Newborn Rats Research Articles

Differentiation of primary retinal progenitor cells into retinal ganglion-like cells using low dose cytarabine.

The death of retinal ganglion cells (RGCs) is a key factor in the pathophysiology of all forms of glaucoma. RGC culture serves as a simple system for establishing and testing candidate therapies. This study aimed to explore the differentiation of primary retinal progenitor cells (RPCs) into RGC-like cells induced by low-dose cytarabine (Ara-C). RPCs were isolated from the retina of newborn rats and cultured in vitro. Different concentrations of Ara-C were added to the culture medium to induce the differentiation of RPCs into RGC-like cells. Differentiation efficiency was assessed through immunofluorescence staining and cell counting. The addition of Ara-C significantly increased the number of Brn3a/RBPMS double-positive cells. The RPC-RGCs induced displayed characteristic features of RGCs, with roughly 80.9%±6.2% of the cells positive for both TuJ1/NeuN and 77.5%±4.9% for Brn3a/RBPMS. The study demonstrates that the addition of Ara-C to primary cultures of rat RPCs can enhance their differentiation into RGC-like cells, providing a simple and rapid method for obtaining RGC-like cells with a relatively high purity. This method shows considerable promise for advancing glaucoma research and potential therapeutic strategies to restore vision after RGC loss.

Potassium channel‐mediated NO‐induced vasodilation during maturation: Dominance of Kv7 channels

AbstractMaturation represents a process characterized by adaptive changes, particularly in the circulatory system. However, it is not known whether, in neonates, potassium channels contribute to NO‐induced vasorelaxation at all and, if so, which potassium channels these are. Therefore, this study tested the hypothesis that potassium channels mediate NO‐induced vasorelaxation in newborn rats. Young (10‐ to 15‐day‐old) and adult (2‐ to 3‐month‐old) male rats were studied using real‐time PCR, isometric myography, and the sharp microelectrode technique on saphenous arteries. We observed prominent mRNA expression of several distinct isoforms of potassium channel families known to potentially mediate SNP‐induced vasodilation. Further, in both adult and young rats, SNP can relax vessels independently of potassium channels. A solely potassium channel‐independent anticontractile effect of SNP was observed also when either Kir6, or Kir2, or Kv2 channels, respectively, were available in both adult and young rats. However, when Kv1 channels were available, a Kv1 channel‐dependent component contributed to the anticontractile effect of SNP in young rats. When BKCa channels were available, a BKCa channel‐dependent component contributed to the anticontractile effect of SNP in adult rats. A considerable Kv7 channel‐dependent component contributed to the anticontractile effect of SNP in both adult and young rats. Thus, the data of the present study show for the first time that potassium channels, even multiple ones, contribute to SNP‐induced vasorelaxation in newborn rats and that the potassium channels involved in SNP‐induced vasorelaxation change from Kv1/Kv7 channels to BKCa/Kv7 channels during postnatal development.

Physiologic and structural characterization of desisobutyryl-ciclesonide, a selective glucocorticoid receptor modulator in newborn rats.

Bronchopulmonary dysplasia, the most prevalent chronic lung disease of prematurity, is often treated with glucocorticoids (GCs) such as dexamethasone (DEX), but their use is encumbered with several adverse somatic, metabolic, and neurologic effects. We previously reported that systemic delivery of the GC prodrug ciclesonide (CIC) in neonatal rats activated glucocorticoid receptor (GR) transcriptional responses in lung but did not trigger multiple adverse effects caused by DEX. To determine whether limited systemic metabolism of CIC was solely responsible for its enhanced safety profile, we treated neonatal rats with its active metabolite desisobutyryl-ciclesonide (Des-CIC). DEX but not Des-CIC caused a reduction in body weight as well as reduced insulin-like growth factor-1 serum levels and chronic hyperglycemia in neonatal rats. However, Des-CIC was as effective as DEX in reducing the expression of various bleomycin-induced proinflammatory cytokine mRNAs. In vitro studies with various cell types demonstrate the potent GR transactivation and transrepression activity of Des-CIC, although genome-wide transcriptomic analyses reveal differences in DEX vs. Des-CIC responses in neonatal rat lung and liver tissue. Des-CIC is a GR super-agonist as revealed by an in vitro coregulator peptide binding assay. In addition, molecular dynamics simulations revealed unique Des-CIC-dependent allosteric signaling pathways between specific residues in the GR ligand-binding domain and receptor surfaces interacting with coregulator peptides. Thus, Des-CIC is a potential novel selective GR modulator that could impart a favorable therapeutic index for CIC use for even modest durations of GC exposure which could have long-lasting adverse somatic, metabolic, or neurologic effects.

Investigation of the Effect of Wheat and Corn Gluten on Inflammation, Transglutaminase, Gliadin and IgA Levels in Healthy Rat Intestines

The aim of this study was to evaluate the effects of wheat and corn gluten on some histopathologic parameters such as villus atrophy, crypt hyperplasia, lymphocyte plasma neutrophils and immunohistochemical parameters such as trans glutaminase, gliadin and IgA in the small intestine of healthy male rats without HLA-DQ2 and HLA-DQ8 genes. In the study, 21 healthy newborn male Sprague Dawley rats were fed wheat, corn and soy with the addition of 7 rats in each group from one-day age to 60 days of age. Histopathological (villous atrophy, lymphocyte plasma neutrophil, crypt hyperplasia) and immunohistochemical (transglutaminase, gliadin, IgA) parameter analyses were performed in small intestinal tissue samples. As a result of the study, it was found that the small intestinal villus lengths of the wheat gluten group were longer than the other groups (P < .05). Cryptic hyperplasia was detected most in the soybean group and the lowest in the wheat group (P < .05). Gliadin antibody levels were found to be in the soybean group with the highest and the lowest in the wheat group (P < .05). In healthy male rats lacking HLA-DQ2 and HLA-DQ8 genes, the effect of wheat gluten on crypt hyperplasia and gliadin levels in small intestinal tissue was significantly lower than in soy and corn gluten groups, while its effect on villous atrophy, lymphocyte plasma neutrophil and transglutaminase was limited. In addition, the intestinal villus lengths of the wheat gluten group were significantly higher than those of the corn and soybean groups.

Increased ROS levels, antioxidant defense disturbances and bioenergetic disruption induced by thiosulfate administration in the brain of neonatal rats.

Sulfite oxidase deficiencies, either caused by deficiency of the apoenzyme or the molybdenum cofactor, and ethylmalonic encephalopathy are inherited disorders that impact sulfur metabolism. These patients present with severe neurodeterioration accompanied by cerebral cortex and cerebellum abnormalities, and high thiosulfate levels in plasma and tissues, including the brain. We aimed to clarify the mechanisms of such abnormalities, so we assessed the ex vivo effects of thiosulfate administration on energetic status and oxidative stress markers in cortical and cerebellar tissues of newborn rats. Thiosulfate (0.5 µmol/g) or PBS (vehicle) was injected into the fourth ventricle of rat pups. Thirty minutes after the injection, animals were euthanized and the brain structures were utilized for the experiments. Our data showed that thiosulfate decreased the reduced glutathione (GSH) concentrations, and superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) activities in the cortical structure. Thiosulfate also increased DCFH oxidation, hydrogen peroxide generation and glutathione reductase activity. In the cerebellum, thiosulfate reduced SOD and glutathione peroxidase activities but increased GST and CAT activities as well as DCFH oxidation. Regarding energy metabolism, thiosulfate specifically decreased complex IV activity in the cortex, whereas it increased cerebellar complex I and creatine kinase activities, indicating bioenergetic disturbances. The results suggest that the accumulation of thiosulfate causing redox disruption and bioenergetic alterations has a prominent role in the pathogenesis of sulfur metabolism deficiencies.

One-Week Maternal Separation Caused Sex-Specific Changes in Behavior and Hippocampal Metabolomics of Offspring Rats.

To investigate the effects of one-week maternal separation (MS) on anxiety- and depression-like behaviors in adolescent and adulthood as well as adult hippocampal metabolomics simultaneously in offspring female and male rats. In the MS group, newborn SD rats were separated from their mothers for 3 h per day from postnatal days (PND) 2 to 8. The open field test (OFT), elevated plus mazes (EPM), novelty suppressed feeding test (NSFT), and forced swimming test (FST) were conducted during adolescence and adulthood. Serum corticosterone, mRNA expression of hippocampal inflammatory cytokines, and hippocampal untargeted metabolomics of offspring adult rats were examined using an assay kit, qRT-PCR, and UPLC-Q-TOF/MS. Both MS female and male rats showed similar behaviors in OFT, EPM, NSFT, and SPT, except for the latency to feeding during adolescence and the open arm entries during adulthood, showed statistical significance only in MS female rats. Serum corticosterone and hippocampal pro-inflammatory cytokines IFN-γ were significantly elevated in both female and male rats, and IL-1β and TNF-α were significantly increased only in female rats. In hippocampal metabolism, the identification of differential metabolites displayed 53 and 37 in female rats and male rats, respectively (with 35 common metabolites), which were involved in 33 and 30 metabolic pathways with 28 common pathways. One-week MS induced sex-specific anxiety- and depression-like behaviors in female and male offspring rats during adolescence and adulthood, as well as sex-differentiated characteristics in the hippocampus inflammatory cytokines and metabolomics of adult MS rats. From the experimental data, the effects of MS on the female offspring rats were more severe than those of the male offspring rats.

Participation of adrenoreceptors in the mechanisms of pathologic cardiac rhythm induced in newborn rats by nickel chloride administration

Participation of adrenoreceptors in the mechanisms of pathologic cardiac rhythm induced in newborn rats by nickel chloride administration

Comparative Analysis of the Effects of Maternal Hypoxia and Placental Ischemia on HIF1-Dependent Metabolism and the Glucocorticoid System in the Embryonic and Newborn Rat Brain

Comparative Analysis of the Effects of Maternal Hypoxia and Placental Ischemia on HIF1-Dependent Metabolism and the Glucocorticoid System in the Embryonic and Newborn Rat Brain

Effect of Extrauterine Growth Restriction and Subsequent Catch-Up Growth on Cardiac Parameters: A Study on Wistar Rat Pups.

Objective: To study the long-term effect of postnatal growth restriction followed by catch-up growth on microscopic cardiac parameters in newborn Wistar rats. Methodology: This lab-based study was conducted at the Anatomy Department, CIMS Multan, on 120 neonate Wistar rats utilizing a rodent litter size manipulation model which mimicked human extrauterine growth restriction (EUGR) and catch-up growth. Initially, 30 pups were fed normally (Group N), while 90 were undernourished (Group R) to simulate EUGR. From days 11 to 21, the undernourished rats were separated into two subgroups: normal growth (RN, n = 30) and catch-up growth (RC, n = 60). The RC subgroup was further subdivided into accelerated (RCA) and slow (RCS) catch-up growth based on growth velocity. On day 60, microscopic cardiac parameters were assessed after euthanasia by ANOVA with a post-hoc Tukey test. Results: Malnourished rats with catch-up growth (RCS and RCA) had significantly increased left ventricle wall area, decreased left ventricle lumen area and a higher mean cardiomyocyte diameter than that of the normally fed (N) and malnourished than normally fed (RN) rats. Also, RCA pups had significantly thicker left ventricular walls and less lumen area than RCS pups. No difference in the collagen content of myocardium was detected among the subgroups. Conclusion: Accelerated and slow catch-up growth in rat pups decreases the left ventricular lumen area and increases the left ventricular wall + interventricular septum area significantly. Also, it increases the cardiomyocyte diameter in cardiac tissue.

Altered purine and pentose phosphate pathway metabolism in uteroplacental insufficiency-induced intrauterine growth restriction offspring rats impairs intestinal function

BackgroundThis study aimed to identify metabolic alterations in the small intestine of newborn rats with intrauterine growth restriction (IUGR), a condition linked to intestinal dysfunction. MethodsPregnant Sprague Dawley rats underwent bilateral uterine artery ligation on gestational day 17 to induce intrauterine growth restriction or sham surgery. Rat pups were delivered spontaneously on gestational day 22. Small intestine tissues were collected on postnatal days 0 and 7 from offspring. Liquid chromatography-mass spectrometry analysis was performed to investigate untargeted metabolomic profiles. Western blot analysis assessed protein expression of key regulators. ResultsNewborn rats with intrauterine growth restriction exhibited distinct small intestine metabolic profiles compared to controls on postnatal day 0. Notably, significant alterations were observed in purine metabolism, the pentose phosphate pathway, and related pathways. Western blot analysis revealed a decrease expression in transketolase, a key enzyme of the pentose phosphate pathway, suggesting impaired activity of the pentose phosphate pathway. Additionally, decreased expression of tight junction proteins ZO-1 and occludin indicated compromised intestinal barrier function in rats with intrauterine growth restriction. Similar metabolic disruptions persisted on postnatal day 7, with further reductions in tricarboxylic acid cycle intermediates and folate biosynthesis precursors. Interestingly, lysyl-glycine, a protein synthesis marker, was elevated in rats with intrauterine growth restriction. ConclusionsOur findings reveal a distinct metabolic signature in the small intestine of neonatal rats with intrauterine growth restriction, characterized by disruptions in the pentose phosphate pathway, purine metabolism, and energy production pathways. These novel insights suggest potential mechanisms underlying IUGR-associated intestinal dysfunction and impaired growth.

Single-Cell RNA Sequencing to Guide Autologous Preterm Cord Mesenchymal Stromal Cell-Therapy.

The chronic lung disease bronchopulmonary dysplasia (BPD) remains the most common complication of extreme prematurity (<28 weeks of gestation). Umbilical cord-derived mesenchymal stromal cells (UC-MSCs) represent an opportunity for autologous cell-therapy, as UC-MSCs have been shown to improve lung function and structure in experimental BPD. However, characterization and repair capacity of UC-MSCs derived from donors with pregnancy-related complications associated with prematurity remain unexplored. To characterize UC-MSCs' transcriptome and determine if pregnancy-related complications (preeclampsia and chorioamnionitis) alter their therapeutic potential. Single-cell RNA sequencing (scRNA-seq) was used to compare the transcriptome of UC-MSCs derived from five term donors, 16 preterm donors, and human neonatal dermal fibroblasts (HNDFs, control cells of mesenchymal origin), and correlated with their therapeutic potential in experimental BPD. Using publicly available neonatal lung single-nuclei RNA sequencing data, we also determined putative communication networks between UC-MSCs and resident lung cell populations. Most UC-MSCs displayed a similar transcriptome despite of their pregnancy-related conditions and mitigated hyperoxia-induced lung injury in newborn rats. Conversely, HNDFs, one term and two preeclampsia preterm UC-MSC donors exhibited a distinct transcriptome enriched in genes related to fibroblast function and senescence and were devoid of therapeutic benefit in hyperoxia-induced BPD. Conversely, therapeutic UC-MSCs displayed a unique transcriptome active in cell proliferation and distinct cell-cell interactions with neonatal lung cell populations, including NEGR and NRNX pathways. Term and preterm UC-MSCs are lung protective in experimental BPD. scRNA-seq allows to identify donors with a distinct UC-MSC transcriptome characteristic of reduced therapeutic potential.

Visseral Lipectomy Improves Metabolic Syndrome Parameters and Adipokines in a Rat Model of Metabolic Syndrome Induced by Monosodium Glutamate.

Metabolic syndrome (MetS) includes abdominal obesity, hypertension, insulin resistance, and dyslipidemia. Research has indicated that reducing excess visceral fat has positive effects on inflammation and insulin resistance. We examined whether visceral lipectomy modifies the effects of MetS parameters and adipocytokine levels. Each group included 15 newborn male rats: control+sham (C+S), metabolic syndrome+sham (MetS+S), and metabolic syndrome+visceral lipectomy (MetS+VL). On postnatal days 0, 2, 4, 6, 8, and 10, subcutaneous injections of monosodium glutamate (MSG) (4g/mg) were administered to induce MetS. The control group received saline injection. The rats underwent sham surgery or lipectomy on the 120th day of life. Two months post-surgery, tests were performed to check lipid and insulin levels as well as the Lee index, HOMA-IR, serum adiponectin (ADP), resistin, interleukin-6 (IL6), leptin, tumor necrosis factor-alpha (TNF-α), and C-reactive protein (CRP) levels. These findings showed that the Lee index (p = 0.001), insulin resistance (p = 0.002), and hyperinsulinemia (p = 0.009) were significantly improved in the MetS+VL group compared to those in the MetS+S group. The lipid profile was unaffected by visceral lipectomy. Furthermore, visceral lipectomy normalized MetS-induced adipokine imbalance. The decrease in the Lee index and improvement in hyperinsulinemia suggest that visceral lipectomy may benefit impaired glucose metabolism. Although visceral lipectomy has no apparent effect on the lipid profile, positive effects on adipokine levels by reducing various inflammatory markers including resistin, IL6, leptin, TNF-α, and CRP. These findings indicate that visceral lipectomy may have therapeutic potential for MetS. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Piezo1 Regulates Stiffness-Dependent DRG Axon Regeneration via Modifying Cytoskeletal Dynamics.

Despite medical interventions, the regenerative capacity of the peripheral nervous system is limited. Dorsal root ganglion (DRG) neurons possess the capacity to detect mechanical signals from their microenvironment, but the impact and mechanism by which these signals regulate axon regrowth and even regeneration in DRG neurons remain unclear. In this study, DRG neurons from newborn rats are cultured on substrates with varying degrees of stiffness in vitro to investigate the role of mechanical signals in axon regrowth. The findings reveal that substrate stiffness plays a crucial role in regulating axon regrowth, with an optimal stiffness required for this process. In addition, the data demonstrate that Piezo1, a mechanosensitive cation channel, detects substrate stiffness at the growth cone and regulates axon regrowth through activating downstream Ca2+-CaMKII-FAK-actin cascade signaling pathway. Interestingly, knocking down Piezo1 in adult rat DRG neurons leads to enhanced axon regeneration and accelerated recovery of sensory function after sciatic nerve injury. Overall, these findings contribute to the understanding of the role of mechanical signals in axon regeneration and highlight microenvironmental stiffness as a promising therapeutic target for repairing nerve injuries.

Lactobacillus rhamnosus modulates murine neonatal gut microbiota and inflammation caused by pathogenic Escherichia coli

BackgroundPathogenic Escherichia coli strains produce neonatal septicemia after colonizing the neonatal gut. While the probiotic Lactobacillus rhamnosus GG (LGG) effectively reduces neonatal sepsis, LGG’s effects on the neonatal intestinal microbiota alterations and inflammation triggered by E. coli are incompletely understood. We hypothesized that LGG significantly modulates the specific neonatal gut microbial populations changes and the inflammatory response elicited by the enteral introduction of septicemia-producing E. coli. To test this hypothesis, newborn rats were pretreated orally with LGG or placebo prior to infection with the neonatal E. coli septicemia clinical isolate SCB34. Amplicon 16S rRNA gene sequencing was performed on intestinal samples. Intestinal injury and expression of inflammatory mediators and apoptosis were determined.ResultsAlpha diversity of gut microbiota was greater in SCB34-infected pups in comparison to sham-infected pups, these changes were not modified by LGG pretreatment. Beta diversity analyses also showed differences between SCB34-infected vs. uninfected pups. LGG pretreatment before SCB34 infection did not result in significant beta diversity changes compared to placebo. Moreover, individual genera and species abundance analyses by linear discriminant analysis effect size (LEfSe) showed significant changes in Gram-negative, Gram-positive, and anaerobic populations resulting from LGG pretreatment and SCB34 infection. LGG significantly suppressed the expression of inflammatory cytokines but did not attenuate SCB34-induced apoptosis or histologic injury.ConclusionsLGG modulates clinically significant microbiota features and inflammation triggered by pathogenic E. coli intestinal infection shortly after birth. This new knowledge can potentially be harnessed to design novel interventions against gut-derived neonatal sepsis.

The protective effect of DMI on hippocampus EEG, behavioral and biochemical parameters in hypoxia-induced seizure on neonatal period.

Hypoxia-Induced Neonatal Seizure (HINS) is a prevalent type of seizure in infants caused by hypoxic conditions, which can lead to an increased risk of epilepsy, learning disabilities, and cognitive impairments later in life. This study focuses on examining the effects of dimethyl itaconate (DMI) on cognition, motor coordination, and anxiety-like behavior in male rats that have experienced HINS. 42 male Wistar newborn rats (PND10) were randomly divided into six groups (n = 7). 1) Control (Vehicle only); received DMI solvent (0.1ml) without applying hypoxia. 2-3) DMI; receiving (20 and 50 mg/kg; i.p). 4) HINS; they were placed in a hypoxia chamber with 7% oxygen and 93% nitrogen concentration for 15 minutes. 5-6) DMI+HINS; received DMI (20 and 50 mg/kg; i.p) 24h before hypoxia. Behavioral tests including; Novel object recognition test, Rotarod, Parallel bar, Open field and elevated plus maze (EPM); started at age 45 after birth. After behavioral tests, the hippocampal CA1 region local EEG was recorded in all groups. Then the brain hippocampus tissue was isolated and the amount of MDA, SOD, NO, and Thiol was measured by ELISA method. Data showed that the administration of DMI improved motor symptoms, anxiety-like behaviors, and cognition in HINS rats (p<0.05). EEG power in the HINS group decreased significantly compared to other experimental groups (p<0.05). Biochemical observations showed that DMI significantly reduced oxidative stress and inflammation in the hippocampal tissue of HINS rats (p<0.05). Increased hippocampal oxidative stress and inflammation can be effective in the occurrence of behavioral disorders observed in HINS rats. While DMI improved these behavioral impairments by reducing oxidative stress and inflammation.

ИЗУЧЕНИЕ МИТОХОНДРИАЛЬНОЙ ТОКСИЧНОСТИ ПАРАЦЕТАМОЛА И ЕГО МЕТАБОЛИТА NAPQI

Introduction. Antipyretic effect of paracetamol has been known since the late 19th century. But the safety profile of this drug started to be analyzed much later, when some toxic effects were identified in using a drug that contained paracetamol, the most widely known of which was drug-induced liver damage. Aim. To study the effects of paracetamol and its toxic metabolite N-acetyl-p-benzoquinone-imine on the viability and mitochondrial membrane potential of astroglial cells in the cerebral cortex of newborn rats. Materials and Methods. Isolated astrocytes from the brains of newborn rats were exposed to 1 mM of paracetamol or 0.15 mM of its toxic metabolite NAPQI for 24 hours, then cell viability (colorimetric test for assessing the metabolic activity of cells, i.e. MTT method) and mitochondrial membrane potential (using TMPE fluorochrome) were identified. Results and Discussion. The paracetamol metabolite NAPQI decreased cell viability from 0.57 ± 0.02 down to 0.4507 ± 0.03 a.u. and mitochondrial membrane potential from 0.19±0.01 down to 0.16±0.003 a.u., the differences are significant compared to the control (p &lt;0.05). Conclusions. The results obtained indicate the toxic effect provided by the paracetamol metabolite NAPQI on astroglial cells in the brains of newborn rats. Keywords: paracetamol, NAPQI, mitochondrial toxicity, astroglial cells.

Prenatal exposure to di-n-butyl phthalate promotes RIPK1-regulated necroptosis of uroepithelial cells and induces hypospadias through the epithelial-mesenchymal transition process in newborn male rats

Maternal exposure to di-n-butyl phthalate (DBP) has been linked to the induction of hypospadias; however, the underlying mechanism remains unclear. Necroptosis is reported to be implicated in developmental malformations. This study aimed to investigate the underlying mechanism of necroptosis in the development of hypospadias. DBP was dissolved in corn oil, and pregnant rats were administered a precisely measured dose of DBP (750 mg/kg/day) via gastric intubation from gestation day 14–18. Control rats received only corn oil. The day of birth was considered postnatal day (PND) 1. Male hypospadias rats were identified on PND 7. Genital tubercle tissues were collected and stored at −80°C for subsequent PCR analysis, cryopreserved in liquid nitrogen for western blot, or fixed in formalin for immunohistochemistry (IHC) staining. IHC staining and western blot analysis revealed increased expression of RIPK1 and necroptosis markers in genital tubercle (GT) tissue compared to the control group. Additionally, higher levels of EMT and impaired androgen receptor expression were observed in GT tissue. Exposure to increased DBP concentrations in rat primary uroepithelial cells (PUCs) led to elevated ROS production. Necroptosis markers and EMT expression levels were upregulated in PUCs following DBP incubation. Notably, treatment with DBP combined with necrostatin-1, a necroptosis inhibitor, reduced the expression of EMT markers and ROS production compared to DBP treatment alone. In vitro studies further revealed that DBP-induced necroptosis promoted the degradation of E-cadherin through the ubiquitin-proteasome pathway in PUCs. Our findings suggest that maternal exposure to DBP promotes necroptosis in uroepithelial cells by elevating ROS level and EMT status. Thus, necroptosis may play an essential role in the development of hypospadias.

MiR-497-5p Ameliorates Deep Venous Thrombosis by Facilitating Endothelial Progenitor Cell Migration and Angiogenesis by Regulating LITAF.

Deep vein thrombosis (DVT) is a clinical manifestation of venous thromboembolism and a major global burden of cardiovascular disease. In recent years, the crucial role of microRNAs (miRNAs) in cardiovascular disease has been confirmed. Here, we aimed to investigate the specific effect of miR-497-5p on DVT. The endothelial progenitor cells (EPCs) were obtained from the bone marrow of newborn rats and transfected with miR-497-5p mimics or/and pcDNA3.1/lipopolysaccharide-induced TNF factor (LITAF). The proliferation and migration abilities of EPCs were detected using CCK-8 assay and transwell assay, respectively. Angiogenesis was evaluated using tube formation assay. The interaction of miR-497-5p and LITAF was confirmed by luciferase reporter experiment. DVT rat model in vivo was established by inferior vena cava (IVC) ligation in Sprague-Dawley rats. Histological analysis of IVC tissue was conducted by hematoxylin-eosin staining. We found that enhancing miR-497-5p expression facilitated the abilities of proliferation and migration of EPCs. Additionally, overexpression of miR-497-5p increased the capacity of EPCs to form capillary tubes on Matrigel. LITAF was found to be targeted by miR-497-5p and negatively regulated by miR-497-5p. Overexpression of LITAF counteracted the miR-497-5p overexpression's effect on the proliferation, migration, and angiogenesis abilities of EPCs. Moreover, the injection of agomir-miR-497-5p alleviated thrombus formation, reduced thrombus weight, and reduced the serum level of D-dimer in DVT rat model by reducing LITAF expression. This study suggests that miR-497-5p alleviates DVT by facilitating EPCs proliferation, migration, and angiogenesis by targeting LITAF.

Nintedanib preserves lung growth and prevents pulmonary hypertension in a hyperoxia-induced lung injury model.

Bronchopulmonary dysplasia (BPD), the chronic lung disease associated with prematurity, is characterized by poor alveolar and vascular growth, interstitial fibrosis, and pulmonary hypertension (PH). Although multifactorial in origin, the pathophysiology of BPD is partly attributed to hyperoxia-induced postnatal injury, resulting in lung fibrosis. Recent work has shown that anti-fibrotic agents, including Nintedanib (NTD), can preserve lung function in adults with idiopathic pulmonary fibrosis. However, NTD is a non-specific tyrosine kinase receptor inhibitor that can potentially have adverse effects on the developing lung, and whether NTD treatment can prevent or worsen risk for BPD and PH is unknown. We hypothesize that NTD treatment will preserve lung growth and function and prevent PH in an experimental model of hyperoxia-induced BPD in rats. Newborn rats were exposed to either hyperoxia (90%) or room air (RA) conditions and received daily treatment of NTD or saline (control) by intraperitoneal (IP) injections (1 mg/kg) for 14 days, beginning on postnatal day 1. At day 14, lung mechanics were measured prior to harvesting lung and cardiac tissue. Lung mechanics, including total respiratory resistance and compliance, were measured using a flexiVent system. Lung tissue was evaluated for radial alveolar counts (RAC), mean linear intercept (MLI), pulmonary vessel density (PVD), and pulmonary vessel wall thickness (PVWT). Right ventricular hypertrophy (RVH) was quantified with cardiac weights using Fulton's index (ratio of right ventricle to the left ventricle plus septum). When compared with RA controls, hyperoxia exposure reduced RAC by 64% (p < 0.01) and PVD by 65% (p < 0.01) and increased MLI by 108% (p < 0.01) and RVH by 118% (p < 0.01). Hyperoxia increased total respiratory resistance by 94% and reduced lung compliance by 75% (p < 0.01 for each). NTD administration restored RAC, MLI, RVH, PVWT and total respiratory resistance to control values and improved PVD and total lung compliance in the hyperoxia-exposed rats. NTD treatment of control animals did not have adverse effects on lung structure or function at 1 mg/kg. When administered at higher doses of 50 mg/kg, NTD significantly reduced alveolar growth in RA controls, suggesting dose-related effects on normal lung structure. We found that NTD treatment preserved lung alveolar and vascular growth, improved lung function, and reduced RVH in experimental BPD in infant rats without apparent adverse effects in control animals. We speculate that although potentially harmful at high doses, NTD may provide a novel therapeutic strategy for prevention of BPD and PH. Anti-fibrotic therapies may be a novel therapeutic strategy for the treatment or prevention of BPD. High-dose anti-fibrotics may have adverse effects on developing lungs, while low-dose anti-fibrotics may treat or prevent BPD. There is very little preclinical and clinical data on the use of anti-fibrotics in the developing lung. Dose timing and duration of anti-fibrotic therapies may be critical for the treatment of neonatal lung disease. Currently, strategies for the prevention and treatment of BPD are lacking, especially in the context of lung fibrosis, so this research has major clinical applicability.

The effect of prenatal fumonisin B exposure on bone innervation in newborn Wistar rats.

This study explored the effects of prenatal exposure to fumonisins B (FB) on bone innervation in newborn Wistar rats. Pregnant dams (n = 6 per group) were assigned to either the control or one of two FB-exposed groups (60 mg or 90 mg/kg body weight) from the 7th day of gestation until parturition. On the day of parturition, one male pup from each litter (n = 6 per group) was randomly selected and euthanised, and their femurs were dissected for analysis. Bone innervation was quantified by examining the morphology patterns of sympathetic, parasympathetic, sensory and cocaine- and amphetamine-regulated transcript (CART)-positive fibres. Prepared bone sections were analysed using immunohistochemistry staining for protein gene product 9.5, tyrosine hydroxylase, choline acetyltransferase, vasoactive intestinal peptide (VIP), substance P and CART-positive neurons. The group that received a higher dose of FB demonstrated an increase in both the size and complexity of the complete bone neuronal network together with heightened sympathetic and sensory innervation, and displayed a decrease in neuron density and sympathetic innervation. Fumonisin B exposure led to a decrease in galanin-positive and VIP-positive bone neuronal networks in both groups exposed to FB, while in the lower-dose group, there was also a decrease in CART-positive innervation. Prenatal FB exposure significantly influences the neuronal bone network of rats, which is essential for maintaining bone homeostasis. These findings emphasise the necessity for further research to understand the lasting effects and underlying mechanisms of alterations induced by FB.