RESUMEN

Osteosarcoma is the most prevalent form of primary malignant bone tumor, primarily affecting children and adolescents. The nerve growth factors (NGF) referred to as neurotrophins have been associated with cancer-induced bone pain; however, the role of NGF in osteosarcoma has yet to be elucidated. In osteosarcoma samples from the Genomic Data Commons data portal, we detected higher levels of NGF and M2 macrophage markers, but not M1 macrophage markers. In cellular experiments, NGF-stimulated osteosarcoma conditional medium was shown to facilitate macrophage polarization from the M0 to the M2 phenotype. NGF also enhanced VCAM-1-dependent monocyte adhesion within the osteosarcoma microenvironment by down-regulating miR-513c-5p levels through the FAK and c-Src cascades. In in vivo xenograft models, the overexpression of NGF was shown to enhance tumor growth, while the oral administration of the TrK inhibitor larotrectinib markedly antagonized NGF-promoted M2 macrophage expression and tumor progression. These results suggest that larotrectinib could potentially be used as a therapeutic agent aimed at mitigating NGF-mediated osteosarcoma progression.

Asunto(s)

Monocitos , Factor de Crecimiento Nervioso , Osteosarcoma , Microambiente Tumoral , Molécula 1 de Adhesión Celular Vascular , Osteosarcoma/metabolismo , Osteosarcoma/tratamiento farmacológico , Osteosarcoma/patología , Humanos , Factor de Crecimiento Nervioso/metabolismo , Animales , Microambiente Tumoral/efectos de los fármacos , Monocitos/metabolismo , Monocitos/efectos de los fármacos , Molécula 1 de Adhesión Celular Vascular/metabolismo , Ratones , Adhesión Celular/efectos de los fármacos , Línea Celular Tumoral , Neoplasias Óseas/metabolismo , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/patología , Macrófagos/metabolismo , Pirimidinas/farmacología , Pirimidinas/uso terapéutico , Pirazoles/farmacología , Pirazoles/uso terapéutico , Ratones Desnudos

RESUMEN

INTRODUCTION: Ischaemic stroke induces oxidative stress, mitochondrial damage, inflammation and senescence and the decrease of cognitive function. Vitamin D is a fat-soluble vitamin that has a neuroprotective effect to repair the function of the nervous system. The aim of this study is to investigate the effect of vitamin D on memory function, p16, p21 (senescence), and nerve growth factor (NGF) mRNA expression on the hippocampus after transient global cerebral ischemic. MATERIALS AND METHODS: The study was designed as quasiexperimental with a control group that only received posttests. We performed in vivo study with an induction bilateral common carotid artery occlusion (BCCAO) model and vitamin D injection for 10 days. A total of 24 rats were divided into four groups (n = 6): Sham operation (SO [control]), BCCAO (transient global cerebral ischemic model not given vitamin D), VD1 (BCCAO + vitamin D 0.125 µg/kgBW), and VD2 (BCCAO + vitamin D 0.5 µg/kgBW). The spatial memory function was tested with the Morris water maze. We performed immunohistochemistry to localise p16 expression. p16, p21 and NGF mRNA expression were assessed by reverse transcriptase (RT-PCR) method. RESULTS: The vitamin D treatment group required shorter mileage to find the platform and probe test. The total time spent was longer in the target quadrant than in non-target. The Vitamin D-treated group had lower p16 and p21 mRNA expression and higher NGF mRNA expression than the BCCAO group. Immunostaining showed p16 signal in the pyramidal cell of CA1 area in the BCCAO group. CONCLUSION: Vitamin D repairs memory function, senescence expression was lower and NGF was higher in the BCCAO model.

Asunto(s)

Modelos Animales de Enfermedad , ARN Mensajero , Vitamina D , Animales , Ratas , Masculino , Vitamina D/farmacología , ARN Mensajero/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Factor de Crecimiento Nervioso/metabolismo , Factor de Crecimiento Nervioso/genética , Memoria/efectos de los fármacos , Hipocampo/metabolismo , Hipocampo/efectos de los fármacos , Ratas Sprague-Dawley , Factores de Crecimiento Nervioso/metabolismo , Factores de Crecimiento Nervioso/genética , Ataque Isquémico Transitorio/tratamiento farmacológico , Ataque Isquémico Transitorio/metabolismo

RESUMEN

Dry eye disease (DED) is caused by inflammation and damage to the corneal surface due to tear film instability and hyperosmolarity. Various eye drops are used to treat this condition. Each eye drop has different properties and mechanisms of action, so the appropriate drug should be used according to clinical phenotypes. This study aims to compare the therapeutic mechanisms of cyclosporine A (CsA) and diquafosol tetrasodium (DQS). An experimental in vivo/in vitro model of DED using hyperosmolarity showed decreased cell viability, inhibited wound healing, and corneal damage compared to controls. Treatment with cyclosporine or diquafosol restored cell viability and wound healing and reduced corneal damage by hyperosmolarity. The expression of the inflammation-related genes il-1ß, il-1α, and il-6 was reduced by cyclosporine and diquafosol, and the expression of Tnf-α, c1q, and il-17a was reduced by cyclosporine. Increased apoptosis in the DED model was confirmed by increased Bax and decreased Bcl-2 and Bcl-xl expression, but treatment with cyclosporine or diquafosol resulted in decreased apoptosis. Diquafosol increased NGF expression and translocation into the extracellular space. DED has different damage patterns depending on the progression of the lesion. Thus, depending on the type of lesion, eye drops should be selected according to the therapeutic target, focusing on repairing cellular damage when cellular repair is needed or reducing inflammation when inflammation is high and cellular damage is severe.

Asunto(s)

Córnea , Ciclosporina , Modelos Animales de Enfermedad , Síndromes de Ojo Seco , Factor de Crecimiento Nervioso , Nucleótidos de Uracilo , Cicatrización de Heridas , Nucleótidos de Uracilo/farmacología , Factor de Crecimiento Nervioso/metabolismo , Factor de Crecimiento Nervioso/genética , Cicatrización de Heridas/efectos de los fármacos , Animales , Síndromes de Ojo Seco/tratamiento farmacológico , Síndromes de Ojo Seco/metabolismo , Síndromes de Ojo Seco/patología , Córnea/efectos de los fármacos , Córnea/patología , Córnea/metabolismo , Ciclosporina/farmacología , Humanos , Supervivencia Celular/efectos de los fármacos , Apoptosis/efectos de los fármacos , Polifosfatos/farmacología , Ratones

RESUMEN

Peripheral nerve defect are common clinical problem caused by trauma or other diseases, often leading to the loss of sensory and motor function in patients. Autologous nerve transplantation has been the gold standard for repairing peripheral nerve defects, but its clinical application is limited due to insufficient donor tissue. In recent years, the application of tissue engineering methods to synthesize nerve conduits for treating peripheral nerve defect has become a current research focus. This study introduces a novel approach for treating peripheral nerve defects using a tissue-engineered PLCL/SF/NGF@TA-PPy-RGD conduit. The conduit was fabricated by combining electrospun PLCL/SF with an NGF-loaded conductive TA-PPy-RGD gel. The gel, synthesized from RGD-modified tannic acid (TA) and polypyrrole (PPy), provides growth anchor points for nerve cells. In vitro results showed that this hybrid conduit could enhance PC12 cell proliferation, migration, and reduce apoptosis under oxidative stress. Furthermore, the conduit activated the PI3K/AKT signalling pathway in PC12 cells. In a rat model of sciatic nerve defect, the PLCL/SF/NGF@TA-PPy-RGD conduit significantly improved motor function, gastrocnemius muscle function, and myelin sheath axon thickness, comparable to autologous nerve transplantation. It also promoted angiogenesis around the nerve defect. This study suggests that PLCL/SF/NGF@TA-PPy-RGD conduits provide a conducive environment for nerve regeneration, offering a new strategy for peripheral nerve defect treatment, this study provided theoretical basis and new strategies for the research and treatment of peripheral nerve defect.

Asunto(s)

Hidrogeles , Factor de Crecimiento Nervioso , Regeneración Nerviosa , Oligopéptidos , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , Nervio Ciático , Transducción de Señal , Animales , Regeneración Nerviosa/efectos de los fármacos , Ratas , Proteínas Proto-Oncogénicas c-akt/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Transducción de Señal/efectos de los fármacos , Células PC12 , Nervio Ciático/efectos de los fármacos , Nervio Ciático/lesiones , Oligopéptidos/farmacología , Oligopéptidos/química , Hidrogeles/química , Factor de Crecimiento Nervioso/farmacología , Factor de Crecimiento Nervioso/metabolismo , Ratas Sprague-Dawley , Masculino , Proliferación Celular/efectos de los fármacos , Apoptosis/efectos de los fármacos , Andamios del Tejido/química , Ingeniería de Tejidos/métodos , Polímeros/química

RESUMEN

Lower back pain (LBP) is a common condition closely associated with intervertebral disc degeneration (IDD), causing a significant socioeconomic burden. Inflammatory activation in degenerated discs involves pro-inflammatory cytokines, dysregulated regulatory cytokines, and increased levels of nerve growth factor (NGF), leading to further intervertebral disc destruction and pain sensitization. Macrophage polarization is closely related to autophagy. Based on these pathological features, a structured biomimetic nanoparticle coated with TrkA-overexpressing macrophage membranes (TMNP@SR) with a rapamycin-loaded mesoporous silica core is developed. TMNP@SR acted like sponges to adsorbe inflammatory cytokines and NGF and delivers the autophagy regulator rapamycin (RAPA) into macrophages through homologous targeting effects of the outer engineered cell membrane. By regulating autophagy activation, TMNP@SR promoted the M1-to-M2 switch of macrophages to avoid continuous activation of inflammation within the degenerated disc, which prevented the apoptosis of nucleus pulposus cells. In addition, TMNP@SR relieved mechanical and thermal hyperalgesia, reduced calcitonin gene-related peptide (CGRP) and substance P (SP) expression in the dorsal root ganglion, and downregulated GFAP and c-FOS signaling in the spinal cord in the rat IDD model. In summary, TMNP@SR spontaneously inhibits the aggravation of disc inflammation to alleviate disc degeneration and reduce the ingress of sensory nerves, presenting a promising treatment strategy for LBP induced by disc degeneration.

Asunto(s)

Autofagia , Degeneración del Disco Intervertebral , Nanopartículas , Ratas Sprague-Dawley , Degeneración del Disco Intervertebral/tratamiento farmacológico , Degeneración del Disco Intervertebral/metabolismo , Animales , Autofagia/efectos de los fármacos , Nanopartículas/química , Ratas , Masculino , Ratones , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Dolor de la Región Lumbar/tratamiento farmacológico , Materiales Biomiméticos/química , Materiales Biomiméticos/farmacología , Sirolimus/farmacología , Dióxido de Silicio/química , Dióxido de Silicio/farmacología , Núcleo Pulposo/metabolismo , Inflamación/tratamiento farmacológico , Citocinas/metabolismo , Biomimética/métodos , Modelos Animales de Enfermedad , Factor de Crecimiento Nervioso/metabolismo , Células RAW 264.7

RESUMEN

Nerve growth factor (NGF) has long been known as the main ovulation-inducing factor in induced ovulation species, however, recent studies suggested the NGF role also in those with spontaneous ovulation. The first aim of this study was to evaluate the presence and gene expression of NGF and its cognate receptors, high-affinity neurotrophic tyrosine kinase 1 receptor (NTRK1) and low-affinity p75 nerve growth factor receptor (p75NTR), in the ram genital tract. Moreover, the annual trend of NGF seminal plasma values was investigated to evaluate the possible relationship between the NGF production variations and the ram reproductive seasonality. The presence and expression of the NGF/receptors system was evaluated in the testis, epididymis, vas deferens ampullae, seminal vesicles, prostate, and bulbourethral glands through immunohistochemistry and real-time PCR (qPCR), respectively. Genital tract samples were collected from 5 adult rams, regularly slaughtered at a local abattoir. Semen was collected during the whole year weekly, from 5 different adult rams, reared in a breeding facility, with an artificial vagina. NGF seminal plasma values were assessed through the ELISA method. NGF, NTRK1 and p75NTR immunoreactivity was detected in all male organs examined. NGF-positive immunostaining was observed in the spermatozoa of the germinal epithelium, in the epididymis and the cells of the secretory epithelium of annexed glands, NTRK1 receptor showed a localization pattern like that of NGF, whereas p75NTR immunopositivity was localized in the nerve fibers and ganglia. NGF gene transcript was highest (p < 0.01) in the seminal vesicles and lowest (p < 0.01) in the testis than in the other tissues. NTRK1 gene transcript was highest (p < 0.01) in the seminal vesicles and lowest (p < 0.05) in all the other tissues examined. Gene expression of p75NTR was highest (p < 0.01) in the seminal vesicles and lowest (p < 0.01) in the testis and bulbourethral glands. NGF seminal plasma concentration was greater from January to May (p < 0.01) than in the other months. This study highlighted that the NGF system was expressed in the tissues of all the different genital tracts examined, confirming the role of NGF in ram reproduction. Sheep are short-day breeders, with an anestrus that corresponds to the highest seminal plasma NGF levels, thus suggesting the intriguing idea that this factor could participate in an inhibitory mechanism of male reproductive activity, activated during the female anestrus.

Asunto(s)

Genitales Masculinos , Factor de Crecimiento Nervioso , Receptor trkA , Estaciones del Año , Semen , Animales , Masculino , Semen/química , Semen/metabolismo , Receptor trkA/genética , Receptor trkA/metabolismo , Genitales Masculinos/metabolismo , Genitales Masculinos/química , Ovinos/metabolismo , Factor de Crecimiento Nervioso/genética , Factor de Crecimiento Nervioso/metabolismo , Receptor de Factor de Crecimiento Nervioso/genética , Receptor de Factor de Crecimiento Nervioso/metabolismo , Regulación de la Expresión Génica/fisiología

RESUMEN

BACKGROUND AND AIMS: Sodium-glucose cotransporter 2 inhibitors (SGLT2is) can ameliorate arrhythmias; however, the mechanisms underlying their antiarrhythmic effect remain unclear. Therefore, we aimed to test the hypothesis that the SGLT2i empagliflozin (EMPA) ameliorates ventricular arrhythmias caused by myocardial infarction (MI) by inhibiting sympathetic remodeling. METHODS: Male nondiabetic Sprague-Dawley rats were divided into Sham ( n  = 10), MI ( n  = 13), low-EMPA (10 mg/kg/day; n  = 13), and high-EMPA (30 mg/kg/day; n  = 13) groups. Except for the Sham group, MI models were established by ligation of the left anterior descending coronary artery. After 4 weeks, the hearts were removed. Echocardiography, electrical stimulation, hematoxylin-eosin staining and Masson's staining, Western blotting, immunohistochemistry (IHC), and ELISA were performed. RESULTS: Except for left ventricular posterior wall thickness (LVPWT), EMPA treatment significantly ameliorated the left ventricular anterior wall thickness (LVAWT), interventricular septum thickness (IVST), left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), and left ventricular ejection fraction (LVEF) in MI rats; there was no statistical difference between the low-EMPA and high-EMPA groups. The threshold for ventricular fibrillation induction and myocardial fibrosis was significantly ameliorated in EMPA-treated rats, and there was no statistical difference between the high-EMPA and low-EMPA groups. EMPA decreased the expression of nerve growth factor (NGF), tyrosine kinase receptor A (TrkA), tyrosine hydroxylase, and growth-associated protein 43 (GAP43) in the left ventricular infarction margin myocardium of MI rats, especially in the high-EMPA group, with a statistically significant difference between the high-EMPA and low-EMPA groups. High-EMPA significantly decreased noradrenaline (NE) levels in the blood of MI rats; however, there was no statistical difference between the low-EMPA and MI groups. CONCLUSION: EMPA ameliorated the occurrence of ventricular arrhythmias in MI rats, which may be related to a reduction in sympathetic activity, inhibition of the NGF/TrkA pathway, inhibition of sympathetic remodeling, and improvement in cardiac function and cardiac structural remodeling.

Asunto(s)

Compuestos de Bencidrilo , Modelos Animales de Enfermedad , Glucósidos , Infarto del Miocardio , Factor de Crecimiento Nervioso , Ratas Sprague-Dawley , Transducción de Señal , Inhibidores del Cotransportador de Sodio-Glucosa 2 , Sistema Nervioso Simpático , Remodelación Ventricular , Animales , Masculino , Compuestos de Bencidrilo/farmacología , Glucósidos/farmacología , Factor de Crecimiento Nervioso/metabolismo , Infarto del Miocardio/fisiopatología , Infarto del Miocardio/tratamiento farmacológico , Infarto del Miocardio/complicaciones , Infarto del Miocardio/patología , Infarto del Miocardio/metabolismo , Sistema Nervioso Simpático/efectos de los fármacos , Sistema Nervioso Simpático/fisiopatología , Inhibidores del Cotransportador de Sodio-Glucosa 2/farmacología , Remodelación Ventricular/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Receptor trkA/metabolismo , Receptor trkA/antagonistas & inhibidores , Proteína GAP-43/metabolismo , Función Ventricular Izquierda/efectos de los fármacos , Arritmias Cardíacas/etiología , Arritmias Cardíacas/prevención & control , Arritmias Cardíacas/fisiopatología , Arritmias Cardíacas/tratamiento farmacológico , Ratas , Antiarrítmicos/farmacología , Conexina 43

RESUMEN

Chronic pain is a prevalent problem that plagues modern society, and better understanding its mechanisms is critical for developing effective therapeutics. Nerve growth factor (NGF) and its primary receptor, Tropomyosin receptor kinase A (TrkA), are known to be potent mediators of chronic pain, but there is a lack of established methods for precisely perturbing the NGF/TrkA signaling pathway in the study of pain and nociception. Optobiological tools that leverage light-induced protein-protein interactions allow for precise spatial and temporal control of receptor signaling. Previously, our lab reported a blue light-activated version of TrkA generated using light-induced dimerization of the intracellular TrkA domain, opto-iTrkA. In this work, we show that opto-iTrkA activation is able to activate endogenous ERK and Akt signaling pathways and causes the retrograde transduction of phospho-ERK signals in dorsal root ganglion (DRG) neurons. Opto-iTrkA activation also sensitizes the transient receptor potential vanilloid 1 (TRPV1) channel in cellular models, further corroborating the physiological relevance of the optobiological stimulus. Finally, we show that opto-iTrkA enables light-inducible potentiation of mechanical sensitization in mice. Light illumination enables nontraumatic and reversible (<2 days) sensitization of mechanical pain in mice transduced with opto-iTrkA, which provides a platform for dissecting TrkA pathways for nociception in vitro and in vivo.

Asunto(s)

Dolor Crónico , Ganglios Espinales , Luz , Receptor trkA , Animales , Receptor trkA/metabolismo , Dolor Crónico/metabolismo , Ratones , Ganglios Espinales/metabolismo , Canales Catiónicos TRPV/metabolismo , Humanos , Transducción de Señal , Ratones Endogámicos C57BL , Masculino , Factor de Crecimiento Nervioso/metabolismo , Neuronas/metabolismo

RESUMEN

Tropomyosin receptor kinases (Trks) are receptor tyrosine kinases activated by neurotrophic factors, called neurotrophins. Among them, TrkA interacts with the nerve growth factor (NGF), which leads to pain induction. mRNA-display screening was carried out to discover a hit compound 2, which inhibits protein-protein interactions between TrkA and NGF. Subsequent structure optimization improving phosphorylation inhibitory activity and serum stability was pursued using a unique process that took advantage of the peptide being synthesized by translation from mRNA. This gave peptide 19, which showed an analgesic effect in a rat incisional pain model. The peptides described here can serve as a new class of analgesics, and the structure optimization methods reported provide a strategy for discovering new peptide drugs.

Asunto(s)

Receptor trkA , Receptor trkA/antagonistas & inhibidores , Receptor trkA/metabolismo , Animales , Ratas , Humanos , Relación Estructura-Actividad , Descubrimiento de Drogas , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/síntesis química , Analgésicos/farmacología , Analgésicos/química , Analgésicos/síntesis química , Péptidos Cíclicos/farmacología , Péptidos Cíclicos/química , Péptidos Cíclicos/síntesis química , Masculino , Factor de Crecimiento Nervioso/metabolismo , Fosforilación , Dolor/tratamiento farmacológico , Ratas Sprague-Dawley

RESUMEN

Low back pain (LBP) has become a leading cause of disability worldwide. Astrocyte activation in the spinal cord plays an important role in the maintenance of latent sensitization of dorsal horn neurons in LBP. However, the role of spinal c-Jun N-terminal kinase (JNK) in astrocytes in modulating pain behavior of LBP model rats and its neurobiological mechanism have not been elucidated. Here, we investigate the role of the JNK signaling pathway on hypersensitivity and anxiety-like behavior caused by repetitive nerve growth factor (NGF) injections in male non-specific LBP model rats. LBP was produced by two injections (day 0, day 5) of NGF into multifidus muscle of the low backs of rats. We observed prolonged mechanical and thermal hypersensitivity in the low backs or hindpaws. Persistent anxiety-like behavior was observed, together with astrocyte, p-JNK, and neuronal activation and upregulated expression of monocyte chemoattractant protein-1 (MCP-1), and chemokine (C-X-C motif) ligand 1 (CXCL1) proteins in the spinal L2 segment. Second, the JNK inhibitor SP600125 was intrathecally administrated in rats from day 10 to day 12. It attenuated mechanical and thermal hypersensitivity of the low back or hindpaws and anxiety-like behavior. Meanwhile, SP600125 decreased astrocyte and neuronal activation and the expression of MCP-1 and CXCL1 proteins. These results showed that hypersensitivity and anxiety-like behavior induced by NGF in LBP rats could be attenuated by the JNK inhibitor, together with downregulation of spinal astrocyte activation, neuron activation, and inflammatory cytokines. Our results indicate that intervening with the spinal JNK signaling pathway presents an effective therapeutic approach to alleviating LBP.

Asunto(s)

Ansiedad , Dolor de la Región Lumbar , Sistema de Señalización de MAP Quinasas , Ratas Sprague-Dawley , Animales , Masculino , Ratas , Ansiedad/etiología , Dolor de la Región Lumbar/metabolismo , Dolor de la Región Lumbar/etiología , Antracenos/farmacología , Antracenos/uso terapéutico , Hiperalgesia/metabolismo , Astrocitos/metabolismo , Astrocitos/efectos de los fármacos , Factor de Crecimiento Nervioso/metabolismo , Médula Espinal/metabolismo , Quimiocina CCL2/metabolismo , Quimiocina CCL2/genética , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo

RESUMEN

BACKGROUND: Repairation of bone defects remains a major clinical problem. Constructing bone tissue engineering containing growth factors, stem cells, and material scaffolds to repair bone defects has recently become a hot research topic. Nerve growth factor (NGF) can promote osteogenesis of bone marrow mesenchymal stem cells (BMSCs), but the low survival rate of the BMSCs during transplantation remains an unresolved issue. In this study, we investigated the therapeutic effect of BMSCs overexpression of NGF on bone defect by inhibiting pyroptosis. METHODS: The relationship between the low survival rate and pyroptosis of BMSCs overexpressing NGF in localized inflammation of fractures was explored by detecting pyroptosis protein levels. Then, the NGF+/BMSCs-NSA-Sca bone tissue engineering was constructed by seeding BMSCs overexpressing NGF on the allograft bone scaffold and adding the pyroptosis inhibitor necrosulfonamide(NSA). The femoral condylar defect model in the Sprague-Dawley (SD) rat was studied by micro-CT, histological, WB and PCR analyses in vitro and in vivo to evaluate the regenerative effect of bone repair. RESULTS: The pyroptosis that occurs in BMSCs overexpressing NGF is associated with the nerve growth factor receptor (P75NTR) during osteogenic differentiation. Furthermore, NSA can block pyroptosis in BMSCs overexpression NGF. Notably, the analyses using the critical-size femoral condylar defect model indicated that the NGF+/BMSCs-NSA-Sca group inhibited pyroptosis significantly and had higher osteogenesis in defects. CONCLUSION: NGF+/BMSCs-NSA had strong osteogenic properties in repairing bone defects. Moreover, NGF+/BMSCs-NSA-Sca mixture developed in this study opens new horizons for developing novel tissue engineering constructs.

Asunto(s)

Células Madre Mesenquimatosas , Factor de Crecimiento Nervioso , Osteogénesis , Ratas Sprague-Dawley , Andamios del Tejido , Animales , Factor de Crecimiento Nervioso/metabolismo , Factor de Crecimiento Nervioso/genética , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/citología , Ratas , Andamios del Tejido/química , Regeneración Ósea , Aloinjertos , Masculino , Ingeniería de Tejidos/métodos , Piroptosis , Sulfonamidas/farmacología , Diferenciación Celular , Trasplante de Células Madre Mesenquimatosas/métodos , Trasplante Óseo/métodos

RESUMEN

Ovarian theca cells produce testosterone, which acts as a vital precursor substance for synthesizing estrogens during follicular development. Nerve growth factor (NGF) has been shown to participate in reproductive physiology, specifically to follicular development and ovulation. There is currently no available data on the impact of NGF on testosterone synthesis in porcine theca cells. Furthermore, m6A modification is the most common internal modification in eukaryotic mRNAs that are closely associated with female gametogenesis, follicle development, ovulation, and other related processes. It is also uncertain whether the three main enzymes associated with m6A, such as Writers, Erasers, and Readers, play a role in this process. The present study, with an in vitro culture model, investigated the effect of NGF on testosterone synthesis in porcine theca cells and the role of Writers-METTL14 in this process. It was found that NGF activates the PI3K/AKT signaling pathway through METTL14, which regulates testosterone synthesis in porcine theca cells. This study will help to further elucidate the mechanisms by which NGF regulates follicular development and provide new therapeutic targets for ovary-related diseases in female animals. Summary Sentence  The present study investigated the effect of NGF on testosterone synthesis in porcine theca cells. It was found that NGF activates the PI3K/AKT signaling pathway through METTL14, which regulates testosterone synthesis in porcine theca cells.

Asunto(s)

Factor de Crecimiento Nervioso , Testosterona , Células Tecales , Animales , Células Tecales/metabolismo , Células Tecales/efectos de los fármacos , Porcinos , Femenino , Factor de Crecimiento Nervioso/farmacología , Factor de Crecimiento Nervioso/metabolismo , Testosterona/farmacología , Testosterona/biosíntesis , Testosterona/metabolismo , Metiltransferasas/metabolismo , Metiltransferasas/genética , Transducción de Señal/efectos de los fármacos , Células Cultivadas , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo

RESUMEN

A preclinical model of human adolescent binge drinking, adolescent intermittent ethanol exposure (AIE) recreates the heavy binge withdrawal consummatory patterns of adolescents and has identified the loss of basal forebrain cholinergic neurons as a pathological hallmark of this model. Cholinergic neurons of the nucleus basalis magnocellularis (NbM) that innervate the prefrontal cortex (PFC) are particularly vulnerable to alcohol related neurodegeneration. Target derived neurotrophins (nerve growth factor [NGF] and brain-derived neurotrophic factor [BDNF]) regulate cholinergic phenotype expression and survival. Evidence from other disease models implicates the role of immature neurotrophin, or proneurotrophins, activity at neurotrophic receptors in promoting cholinergic degeneration; however, it has yet to be explored in adolescent binge drinking. We sought to characterize the pro- and mature neurotrophin expression, alongside their cognate receptors and cholinergic markers in an AIE model. Male and female Sprague Dawley rats underwent 5 g/kg 20% EtOH or water gavage on two-day-on, two-day-off cycles from post-natal day 25-57. Rats were sacrificed 2 h, 24 h, or 3 weeks following the last gavage, and tissue were collected for protein measurement. Western blot analyses revealed that ethanol intoxication reduced the expression of BDNF and vesicular acetylcholine transporter (vAChT) in the PFC, while NGF was lower in the NbM of AIE treated animals. During acute alcohol withdrawal, proNGF in the PFC was increased while proBDNF decreased, and in the NbM proBDNF increased while NGF decreased. During AIE abstinence, the expression of neurotrophins, their receptors, and vAChT did not differ from controls in the PFC. In contrast, in the NbM the expression of both NGF and choline acetyltransferase (ChAT) were reduced long-term following AIE. Taken together these findings suggest that AIE alters the expression of proneurotrophins and neurotrophins during intoxication and withdrawal that favor prodegenerative mechanisms by increasing the expression of proNGF and proBDNF, while also reducing NGF and BDNF.

Asunto(s)

Núcleo Basal de Meynert , Factor Neurotrófico Derivado del Encéfalo , Etanol , Factor de Crecimiento Nervioso , Corteza Prefrontal , Ratas Sprague-Dawley , Animales , Corteza Prefrontal/metabolismo , Corteza Prefrontal/efectos de los fármacos , Ratas , Masculino , Etanol/farmacología , Femenino , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Factor Neurotrófico Derivado del Encéfalo/biosíntesis , Núcleo Basal de Meynert/efectos de los fármacos , Núcleo Basal de Meynert/metabolismo , Factor de Crecimiento Nervioso/metabolismo , Consumo Excesivo de Bebidas Alcohólicas/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Factores de Crecimiento Nervioso/biosíntesis , Precursores de Proteínas/metabolismo , Precursores de Proteínas/biosíntesis , Neuronas Colinérgicas/efectos de los fármacos , Neuronas Colinérgicas/metabolismo , Consumo de Alcohol en Menores , Modelos Animales de Enfermedad

RESUMEN

One of the most prevalent disorders of the urinary system is urinary tract infection, which is mostly brought on by uropathogenic Escherichia coli (UPEC). The objective of this study was to evaluate the regenerative therapeutic and antibacterial efficacy of PRP for induced bacterial cystitis in dogs in comparison to conventional antibiotics. 25 healthy male mongrel dogs were divided into 5 groups (n = 5). Control negative group that received neither induced infection nor treatments. 20 dogs were randomized into 4 groups after two weeks of induction of UPEC cystitis into; Group 1 (control positive; G1) received weekly intravesicular instillation of sodium chloride 0.9%. Group 2 (syst/PRP; G2), treated with both systemic intramuscular antibiotic and weekly intravesicular instillation of PRP; Group 3 (PRP; G3), treated with weekly intravesicular instillation of PRP, and Group 4 (syst; G4) treated with an intramuscular systemic antibiotic. Animals were subjected to weekly clinical, ultrasonographic evaluation, urinary microbiological analysis, and redox status biomarkers estimation. Urinary matrix metalloproteinases (MMP-2, MMP-9) and urinary gene expression for platelet-derived growth factor -B (PDGF-B), nerve growth factor (NGF), and vascular endothelial growth factor (VEGF) were measured. At the end of the study, dogs were euthanized, and the bladder tissues were examined macroscopically, histologically, and immunohistochemically for NF-κB P65 and Cox-2. The PRP-treated group showed significant improvement for all the clinical, Doppler parameters, and the urinary redox status (p < 0.05). The urinary MMPs activity was significantly decreased in the PRP-treated group and the expression level of urinary NGF and VEGF were downregulated while PDGFB was significantly upregulated (p < 0.05). Meanwhile, the urinary viable cell count was significantly reduced in all treatments (P < 0.05). Gross examination of bladder tissue showed marked improvement for the PRP-treated group, expressed in the histopathological findings. Immunohistochemical analysis revealed a marked increase in Cox-2 and NF-κB P65 in the PRP-treated group (P < 0.05). autologous CaCl2-activated PRP was able to overcome the bacterial infection, generating an inflammatory environment to overcome the old one and initiate tissue healing. Hence, PRP is a promising alternative therapeutic for UPEC cystitis instead of conventional antibiotics.

Asunto(s)

Cistitis , Metaloproteinasa 2 de la Matriz , Metaloproteinasa 9 de la Matriz , Factor de Crecimiento Nervioso , Plasma Rico en Plaquetas , Factor A de Crecimiento Endotelial Vascular , Animales , Perros , Factor de Crecimiento Nervioso/metabolismo , Plasma Rico en Plaquetas/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Cistitis/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/genética , Masculino , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 2 de la Matriz/genética , Modelos Animales de Enfermedad , Escherichia coli Uropatógena/patogenicidad , Infecciones por Escherichia coli , Regulación hacia Abajo , Infecciones Urinarias/tratamiento farmacológico

RESUMEN

Nerve growth factor (NGF) plays a crucial role in cellular growth and neurodifferentiation. To achieve significant neuronal regeneration and repair using in vitro NGF delivery, spatiotemporal control that follows the natural neuronal processes must be developed. Notably, a challenge hindering this is the uncontrolled burst release from the growth factor delivery systems. The rapid depletion of NGF reduces treatment efficacy, leading to poor cellular response. To address this, we developed a highly controllable system using graphene oxygen (GO) and GelMA hydrogels modulated by electrical stimulation. Our system showed superior control over the release kinetics, reducing the burst up 30-fold. We demonstrate that the system is also able to sequester and retain NGF up to 10-times more efficiently than GelMA hydrogels alone. Our controlled release system enabled neurodifferentiation, as revealed by gene expression and immunostaining analysis. The increased retention and reduced burst release from our system show a promising pathway for nerve tissue engineering research toward effective regeneration.

Asunto(s)

Materiales Biocompatibles , Estimulación Eléctrica , Grafito , Hidrogeles , Factor de Crecimiento Nervioso , Regeneración Nerviosa , Hidrogeles/química , Hidrogeles/farmacología , Grafito/química , Grafito/farmacología , Regeneración Nerviosa/efectos de los fármacos , Factor de Crecimiento Nervioso/farmacología , Factor de Crecimiento Nervioso/metabolismo , Factor de Crecimiento Nervioso/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Animales , Tamaño de la Partícula , Ensayo de Materiales , Ratas , Células PC12 , Ingeniería de Tejidos

RESUMEN

Glutamate functions as the major excitatory neurotransmitter for primary sensory neurons and has a crucial role in sensitizing peripheral nociceptor terminals producing sensitization. Glutaminase (GLS) is the synthetic enzyme that converts glutamine to glutamate. GLS-immunoreactivity (-ir) and enzyme activity are elevated in dorsal root ganglion (DRG) neuronal cell bodies during chronic peripheral inflammation, but the mechanism for this GLS elevation is yet to be fully characterized. It has been well established that, after nerve growth factor (NGF) binds to its high-affinity receptor tropomyosin receptor kinase A (TrkA), a retrograde signaling endosome is formed. This endosome contains the late endosomal marker Rab7GTPase and is retrogradely transported via axons to the cell soma located in the DRG. This complex is responsible for regulating the transcription of several critical nociceptive genes. Here, we show that this retrograde NGF signaling mediates the expression of GLS in DRG neurons during the process of peripheral inflammation. We disrupted the normal NGF/TrkA signaling in adjuvant-induced arthritic (AIA) Sprague Dawley rats by the pharmacological inhibition of TrkA or blockade of Rab7GTPase, which significantly attenuated the expression of GLS in DRG cell bodies. The results indicate that NGF/TrkA signaling is crucial for the production of glutamate and has a vital role in the development of neurogenic inflammation. In addition, our pain behavioral data suggest that Rab7GTPase can be a potential target for attenuating peripheral inflammatory pain.

Asunto(s)

Ganglios Espinales , Glutaminasa , Inflamación , Factor de Crecimiento Nervioso , Ratas Sprague-Dawley , Receptor trkA , Transducción de Señal , Animales , Ganglios Espinales/metabolismo , Factor de Crecimiento Nervioso/metabolismo , Glutaminasa/metabolismo , Ratas , Receptor trkA/metabolismo , Inflamación/metabolismo , Inflamación/patología , Masculino , Neuronas/metabolismo , Proteínas de Unión al GTP rab/metabolismo , Proteínas de Unión al GTP rab/genética , Proteínas de Unión a GTP rab7

RESUMEN

Nerve growth factor (NGF), the first neurotrophin to be discovered, has a long and eventful research journey with a series of turning points, setbacks, and achievements. Since the groundbreaking investigations led by Nobel Prize winner Rita Levi-Montalcini, advancements in the comprehension of NGF's functions have revolutionized the field of neuroscience, offering new insights and opportunities for therapeutic innovation. However, the clinical application of NGF has historically been hindered by challenges in determining appropriate dosing, administration strategies, and complications related to the production process. Recent advances in the production and scientific knowledge of recombinant NGF have enabled its clinical development, and in 2018, the United States Food and Drug Administration approved cenegermin-bkbj, a recombinant human NGF, for the treatment of all stages of neurotrophic keratitis. This review traces the evolutionary path that transformed NGF from a biological molecule into a novel therapy with potential research applications beyond the eye. Special emphasis is put on the studies that advanced NGF from discovery to the first medicinal product approved to treat a human disease.

Asunto(s)

Factor de Crecimiento Nervioso , Humanos , Factor de Crecimiento Nervioso/metabolismo , Factor de Crecimiento Nervioso/historia , Animales , Proteínas Recombinantes/uso terapéutico , Proteínas Recombinantes/química , Historia del Siglo XX , Historia del Siglo XXI

RESUMEN

Stress exposure worsens allergic inflammatory diseases substantially. Mast cells (MCs) play a key role in peripheral immune responses to neuroendocrine stress mediators such as nerve growth factor (NGF) and substance P (SP). Mast cell proteases (MCPs) and cholinergic factors (Chrna7, SLURP1) were recently described to modulate MC stress response. We studied MCPs and Chrna7/SLURP1 and their interplay in a mouse model for noise induced stress (NiS) and atopic dermatitis-like allergic inflammation (AlD) and in cultured MC lacking Chrna7. We found that the cholinergic stress axis interacts with neuroendocrine stress mediators and stress-mediator cleaving enzymes in AlD. SP-cleaving mMCP4+ MC were upregulated in AlD and further upregulated by stress in NiS+AlD. Anti-NGF neutralizing antibody treatment blocked the stress-induced upregulation in vivo, and mMCP4+ MCs correlated with measures of AlD disease activity. Finally, high mMCP4 production in response to SP depended on Chrna7/SLURP1 in cultured MCs. In conclusion, mMCP4 and its upstream regulation by Chrna7/SLURP1 are interesting novel targets for the treatment of allergic inflammation and its aggravation by stress.

Asunto(s)

Dermatitis Atópica , Modelos Animales de Enfermedad , Mastocitos , Piel , Receptor Nicotínico de Acetilcolina alfa 7 , Animales , Mastocitos/metabolismo , Mastocitos/inmunología , Dermatitis Atópica/metabolismo , Dermatitis Atópica/patología , Dermatitis Atópica/inmunología , Ratones , Piel/metabolismo , Piel/patología , Receptor Nicotínico de Acetilcolina alfa 7/metabolismo , Inflamación/metabolismo , Inflamación/patología , Péptido Hidrolasas/metabolismo , Activador de Plasminógeno de Tipo Uroquinasa/metabolismo , Sustancia P/metabolismo , Estrés Fisiológico , Ratones Endogámicos C57BL , Factor de Crecimiento Nervioso/metabolismo

RESUMEN

OBJECTIVE: Rotator cuff injury is a common injury that includes inflammation, partial tearing, or complete tearing of the rotator cuff tendon. In cases of rotator cuff tears (RCTs), Tumor Necrosis Factor-alpha (TNF-α) can trigger the release of nerve growth factor (NGF). TNF-α is an important inflammatory mediator that affects rotator cuff activity and increased NGF expression is observed in RCTs. Therefore, this study aimed to investigate whether inhibition of TNF-α could reduce behavioural responses and inflammation levels in rats through NGF. METHODS: A rat RCT model was established, and the CatWalk gait analysis system was used for behavioural assessment. Immunohistochemistry was used to detect NGF protein levels in tendon tissue. Hematoxylin eosin (HE) staining was used to observe histopathological changes. The expressions of Interleukin-1beta (IL-1ß) and Cyclooxygenase-2 (COX2) were detected by western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR). The expression of apoptosis protein Bcl-2-associated X (Bax), B-cell lymphoma 2 (Bcl-2), and Cysteine-aspartic acid protease-3 (Caspase-3) were detected using WB. Oxidative stress markers, namely Reactive Oxygen Species (ROS), Malondialdehyde (MDA), and Superoxide Dismutase (SOD) were quantified in tissues using an ELISA kit. RESULTS: In the RCT model, elevated NGF protein expression, noticeable atrophy in the supraspinatus muscle tissue, and substantial fat infiltration were observed. The levels of IL-1ß, COX2, apoptosis, and oxidative stress were all increased. TNF-α inhibition resulted in decreased NFG expression, decreased tissue fibrosis, and improved tendon atrophy. Moreover, when TNF-α was inhibited, the expressions of IL-1ß and Cox2 were reduced and both apoptosis and oxidative stress were decreased. The results showed that inhibiting TNF-α had the potential to reduce inflammation levels and behavioural responses in rats. CONCLUSION: TNF-α can affect behaviour and inflammation in rats with RCTs through NGF, and TNF-α inhibition can improve rotator cuff injury.

Asunto(s)

Inflamación , Factor de Crecimiento Nervioso , Ratas Sprague-Dawley , Lesiones del Manguito de los Rotadores , Factor de Necrosis Tumoral alfa , Animales , Factor de Crecimiento Nervioso/metabolismo , Lesiones del Manguito de los Rotadores/patología , Lesiones del Manguito de los Rotadores/metabolismo , Ratas , Factor de Necrosis Tumoral alfa/metabolismo , Inflamación/metabolismo , Inflamación/patología , Masculino , Estrés Oxidativo/efectos de los fármacos , Conducta Animal/efectos de los fármacos , Modelos Animales de Enfermedad , Apoptosis/efectos de los fármacos , Interleucina-1beta/metabolismo , Ciclooxigenasa 2/metabolismo

RESUMEN

Spinal pain affects individuals of all ages and is the most common musculoskeletal problem globally. Its clinical management remains a challenge as the underlying mechanisms leading to it are still unclear. Here, we report that significantly increased numbers of senescent osteoclasts (SnOCs) are observed in mouse models of spinal hypersensitivity, like lumbar spine instability (LSI) or aging, compared to controls. The larger population of SnOCs is associated with induced sensory nerve innervation, as well as the growth of H-type vessels, in the porous endplate. We show that deletion of senescent cells by administration of the senolytic drug Navitoclax (ABT263) results in significantly less spinal hypersensitivity, spinal degeneration, porosity of the endplate, sensory nerve innervation, and H-type vessel growth in the endplate. We also show that there is significantly increased SnOC-mediated secretion of Netrin-1 and NGF, two well-established sensory nerve growth factors, compared to non-senescent OCs. These findings suggest that pharmacological elimination of SnOCs may be a potent therapy to treat spinal pain.

Asunto(s)

Senescencia Celular , Osteoclastos , Animales , Ratones , Osteoclastos/metabolismo , Osteoclastos/efectos de los fármacos , Osteoclastos/fisiología , Senescencia Celular/efectos de los fármacos , Células Receptoras Sensoriales/efectos de los fármacos , Células Receptoras Sensoriales/fisiología , Células Receptoras Sensoriales/metabolismo , Modelos Animales de Enfermedad , Masculino , Factor de Crecimiento Nervioso/metabolismo , Factor de Crecimiento Nervioso/farmacología , Netrina-1/metabolismo , Netrina-1/genética , Ratones Endogámicos C57BL