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Wound healing is a complex process involving cell proliferation, migration, and extracellular matrix (ECM) remodeling. Extracellular vesicles (EVs) or exosomes derived from adipose tissue-derived stem cells (ASCs) are emerging as promising alternatives to cell therapy for advanced wound healing. Hyaluronic acid (HA), a major component of the skin ECM, is widely utilized in wound dressings and dermal fillers. This study aimed to investigate the effects of ASC-derived exosomes (ASC-EXOs) on human dermal fibroblasts (HDFs) and their potential combination with HA in in vivo wound healing and dermal filler models. In HDFs, ASC-EXOs increased cell proliferation and migration. ASC-EXOs also upregulated the expression of genes involved in cell proliferation and wound healing while stimulating collagen production in HDFs. In a porcine wound healing model, topical treatment with a combination of HA and ASC-EXOs led to higher wound closure rates compared to HA alone. Histological examination showed increased re-epithelialization and collagen type III deposition in wounds treated with the combination of HA and ASC-EXOs. In a mouse dermal filler model, tissues injected with the combination of HA and ASC-EXOs exhibited thicker tissue layers, increased vascularization, enhanced infiltration of myofibroblasts, and higher levels of collagen III and collagen fiber content compared to HA alone. These findings suggest that ASC-EXOs have beneficial effects on cell proliferation, migration, and gene expression related to wound healing, and they may accelerate wound closure and promote tissue regeneration. Furthermore, the combination of HA and ASC-EXOs may enhance wound healing and tissue remodeling, indicating its potential for both clinical and regenerative aesthetic applications in skin repair and regeneration.
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Preenchedores Dérmicos , Exossomos , Células-Tronco Mesenquimais , Camundongos , Humanos , Animais , Suínos , Exossomos/metabolismo , Preenchedores Dérmicos/metabolismo , Cicatrização/genética , Células-Tronco Mesenquimais/metabolismo , Tecido Adiposo , Colágeno/metabolismoRESUMO
Free fatty acids receptor 3 (FFA3, GPR41) and 2 (FFA2, GPR43), for which the short-chain fatty acids (SCFAs) acetate and propionate are agonist, have emerged as important G-protein-coupled receptors influenced by diet and gut flora composition. A recent study (Kimura et al., 2011) demonstrated functional expression of FFA3 in the rodent sympathetic nervous system (SNS) providing a potential link between nutritional status and autonomic function. However, little is known of the source of endogenous ligands, signaling pathways, or effectors in sympathetic neurons. In this study, we found that FFA3 and FFA2 are unevenly expressed in the rat SNS with higher transcript levels in prevertebral (e.g., celiac-superior mesenteric and major pelvic) versus paravertebral (e.g., superior cervical and stellate) ganglia. FFA3, whether heterologously or natively expressed, coupled via PTX-sensitive G-proteins to produce voltage-dependent inhibition of N-type Ca(2+) channels (Cav2.2) in sympathetic neurons. In addition to acetate and propionate, we show that ß-hydroxybutyrate (BHB), a metabolite produced during ketogenic conditions, is also an FFA3 agonist. This contrasts with previous interpretations of BHB as an antagonist at FFA3. Together, these results indicate that endogenous BHB levels, especially when elevated under certain conditions, such as starvation, diabetic ketoacidosis, and ketogenic diets, play a potentially important role in regulating the activity of the SNS through FFA3.
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Ácido 3-Hidroxibutírico/farmacologia , Canais de Cálcio Tipo N/efeitos dos fármacos , Neurônios/fisiologia , Receptores Acoplados a Proteínas G/agonistas , Sistema Nervoso Simpático/fisiologia , Animais , DNA Complementar/biossíntese , DNA Complementar/genética , Fenômenos Eletrofisiológicos/fisiologia , Transferência Ressonante de Energia de Fluorescência , Gânglios Simpáticos/citologia , Gânglios Simpáticos/efeitos dos fármacos , Gânglios Simpáticos/fisiologia , Células HeLa , Humanos , Hibridização In Situ , Corpos Cetônicos/farmacologia , Ligantes , Masculino , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Sistema Nervoso Simpático/citologia , TransfecçãoRESUMO
BACKGROUND: Androgenetic alopecia (AGA) is a prevalent hair loss disorder with psychological repercussions. Traditional treatments have limitations, leading to the exploration of regenerative therapies such as exosomes derived from adipose tissue stem cells (ASC-Exosomes). METHODS: First, using human hair follicle (HF) dermal papilla cells (hDPCs) treated with ASC-Exosomes, ALP, VCAN, ß-catenin, and LEF-1 levels with RT-PCR and p-GSK3ß, GSK3ß, ß-catenin, ALP, and ß-actin levels with western blot analysis were assessed. Hair shaft elongation test and assay for ALP, Ki-67, and ß-catenin were done using human HF organ culture. Patients with AGA had ASC-Exosomes treatment and were evaluated for hair counts, photographic assessments, subjective satisfaction, and safety profiles. RESULTS: ASC-Exosomes impact hDPCs, increasing proliferation and the upregulation of hair growth-related genes, including ALP, VCAN, ß-catenin, and LEF-1. The Wnt/ß-catenin pathway was activated, indicating their role in promoting hair growth. ASC-Exosomes also promoted hair shaft elongation and ALP activity, suggesting a potential for hair regeneration. Thirty participants with AGA enrolled and treated over 24 weeks. The subjects experienced a significant increase in total hair density, improved global photographic assessments, and reported higher subjective satisfaction without severe adverse reactions. CONCLUSION: This research contributes to the growing body of evidence supporting the use of exosomes in hair loss treatment, offering a safe and effective alternative for individuals with AGA.
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Recently, particulate matter (PM) has been shown to exacerbate atopic dermatitis (AD) by inducing an inflammatory response. Meanwhile, several studies revealed that exosomes derived from adipose tissue-derived mesenchymal stem cells promote wound healing and alleviate inflammation via their regenerative and immunomodulatory capacities. Our study aimed to investigate the effects of human adipose tissue-derived mesenchymal stem cell-derived (ASC)-exosomes in PM-induced AD. An AD-like triple-cell model was established by treating human keratinocytes, dermal fibroblasts, and mast cells with polyinosinic:polycytidylic acid (Poly I:C) and interleukin 1 alpha (IL-1α). The effects of PM and ASC-exosomes on the expression of pro-inflammatory cytokines and skin barrier proteins were examined using quantitative real-time polymerase chain reaction, western blotting, and immunofluorescence. PM increased pro-inflammatory cytokines (IL-6, IL-1ß, and IL-1α) and decreased the anti-inflammatory cytokine IL-10, while the mRNA expression of skin barrier proteins (loricrin and filaggrin) decreased. However, when the cells were treated with ASC-exosomes, the PM-induced effects on pro-inflammatory cytokines and skin barrier proteins were reversed. Our results confirmed that PM-induced inflammation and skin barrier damage were alleviated by ASC-exosomes in our AD-like triple-cell model. These data suggest that ASC-exosomes can serve as a therapeutic agent for PM-exacerbated AD.
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Dermatite Atópica , Exossomos , Humanos , Dermatite Atópica/terapia , Dermatite Atópica/tratamento farmacológico , Exossomos/metabolismo , Material Particulado/toxicidade , Material Particulado/metabolismo , Citocinas/metabolismo , Inflamação/metabolismo , Tecido Adiposo/metabolismo , Pele/metabolismoRESUMO
Electrically excitable cells have voltage-dependent ion channels on the plasma membrane that regulate membrane permeability to specific ions. Voltage-gated Ca(2+) channels (VGCCs) are especially important as Ca(2+) serves as both a charge carrier and second messenger. Zebrafish (Danio rerio) are an important model vertebrate for studies of neuronal excitability, circuits, and behavior. However, electrophysiological properties of zebrafish VGCCs remain largely unexplored because a suitable preparation for whole cell voltage-clamp studies is lacking. Rohon-Beard (R-B) sensory neurons represent an attractive candidate for this purpose because of their relatively large somata and functional homology to mammalian dorsal root ganglia (DRG) neurons. Transgenic zebrafish expressing green fluorescent protein in R-B neurons, (Isl2b:EGFP)(ZC7), were used to identify dissociated neurons suitable for whole cell patch-clamp experiments. Based on biophysical and pharmacological properties, zebrafish R-B neurons express both high- and low-voltage-gated Ca(2+) current (HVA- and LVA-I(Ca), respectively). Ni(+)-sensitive LVA-I(Ca) occur in the minority of R-B neurons (30%) and ω-conotoxin GVIA-sensitive Ca(V)2.2 (N-type) Ca(2+) channels underlie the vast majority (90%) of HVA-I(Ca). To identify G protein coupled receptors (GPCRs) that modulate HVA-I(Ca), a panel of neurotransmitters was screened. Application of GABA/baclofen or serotonin produced a voltage-dependent inhibition while application of the mu-opioid agonist DAMGO resulted in a voltage-independent inhibition. Unlike in mammalian neurons, GPCR-mediated voltage-dependent modulation of I(Ca) appears to be transduced primarily via a cholera toxin-sensitive Gα subunit. These results provide the basis for using the zebrafish model system to understanding Ca(2+) channel function, and in turn, how Ca(2+) channels contribute to mechanosensory function.
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Canais de Cálcio/fisiologia , Células Receptoras Sensoriais/citologia , Células Receptoras Sensoriais/fisiologia , Peixe-Zebra/fisiologia , Animais , Animais Geneticamente Modificados , Baclofeno/farmacologia , Canais de Cálcio/efeitos dos fármacos , Ala(2)-MePhe(4)-Gly(5)-Encefalina/farmacologia , Proteínas de Fluorescência Verde/genética , Modelos Animais , Neurotransmissores/farmacologia , Técnicas de Patch-Clamp , Ácido gama-Aminobutírico/farmacologiaRESUMO
Endocannabinoids (eCB) such as 2-arachidonylglycerol (2-AG) are lipid metabolites that are synthesized in a postsynaptic neurons and act upon CB(1) cannabinoid receptors (CB(1)R) in presynaptic nerve terminals. This retrograde transmission underlies several forms of short and long term synaptic plasticity within the CNS. Here, we constructed a model system based on isolated rat sympathetic neurons, in which an eCB signaling cascade could be studied in a reduced, spatially compact, and genetically malleable system. We constructed a complete eCB production/mobilization pathway by sequential addition of molecular components. Heterologous expression of four components was required for eCB production and detection: metabotropic glutamate receptor 5a (mGluR5a), Homer 2b, diacylglycerol lipase alpha, and CB(1)R. In these neurons, application of l-glutamate produced voltage-dependent modulation of N-type Ca(2+) channels mediated by activation of CB(1)R. Using both molecular dissection and pharmacological agents, we provide evidence that activation of mGluR5a results in rapid enzymatic production of 2-AG followed by activation of CB(1)R. These experiments define the critical elements required to recapitulate retrograde eCB production and signaling in a single peripheral neuron. Moreover, production/mobilization of eCB can be detected on a physiologically relevant time scale using electrophysiological techniques. The system provides a platform for testing candidate molecules underlying facilitation of eCB transport across the plasma membrane.
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Ácidos Araquidônicos/metabolismo , Glicerídeos/metabolismo , Modelos Neurológicos , Neurônios/metabolismo , Receptores de Glutamato Metabotrópico/metabolismo , Gânglio Cervical Superior/metabolismo , Animais , Cálcio/metabolismo , Proteínas de Transporte/metabolismo , Endocanabinoides , Ácido Glutâmico/metabolismo , Proteínas de Arcabouço Homer , Humanos , Lipase Lipoproteica/genética , Lipase Lipoproteica/metabolismo , Masculino , Técnicas de Patch-Clamp , Nervos Periféricos/metabolismo , Ratos , Ratos Wistar , Receptor CB1 de Canabinoide/metabolismo , Receptor de Glutamato Metabotrópico 5 , Transdução de Sinais , Fatores de TempoRESUMO
OBJECTIVE: To compare the characteristics and outcomes of patients who underwent surgical treatment for stage 2 medication-related osteonecrosis of the jaw (MRONJ) versus osteomyelitis. METHODS: This retrospective study compared the following variables in 73 patients with stage 2 MRONJ versus 89 patients with osteomyelitis: impaired wound healing after surgery, sex, age, the presence of actinomycosis, location of the jaw lesion, and involvement of the inferior alveolar nerve (IAN). RESULTS: There were significant differences between the groups in age, sex, rates of impaired wound healing, actinomycosis, and the location (anterior/posterior) of the lesion. Impaired wound healing after surgical treatment in the stage 2 MRONJ group was associated with patient age. All patients with impaired wound healing after the initial surgery recovered fully after reoperation. CONCLUSIONS: These findings for surgical treatment of stage 2 MRONJ may help clinicians plan surgical treatment of MRONJ.
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Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/cirurgia , Osteomielite/cirurgia , Actinomicose , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/fisiopatologia , Feminino , Humanos , Masculino , Nervo Mandibular , Pessoa de Meia-Idade , Osteomielite/fisiopatologia , Reoperação , Estudos Retrospectivos , Índice de Gravidade de Doença , Resultado do Tratamento , CicatrizaçãoRESUMO
Efforts to apply augmented reality (AR) technology in the medical field include the introduction of AR techniques into dental practice. The present report introduces a simple method of applying AR during an inferior alveolar nerve block, a procedure commonly performed in dental clinics.
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The present case report describes a complication involving facial blanching symptoms occurring during inferior alveolar nerve block anesthesia (IANBA). Facial blanching after IANBA can be caused by the injection of an anesthetic into the maxillary artery area, affecting the infraorbital artery.
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Infection that progresses to deep areas of the neck requires appropriate assessment of the airway, and securing of the airway is critical in patients with deep neck infection. In the patient in our case report, bilateral pneumothorax occurred while performing tracheostomy to the airways of a patient with deep neck infection, and therefore, this paper details the method used to secure the airway of patients with deep neck infection.
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PURPOSE: To analyze the relative position of the mandibular foramina (MnFs) in patients diagnosed with skeletal class III malocclusion. MATERIALS AND METHODS: Computed tomography (CT) images were collected from 85 patients. The vertical lengths of each anatomic point from the five horizontal planes passing through the MnF were measured at the coronoid process, sigmoid notch, condyle, and the gonion. The distance from the anterior ramus point to the posterior ramus point on the five horizontal planes was designated the anteroposterior horizontal distance of the ramus for each plane. The perpendicular distance from each anterior ramus point to each vertical plane through the MnF was designated the horizontal distance from the anterior ramus to the MnF. The horizontal and vertical positions were examined by regression analysis. RESULTS: Regression analysis showed the heights of the coronoid process, sigmoid notch, and condyle for the five horizontal planes were significantly related to the height of the MnF, with the highest significance associated with the MnF-mandibular plane (coefficients of determination (R(2)): 0.424, 0.597, and 0.604, respectively). The horizontal anteroposterior length of the ramus and the distance from the anterior ramus point to the MnF were significant by regression analysis. CONCLUSION: The relative position of the MnF was significantly related to the vertical heights of the sigmoid notch, coronoid process, and condyle as well as to the horizontal anteroposterior length of the ascending ramus. These findings should be clinically useful for patients with skeletal class III mandibular prognathism.
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Obesity is closely associated with metabolic disorders such as hyperglycemia and dyslipidemia. Leptin-deficient ob/ob mice (C57BL/6J-ob/ob) and C57BL/6J mice were randomly assigned to a diet of black rice with giant embryo (BR), white rice (WR), or AIN-93G (control) and pair-fed for 14 weeks. Although there was no significant difference in body weight, BR-fed ob/ob mice had (1) significantly lower body fat mass than WR- and control-fed ob/ob mice determined by dual-energy X-ray absorptiometry; (2) significantly lower blood glucose, serum insulin, and triacylglycerol levels than control-fed ob/ob mice; and (3) significantly lower liver weight, hepatic triacylglycerol, and hepatic lipid droplets than both WR- and control-fed ob/ob mice. Furthermore, DNA damage in the liver, determined by phosphorylated H2AX protein, and in the kidney, determined by single-cell gel electrophoresis, was significantly lower in BR-fed than WR- and control-fed ob/ob mice. This study indicates that BR ameliorates obesity and its related metabolic disorders.
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Obesidade/metabolismo , Oryza/química , Animais , Dieta , Ácidos Graxos/metabolismo , Feminino , Leptina/metabolismo , Fígado/metabolismo , Masculino , Doenças Metabólicas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Triglicerídeos/metabolismoRESUMO
The Cav1.2 Ca2+ channel is essential for cardiac and smooth muscle contractility and many physiological functions. We mutated single, double, and quadruple sites of the four potential Asn (N)-glycosylation sites in the rabbit Cav1.2 into Gln (Q) to explore the effects of Nglycosylation. When a single mutant (N124Q, N299Q, N1359Q, or N1410Q) or Cav1.2/WT was expressed in Xenopus oocytes, the biophysical properties of single mutants were not significantly different from Cav1.2/WT. In comparison, the double mutant N124,299Q showed a positive shift in voltage-dependent gating. Furthermore, the quadruple mutant (QM; N124,299,1359,1410Q) showed a positive shift in voltage-dependent gating as well as a reduction of current. We tagged EGFP to the QM, double mutants, and Cav1.2/WT to chase the mechanisms underlying the reduced currents of QM. The surface fluorescence intensity of QM was weaker than that of Cav1.2/WT, suggesting that the reduced current of QM arises from its lower surface expression than Cav1.2/WT. Tunicamycin treatment of oocytes expressing Cav1.2/WT mimicked the effects of the quadruple mutations. These findings suggest that Nglycosylation contributes to the surface expression and voltage-dependent gating of Cav1.2.
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Canais de Cálcio Tipo L/metabolismo , Processamento de Proteína Pós-Traducional , Animais , Fenômenos Biofísicos , Canais de Cálcio Tipo L/genética , Análise Mutacional de DNA , Glicosilação , Mutação de Sentido Incorreto , Mutação Puntual , Coelhos , XenopusRESUMO
FFAR3 (GPR41) is a G-protein coupled receptor for which short-chain fatty acids serve as endogenous ligands. The receptor is found on gut enteroendocrine L-cells, pancreatic ß-cells, and sympathetic neurons, and is implicated in obesity, diabetes, allergic airway disease, and altered immune function. In primates, FFAR3 is segmentally duplicated resulting in GPR42, a gene currently classified as a suspected pseudogene. In this study, we sequenced FFAR3 and GPR42 open reading frames from 56 individuals and found an unexpectedly high frequency of polymorphisms contributing to several complex haplotypes. We also identified a frequent (18.8%) structural variation that results in GPR42 copy number polymorphism. Finally, sequencing revealed that 50.6% of GPR42 haplotypes differed from FFAR3 by only a single non-synonymous substitution and that the GPR42 reference sequence matched only 4.4% of the alleles. Sequencing of cDNA from human sympathetic ganglia and colon revealed processed transcripts matching the GPR42 genotype. Expression of several GPR42 haplotypes in rat sympathetic neurons revealed diverse pharmacological phenotypes that differed in potency and efficacy. Our data suggest that GPR42 be reclassified as a functioning gene and that recognition of sequence and copy number polymorphism of the FFAR3/GPR42 complex be considered during genetic and pharmacological investigation of these receptors.
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Variações do Número de Cópias de DNA/genética , Pseudogenes , Receptores Acoplados a Proteínas G/genética , Animais , Gânglios Simpáticos/efeitos dos fármacos , Gânglios Simpáticos/metabolismo , Genótipo , Haplótipos/genética , Humanos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Ratos , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/biossínteseRESUMO
Distortion of intracellular oxidant and antioxidant balances appears to be a common feature that underlies in age-related male sexual impairment. Therefore regulating oxidative defense mechanisms might be an ideal approach in improving male sexual dysfunctions. In the present study, the effect of Korean red ginseng aqueous extract (KRG) on age-induced testicular dysfunction in rats was investigated. KRG (200mg/kg) mixed with regular pellet diet was administered orally for six months and the morphological, spermatogenic and antioxidant enzyme status in testis of aged rats (18months) were evaluated. Data indicated a significant change in morphology and decrease in spermatogenesis-related parameters in aged rats (AC) compared with young rats (YC). Sperm number, germ cell count, Sertoli cell count and Sertoli cell index were significantly (p<0.05) restored in KRG-treated aged rat groups (G-AC). Further the increased lipid peroxidation as measured by malondialdehyde (p<0.05), and altered enzymatic (superoxide dismutase, glutathione peroxidase, glutathione S-transferase, glutathione reductase and catalase) and non-enzymatic (reduced glutathione, ascorbic acid and α-tocopherol) antioxidants (p<0.05) were attenuated by KRG treatment in aged rats to near normal levels as in YC groups. Furthermore, proteomic analysis demonstrated differential expression of selected proteins such as phosphatidylinositol transfer protein, fatty acid binding protein-9, triosephosphate isomerase-1 and aldehyde (aldose) reductase-1in aged rats was significantly (p<0.05) protected by KRG treatment. In conclusion, long-term administration of KRG restored aging-induced testicular ineffectiveness in rats by modulating redox proteins and oxidative defense mechanisms.
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Peroxidação de Lipídeos/efeitos dos fármacos , Oxirredução/efeitos dos fármacos , Panax , Extratos Vegetais/farmacologia , Maturação do Esperma/efeitos dos fármacos , Envelhecimento/fisiologia , Animais , Antioxidantes/farmacologia , Modelos Animais de Doenças , Disfunção Erétil/tratamento farmacológico , Glutationa Peroxidase/metabolismo , Masculino , Ratos , Espermatogênese/efeitos dos fármacos , Espermatozoides/metabolismo , Superóxido Dismutase/metabolismo , Resultado do TratamentoRESUMO
IL-4 is emerging as a candidate cytokine for the treatment of inflammatory and autoimmune diseases. We have reported that IL-4 has anti-angiogenic activity and inhibits the growth of human umbilical vein endothelial cells (HUVEC) in response to vascular endothelial growth factor (VEGF) or fibroblast growth factor-2 (FGF-2). Cell cycle analysis of this effect revealed that IL-4 arrests the growth of FGF-2-stimulated HUVEC in G0 + G1 phases. The absence of subdiploid cells showed that it did not induce apoptosis. Growth arrest was dose-dependent, but the percentage of G0 + G1 phase cells never exceeded 85%. An immunoblot analysis demonstrated that expression of p53 and p21(Waf1) was increased and that of cyclin D1 and cyclin E decreased by IL-4. These results show that IL-4 inhibits endothelial cell growth by altering the expression of cell cycle regulatory molecules.
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Ciclo Celular/fisiologia , Ciclina D1/metabolismo , Ciclina E/metabolismo , Ciclinas/metabolismo , Células Endoteliais/metabolismo , Interleucina-4/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Células Cultivadas , Inibidor de Quinase Dependente de Ciclina p21 , Células Endoteliais/citologia , Fator 2 de Crescimento de Fibroblastos/metabolismo , Humanos , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
The goal of this study was to elucidate the involvement of neuronal and glial calcium-binding proteins in the stimulation of gamma-aminobutyric acid (GABA) transport system by kainate-induced excitotoxicity in the rat retina. We used immunohistochemical method to assess the localization of GABA reuptake and calcium-binding proteins. After systemic administration of kainate, the neuronal GABA transporter does not show an association with calbindin D-28K. However, the localization of the GAT-3 transport system in Müller glial cells is closely correlated with the S100 proteins interacting with glial fibrillary acidic protein (GFAP) in response to kainate injury. Furthermore, we demonstrate that kainate-mediated excitotoxicity induced concomitant distribution shift of glial GABA transporter, S100 proteins and GFAP in the distal processes and endfeet of glial cells during the first 48 h.
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Proteínas de Transporte/metabolismo , Agonistas de Aminoácidos Excitatórios/toxicidade , Ácido Caínico/toxicidade , Proteínas de Membrana/metabolismo , Neuroglia/metabolismo , Transportadores de Ânions Orgânicos , Retina/efeitos dos fármacos , Proteínas S100/metabolismo , Animais , Calbindinas , Proteínas da Membrana Plasmática de Transporte de GABA , Proteína Glial Fibrilar Ácida/metabolismo , Imuno-Histoquímica , Proteínas de Membrana Transportadoras/metabolismo , Neuroglia/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Retina/lesões , Retina/metabolismo , Proteína G de Ligação ao Cálcio S100/metabolismo , Fatores de TempoRESUMO
Although nerve injury is known to up- and down-regulate some metabotropic receptors in vagal afferent neurons of the nodose ganglia (NG), the functional significance has not been elucidated. In the present study, thus, we examined whether nerve injury affected receptor-mediated Ca2+ channel modulation in the NG neurons. In this regard, unilateral vagotomy was performed using male Sprague-Dawley rats. One week after vagotomy, Ca2+ currents were recorded using the whole-cell variant of patch-clamp technique in enzymatically dissociated NG neurons. In sham controls, norepinephrine (NE)-induced Ca2+ current inhibition was negligible. Following vagotomy, however, the NE responses were dramatically increased. This phenomenon was in accordance with up-regulation of alpha2A/B-adrenergic receptor mRNAs as quantified using real-time RT-PCR analysis. In addition, neuropeptide Y (NPY) and prostaglandin E2 responses were moderately augmented in vagotomized NG neurons. The altered NPY response appears to be caused by up-regulation of Y2 receptors negatively coupled to Ca2+ channels. In contrast, nerve injury significantly suppressed opioid (tested with DAMGO)-induced Ca2+ current inhibition with down-regulation of micro-receptors. Taken together, these results demonstrated for the first time that the profile of neurotransmitter-induced Ca2+ channel modulation is significantly altered in the NG neurons under pathophysiological state of nerve injury.
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Canais de Cálcio/metabolismo , Neurônios Aferentes/fisiologia , Gânglio Nodoso/fisiopatologia , Animais , Células Cultivadas , Dinoprostona/metabolismo , Masculino , Potenciais da Membrana/fisiologia , Neurônios Aferentes/patologia , Neuropeptídeo Y/metabolismo , Gânglio Nodoso/lesões , Técnicas de Patch-Clamp , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Receptores Adrenérgicos alfa 2/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , VagotomiaRESUMO
The root of Panax ginseng improves testicular function both in humans and animals. However, the molecular mechanism by which ginseng exerts this effect has not been elucidated. Changes in protein expression in the rat testis in response to a pectinase-treated P. ginseng extract (GINST) were identified using 2-dimensional electrophoresis (2-DE) and MALDI-TOF/TOF MS. Number of sperm, Sertoli cells and germ cells, and the Sertoli Cell Index decrease in the testis of aged rats (AR) relative to young control rats (YCR). However, those parameters were completely restored in GINST-treated AR (GINST-AR). A proteomic analysis identified 14 proteins that were differentially expressed between vehicle-treated AR (V-AR) and GINST-AR. Out of these, the expression of glutathione-S-transferase (GST) mu5 and phospholipid hydroperoxide (PH) glutathione peroxidase (GPx) was significantly up-regulated in GINST-AR compared to V-AR. The activity of GPx and GST, as well as the expression of glutathione, in the testis of GINST-AR was higher than that in V-AR. The levels of lipid peroxidation (LPO) increased in AR compared with YCR, but this change was reversed by GINST-AR. These results suggest that the administration of GINST enhances testicular function by elevating GPx and GST activity, thus resulting in increased glutathione, which prevents LPO in the testis.
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Envelhecimento/fisiologia , Panax , Fitoterapia/métodos , Testículo/efeitos dos fármacos , Envelhecimento/efeitos dos fármacos , Envelhecimento/patologia , Animais , Antioxidantes/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Glutationa Peroxidase/metabolismo , Glutationa Transferase/metabolismo , Hormônios Esteroides Gonadais/sangue , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Oxirredução , Extratos Vegetais/farmacologia , Poligalacturonase , Proteoma/metabolismo , Ratos , Ratos Sprague-Dawley , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Espermatogênese/fisiologia , Testículo/metabolismo , Testículo/patologia , Testículo/fisiopatologiaRESUMO
RGK proteins, Gem, Rad, Rem1, and Rem2, are members of the Ras superfamily of small GTP-binding proteins that interact with Ca2+ channel ß subunits to modify voltage-gated Ca2+ channel function. In addition, RGK proteins affect several cellular processes such as cytoskeletal rearrangement, neuronal dendritic complexity, and synapse formation. To probe the phylogenetic origins of RGK protein-Ca2+ channel interactions, we identified potential RGK-like protein homologs in genomes for genetically diverse organisms from both the deuterostome and protostome animal superphyla. RGK-like protein homologs cloned from Danio rerio (zebrafish) and Drosophila melanogaster (fruit flies) expressed in mammalian sympathetic neurons decreased Ca2+ current density as reported for expression of mammalian RGK proteins. Sequence alignments from evolutionarily diverse organisms spanning the protostome/deuterostome divide revealed conservation of residues within the RGK G-domain involved in RGK protein--Cavß subunit interaction. In addition, the C-terminal eleven residues were highly conserved and constituted a signature sequence unique to RGK proteins but of unknown function. Taken together, these data suggest that RGK proteins, and the ability to modify Ca2+ channel function, arose from an ancestor predating the protostomes split from deuterostomes approximately 550 million years ago.