed PE‐induced contraction (WT Veh: 159 ± 15% vs. BZA: 61 ± 10%; P<0.001; KO Veh: 173 ± 7% vs. BZA: 109 ± 20%; P<0.001), while E2 again failed to attenuate PE contraction (WT E2: 175 ± 20% and KO E2:189 ± 9%; ns). To assess whether BZA enhances nitric oxide‐induced relaxation, vessels were pre‐treated with BZA, E2, or DMSO prior to the SNP relaxation curve. Neither E2 nor BZA altered the response to SNP in male or female, WT or KO vessels. The current study demonstrates that BZA opposes vasoconstriction of mesenteric arteries to a greater degree than non‐selective E2, and this effect is independent of sex and GPER. Therefore, BZA may provide additional vascular benefits in menopausal hormone therapy and in turn promote a more positive cardiovascular profile in postmenopausal women.

Support or Funding Information

National Institutes of Health Grant 4R00HL103974 to S.H.L and American Heart Association Postdoctoral Fellowship Award #16POST27600001 to M.A.Z.