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1.
Nature ; 577(7790): 392-398, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31915380

RESUMO

More than twelve morphologically and physiologically distinct subtypes of primary somatosensory neuron report salient features of our internal and external environments1-4. It is unclear how specialized gene expression programs emerge during development to endow these subtypes with their unique properties. To assess the developmental progression of transcriptional maturation of each subtype of principal somatosensory neuron, we generated a transcriptomic atlas of cells traversing the primary somatosensory neuron lineage in mice. Here we show that somatosensory neurogenesis gives rise to neurons in a transcriptionally unspecialized state, characterized by co-expression of transcription factors that become restricted to select subtypes as development proceeds. Single-cell transcriptomic analyses of sensory neurons from mutant mice lacking transcription factors suggest that these broad-to-restricted transcription factors coordinate subtype-specific gene expression programs in subtypes in which their expression is maintained. We also show that neuronal targets are involved in this process; disruption of the prototypic target-derived neurotrophic factor NGF leads to aberrant subtype-restricted patterns of transcription factor expression. Our findings support a model in which cues that emanate from intermediate and final target fields promote neuronal diversification in part by transitioning cells from a transcriptionally unspecialized state to transcriptionally distinct subtypes by modulating the selection of subtype-restricted transcription factors.


Assuntos
Neurogênese , Neurônios/fisiologia , Animais , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Camundongos , Fator de Crescimento Neural/metabolismo , Neurônios/citologia , RNA/análise , RNA/genética , Análise de Célula Única , Fator de Transcrição Brn-3B/genética , Fator de Transcrição Brn-3B/metabolismo , Fator de Transcrição Brn-3C/genética , Fator de Transcrição Brn-3C/metabolismo
2.
J Photochem Photobiol B ; 204: 111785, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31954267

RESUMO

Postoperative sensory disturbances of inferior alveolar nerve (IAN) are major challenges in dental procedures. We aimed to investigate the effect of photobiomodulation therapy (PBMT) with 810 nm and 980 nm diode lasers on behavioral and immunological factors in a rat IAN crush model. Seventy-two rats were randomly assigned to the four groups of 810 nm laser (crush injury+810 nm laser; 6 J/cm2, 15 sessions, every 48 h), 980 nm laser (crush injury+980 nm laser; same protocol), control (crush injury without irradiation), and sham surgery (no crush injury and no irradiation). The neurosensory response of IAN was evaluated by Von Frey behavioral test before (baseline) and post-surgery in a period of one month. Changes of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), nuclear factor-kappa B (NF-κB), TNF-α, and IL-1ß, were assessed on days 2 and 30 post injury. Data were analyzed for significant differences by repeated measures and one-way ANOVA (p < .05). One day after surgery, all rats subjected to nerve injury showed significant increase in the withdrawal threshold of von Frey test compared to the baseline (p = .02 for control and p = .03 for laser groups). The threshold gradually returned to the baseline scores in 810 nm, 980 nm, and control groups from days 11, 17, and 29, respectively. There was a significant lower withdrawal threshold in 810 nm and 980 nm laser groups compared to the control group in days 11 to 19 and 9 to 23, respectively. At both time points, the levels of NGF and BDNF were significantly higher in 810 nm laser group compared to the control group. There was a significant difference between laser and control groups regarding NF-κB expression (all p values<.001). TNF-α and IL-1ß were significantly lower in laser groups compared to the control group (all p values < .001). PBMT with 810 and 980 nm diode laser protocol used in this study, promoted the neurosensory recovery of IAN after crush injury in rats. In addition, application of 810 nm diode laser was associated with more improvement in immunological responses compared to that of 980 nm laser.


Assuntos
Lasers Semicondutores , Nervo Mandibular/efeitos da radiação , Animais , Comportamento Animal/efeitos da radiação , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Modelos Animais de Doenças , Regulação para Baixo/efeitos da radiação , Interleucina-1beta/metabolismo , Terapia com Luz de Baixa Intensidade , Masculino , Nervo Mandibular/imunologia , Nervo Mandibular/metabolismo , Traumatismos do Nervo Mandibular/imunologia , Traumatismos do Nervo Mandibular/metabolismo , Traumatismos do Nervo Mandibular/radioterapia , NF-kappa B/metabolismo , Fator de Crescimento Neural/metabolismo , Ratos , Ratos Wistar , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima/efeitos da radiação
3.
Am J Physiol Cell Physiol ; 318(2): C360-C371, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31774700

RESUMO

Nerve growth factor (NGF) is recognized as a pleiotropic molecule, exerting a variety of biological effects on different cell types and pathophysiological conditions, and its role in tissue wound healing has been recently highlighted. However, the preferential cellular target of NGF is still elusive in the complex cellular and molecular cross talk that accompanies wound healing. Thus, to explore possible NGF cellular targets in skin wound healing, we investigated the in vitro NGF responsiveness of keratinocytes (cell line HEKa), fibroblasts (cell line BJ), and endothelial cells (cell line HUVEC), also in the presence of adverse microenvironmental conditions, e.g., hyperglycemia. The main results are summarized as follows: 1) NGF stimulates keratinocyte proliferation and HUVEC proliferation and angiogenesis in a dose-dependent manner although it has no effect on fibroblast proliferation; 2) NGF stimulates keratinocyte but not fibroblast migration in the wound healing assay; and 3) NGF completely reverts the proliferation impairment of keratinocytes and the angiogenesis impairment of HUVECs induced by high d-glucose concentration in the culture medium. These results contribute to better understanding possible targets for the therapeutic use of NGF in skin repair.


Assuntos
Células Endoteliais/metabolismo , Fibroblastos/metabolismo , Queratinócitos/metabolismo , Fator de Crescimento Neural/metabolismo , Cicatrização/fisiologia , Animais , Linhagem Celular , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Pele/metabolismo
4.
Dis Markers ; 2019: 7510315, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31827642

RESUMO

NGF and VEGF are known to be involved in different psychiatric diseases. In order to verify hints from basic research that both neurotrophines interact with each other, serum levels of NGF and VEGF were measured in a cohort of 33 healthy individuals and correlated. NGF level was 126.30 pg/mL (±155.43), and VEGF level was 57.28 pg/mL (±44.48). Both factors were significantly correlated, confirming their interaction and legitimising the usage of their respective ratio (0.8 (±0.42)) as a less varying additional marker in prospective studies.


Assuntos
Fator de Crescimento Neural/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto , Estudos de Coortes , Feminino , Voluntários Saudáveis , Humanos , Masculino , Domínios e Motivos de Interação entre Proteínas
5.
BMC Gastroenterol ; 19(1): 221, 2019 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-31856738

RESUMO

BACKGROUND: Nerve growth factor (NGF) and enteric glial cells (EGCs) are associated with visceral hypersensitivity and gastrointestinal motility disorder, which may represent the pathogenesis of functional dyspepsia (FD). This study aimed to investigate the expression of NGF, its high affinity receptor tropomyosin receptor kinase A (TrkA) and the EGC activation marker glial fibrillary acidic protein (GFAP) in the gastric mucosa of patients with FD and the association of these proteins with dyspeptic symptoms. METHODS: Gastric mucosal biopsies taken from 27 FD patients (9 epigastric pain syndrome (EPS) patients, 7 postprandial distress syndrome (PDS) patients and 11 EPS overlap PDS patients) and 26 control subjects were used for analysis. The expression of NGF, TrkA and GFAP was examined, and the association of these proteins with dyspeptic symptoms, including epigastric pain, postprandial fullness, early satiation and epigastric burning, was analysed. RESULTS: The expression levels of NGF, TrkA, and GFAP in the gastric mucosa were significantly higher in the EPS group, the PDS group, and the EPS overlap PDS group than in the healthy control group. There was no significant difference between the FD subgroups. TrkA colocalized with GFAP, which indicated that TrkA was localized to EGCs, and the expression of TrkA in EGCs was significantly higher in the FD group than in the control group. Changes in the expression of NGF, TrkA, and GFAP were positively correlated with epigastric pain, postprandial fullness and early satiation but had no significant relationship with epigastric burning. CONCLUSIONS: The increased expression of gastric NGF, TrkA and GFAP might be involved in FD pathophysiology and symptom perception.


Assuntos
Dispepsia/metabolismo , Mucosa Gástrica/metabolismo , Proteína Glial Fibrilar Ácida/metabolismo , Fator de Crescimento Neural/metabolismo , Receptor trkA/metabolismo , Dor Abdominal/metabolismo , Adulto , Estudos de Casos e Controles , Dispepsia/etiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
6.
Zhongguo Zhen Jiu ; 39(11): 1205-10, 2019 Nov 12.
Artigo em Chinês | MEDLINE | ID: mdl-31724358

RESUMO

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expressions of growth arrest-specific protein 7 (Gas7) and nerve growth factor (NGF) in arcuate nucleus (ARC) of rats with focal cerebral ischemia and explore the potential action mechanism of EA in treatment of focal cerebral ischemia. METHODS: A total of 50 SD rats were randomized into 4 groups, named a normal group (n =12), a sham-operation group (n =12), a model group (n =14) and an EA group (n =12). In the model group and the EA group, the thread embolization method was adopted to duplicate the model of the right middle cerebral arterial embolism. In the sham-operation group, the skin of the neck was opened and sutured without any other intervention. In the EA group, EA was applied to "Baihui" (CV 20) and "Zusanli" (ST 36) on the left side, once a day, 30 min each time, consecutively for 21 days, while there was no any intervention in the normal group, the sham-operation group and the model group. Using the immunohistochemistry (IHC) method and Western blot method, the expressions of Gas7 and NFG of ARC on the ischemic side were determined. Using Nissle staining, the morphological changes in ARC neurons were observed. RESULTS: The results of Nissle staining showed that there was no significant change in the morphology of ARC neurons in the normal group and the sham-operation group. In the model group, the volume of neuron cells was atrophied obviously and the cells were arranged irregularly. In the EA group, the morphology of ARC neuron was similar to the normal group. The results of IHC and Western blot indicated that the expressions of immunoreactive neurons and protein of Gas7 and NGF in ARC of the rats in the model group were increased obviously as compared with the normal group and the sham-operation group and the expressions in the EA group were further enhanced as compared with the model group (all P<0.05). CONCLUSION: Gas7 and NGF may be participated in the compensatory process of partial protection of the body in the patients with focal cerebral ischemia. EA up-regulates the expressions of Gas7 and NGF in ARC, which may be one of the neuroprotective mechanisms of EA in treatment of cerebral ischemia.


Assuntos
Isquemia Encefálica , Infarto Cerebral , Eletroacupuntura , Fator de Crescimento Neural/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Animais , Isquemia Encefálica/metabolismo , Isquemia Encefálica/terapia , Infarto Cerebral/metabolismo , Infarto Cerebral/terapia , Humanos , Ratos , Ratos Sprague-Dawley
7.
Int J Mol Sci ; 20(23)2019 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-31775361

RESUMO

Metastases in thyroid cancer are associated with aggressive disease and increased patient morbidity, but the factors driving metastatic progression are unclear. The precursor for nerve growth factor (proNGF) is increased in primary thyroid cancers, but its expression or significance in metastases is not known. In this study, we analysed the expression of proNGF in a retrospective cohort of thyroid cancer lymph node metastases (n = 56), linked with corresponding primary tumours, by automated immunohistochemistry and digital quantification. Potential associations of proNGF immunostaining with clinical and pathological parameters were investigated. ProNGF staining intensity (defined by the median h-score) was significantly higher in lymph node metastases (h-score 94, interquartile range (IQR) 50-147) than in corresponding primary tumours (57, IQR 42-84) (p = 0.002). There was a correlation between proNGF expression in primary tumours and corresponding metastases, where there was a 0.68 (95% CI 0 to 1.2) increase in metastatic tumour h-score for each unit increase in the primary tumour h-score. However, larger tumours (both primary and metastatic) had lower proNGF expression. In a multivariate model, proNGF expression in nodal metastases was negatively correlated with lateral neck disease and being male. In conclusion, ProNGF is expressed in locoregional metastases of thyroid cancer and is higher in lymph node metastases than in primary tumours, but is not associated with high-risk clinical features.


Assuntos
Adenocarcinoma Folicular/secundário , Biomarcadores Tumorais/metabolismo , Carcinoma Papilar/secundário , Fator de Crescimento Neural/metabolismo , Precursores de Proteínas/metabolismo , Neoplasias da Glândula Tireoide/patologia , Adenocarcinoma Folicular/metabolismo , Adulto , Idoso , Carcinoma Papilar/metabolismo , Feminino , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Estudos Retrospectivos , Neoplasias da Glândula Tireoide/metabolismo
8.
BMC Complement Altern Med ; 19(1): 295, 2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31694615

RESUMO

BACKGROUND: NGF-TrkA is well known to play a key role in propagating and sustaining pruritogenic signals, which form the pathology of chronic pruritus. Inhibition of NGF-TrkA is a known strategy for the treatment of pruritus. In the present paper, we describe the identification, in vitro characterization, structure-activity analysis, and inhibitory evaluation of a novel TrkA inhibitory scaffold exemplified by Cucurbitacins (Cus). METHODS: Cus were identified as TrkA inhibitors in a large-scale kinase library screen. To obtain structural models of Cus as TrkA inhibitors, AutoDock was used to explore their binding to TrkA. Furthermore, PC12 cell culture systems have been used to study the effects of Cus and traditional Chinese medicinal plants (Tian Gua Di and bitter gourd leaf) extracts on the kinase activity of TrkA. RESULTS: Cus block the phosphorylation of TrkA on several tyrosine sites, including Tyr490, Tyr674/675, and Tyr785, and inhibit downstream Akt and MAPK phosphorylation in response to NGF in PC12 cell model systems. Furthermore, traditional Chinese medicinal plants (Tian Gua Di and bitter gourd leaf) containing Cu extracts were shown to inhibit the phosphorylation of TrkA and Akt. These data reveal mechanisms, at least partly, of the anti-pruritus bioactivity of Cus. CONCLUSION: Taken together, with the recent discovery of the important role of TrkA as a therapeutic target, Cus could be the basis for the design of improved TrkA kinase inhibitors, which could someday help treat pruritus.


Assuntos
Cucumis melo/química , Cucurbitacinas/química , Inibidores Enzimáticos/química , Momordica charantia/química , Extratos Vegetais/química , Receptor trkA/antagonistas & inibidores , Motivos de Aminoácidos , Animais , Frutas/química , Humanos , Cinética , Fator de Crescimento Neural/metabolismo , Células PC12 , Fosforilação , Ratos , Receptor trkA/química
9.
Reprod Biol Endocrinol ; 17(1): 93, 2019 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-31718673

RESUMO

BACKGROUND: The nerve growth factor (NGF), a member of the neurotrophins family, plays an important role not only in the nervous but also in other non-nervous systems such as the reproductive system. The aim of the paper is to study the in vitro effect of NGF on rabbit sperm functions. METHODS: Ten adult rabbit bucks were collected five times, and pooled semen samples have been analysed. NGF was quantified in seminal plasma, and the distribution of NGF receptors (TrKA and p75NTR) in sperm was established. Moreover, the dose-effect of NGF on motility rate and track speed was evaluated. Successively, the effect of the neutralisation of NGF receptors was assessed to verify the specific role of each receptor. Untreated sperm were used as control. RESULTS: Our study identified several interesting results: i) We detected NGF in seminal plasma and TrKA and p75NTR in sperm surface. In particular, TrKA is localised in the head and p75NTR in the midpiece and tail of rabbit sperm. ii) Once the optimal dose of NGF (100 ng/mL) was established, its addition affected both kinetics and other physiological traits (capacitation, apoptosis and necrosis) of rabbit sperm. (iii) The neutralisation of TrKA and p75NTR receptors affected sperm traits differently. In particular, sperm speed, apoptosis and capacitation seemed mainly modulated via p75NTR receptor, whereas motile, live cells, necrosis and acrosome reaction were modulated via TrKA. CONCLUSION: For the first time, we showed the presence of p75NTR in rabbit sperm. NGF affects kinetic and other physiological traits of rabbit sperm. Most of these changes are modulated by the receptors involved (TrKA or p75NTR). Considering that some seminal disorders in human have been correlated with a lower NGF concentration and no studies have been done on the possible involvement of NGF receptors, these findings also provide new insights on human fertility.


Assuntos
Apoptose/efeitos dos fármacos , Fator de Crescimento Neural/farmacologia , Sêmen/metabolismo , Capacitação Espermática/efeitos dos fármacos , Motilidade Espermática/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Animais , Humanos , Masculino , Fator de Crescimento Neural/metabolismo , Coelhos , Receptor trkA/metabolismo , Receptores de Fator de Crescimento Neural/metabolismo , Análise do Sêmen/métodos , Cabeça do Espermatozoide/metabolismo , Cauda do Espermatozoide/metabolismo , Espermatozoides/fisiologia
10.
FEBS Open Bio ; 9(12): 2063-2071, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31605506

RESUMO

As one of the main neurotrophic factors, nerve growth factor (NGF) participates in various processes related to viability, plasticity, and neuronal growth. NGF is known to protect against cell death and toxicity triggered by ß-amyloid (Aß), but the underlying mechanism remains unclear. Here, we investigated this process in SKNSH neuroblastoma, in which NGF reduced cell death induced by Aß25-35. Furthermore, NGF suppressed the production of reactive oxygen species (ROS) and promoted antioxidant function via Aß25-35. Additionally, we demonstrated that NGF impaired the activation of the JNK/c-Jun signaling pathway and significantly increased Nrf2 nuclear translocation and HO-1 expression. Nrf2 elimination abolished the protective effect of NGF-1 on Aß25-35-induced ROS generation, apoptosis, and activation of the JNK/c-Jun pathway. The results of our study indicate that NGF protects neuroblastoma against injury triggered by Aß25-35 via suppression of ROS-JNK/c-Jun pathway stimulation through the Nrf2/HO-1 pathway.


Assuntos
Apoptose/fisiologia , Fator de Crescimento Neural/metabolismo , Neuroblastoma/metabolismo , Peptídeos beta-Amiloides/metabolismo , Animais , Antioxidantes , Morte Celular , Linhagem Celular Tumoral , Sobrevivência Celular , Heme Oxigenase-1/metabolismo , Heme Oxigenase-1/fisiologia , Humanos , Camundongos , Fator 2 Relacionado a NF-E2/metabolismo , Fator 2 Relacionado a NF-E2/fisiologia , Fator de Crescimento Neural/fisiologia , Fármacos Neuroprotetores , Estresse Oxidativo , Fragmentos de Peptídeos/metabolismo , Cultura Primária de Células , Espécies Reativas de Oxigênio , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia
11.
PLoS One ; 14(9): e0222602, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31557169

RESUMO

Pentosan polysulphate sodium (PPS) is a promising therapeutic agent for blocking knee pain in individuals with knee osteoarthritis (KOA). The mode of action of PPS in this context is unknown. We hypothesised that the osteocyte, being the principal cell type in the sub-chondral bone, was capable of expressing the pain mediator Nerve Growth Factor (NGF), and that this may be altered in the presence of PPS. We tested the expression of NGF and the response to PPS in the presence or absence of the proinflammatory cytokine tumour necrosis factor-alpha (TNFα), in human osteocytes. For this we differentiated human primary osteoblasts grown from subchondral bone obtained at primary knee arthroplasty for KOA to an osteocyte-like stage over 28d. We also tested NGF expression in fresh osteocytes obtained by sequential digestion from KOA bone and by immunofluorescence in KOA bone sections. We demonstrate for the first time the production and secretion of NGF/proNGF by this cell type derived from patients with KOA, implicating osteocytes in the pain response in this pathological condition and possibly others. PPS inhibited TNFα-induced levels of proNGF secretion and TNFα induced NGF mRNA expression. Together, this provides evidence that PPS may act to suppress the release of NGF in the subchondral bone to ameliorate pain associated with knee osteoarthritis.


Assuntos
Artralgia/tratamento farmacológico , Fator de Crescimento Neural/metabolismo , Osteoartrite do Joelho/tratamento farmacológico , Osteócitos/metabolismo , Poliéster Sulfúrico de Pentosana/farmacologia , Idoso , Artralgia/etiologia , Feminino , Humanos , Osteoartrite do Joelho/complicações , Osteócitos/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia
12.
Life Sci ; 235: 116844, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31499069

RESUMO

AIMS: 10-O-(N,N-dimethylaminoethyl)-ginkgolide B methanesulfonate (XQ-1H), a new derivative of ginkgolide B, has drawn great attention for its potent bioactivities against ischemia-induced injury. The purpose of this study was to further investigate the effect of XQ-1H against acute ischemic stroke by inducing middle cerebral artery occlusion/reperfusion (MCAO/R) injuries in mice. MAIN METHODS: Treatment of XQ-1H (78 or 39 mg/kg, i.g., bid) 2 h after MCAO improved motor skills and ameliorated the severity of brain infarction and apoptosis seen in the mice by diminishing pathological changes and the activation of a pro-apoptotic protein Cleaved-Caspase-3, which in turn induced anti-apoptotic Bcl-xL. Through introducing Wnt/ß-catenin signaling inhibitor XAV-939, XQ-1H was proven to intensively promoted neurogenesis in the peri-infarct cortex, subventricular area (SVZ) and the dentate gyrus (DG) subgranular area (SGZ) in a Wnt signal dependent way by compromising the activation of GSK3ß, which in turn upregulated Wnt1, ß-catenin, Neuro D1 and Cyclin D1, most possibly through the activation of PI3K/Akt signaling via the upregulation of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF). KEY FINDINGS: We conclude that XQ-1H preserved the motor functions, limited apoptosis, and concomitantly promoted neurogenesis-related protein expression by Wnt signaling-dependently compromising GSK3ß/Caspase-3 activity and enhancing the expression of Wnt1/ß-catenin/Neuro D1/Cyclin D1 and Bcl-xL. SIGNIFICANCE: This research may benefit the development of stroke therapeutics targeting neurogenesis through Wnt upregulation by XQ-1H.


Assuntos
Apoptose/efeitos dos fármacos , Isquemia Encefálica/tratamento farmacológico , Ginkgolídeos/farmacologia , Ginkgolídeos/uso terapêutico , Lactonas/farmacologia , Lactonas/uso terapêutico , Neurogênese/efeitos dos fármacos , Via de Sinalização Wnt/efeitos dos fármacos , beta Catenina/metabolismo , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Encéfalo/patologia , Isquemia Encefálica/complicações , Isquemia Encefálica/patologia , Caspase 3/metabolismo , Ciclina D1/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Compostos Heterocíclicos com 3 Anéis/farmacologia , Infarto da Artéria Cerebral Média , Masculino , Camundongos , Fator de Crescimento Neural/metabolismo , Recuperação de Função Fisiológica/efeitos dos fármacos , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/tratamento farmacológico , Acidente Vascular Cerebral/patologia , Regulação para Cima/efeitos dos fármacos , Proteína bcl-X/metabolismo
13.
J Pharmacol Sci ; 140(3): 255-262, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31402210

RESUMO

Heat shock protein 90 (HSP90) antagonists are currently being evaluated as potential anticancer drugs. However, adverse effects related to these drugs, such as fatigue and pain, suggest that they affect neurons. Therefore, to understand the influence of HSP90 inhibitors on neurons, we investigated the effects of geldanamycin, an HSP90 antagonist, on nerve growth factor (NGF)-differentiated pheochromocytoma 12 (PC12) cells, particularly, on the expression and phosphorylation of proteins and kinases in the NGF pathway. Geldanamycin significantly inhibited NGF-induced neurite outgrowth and phosphorylation of Akt and extracellular signal-related kinase 1/2 in PC12 cells. Furthermore, geldanamycin inhibited the phosphorylation of collapsin response mediator protein 2 and the expression of cyclin-dependent kinase 5 in the presence of NGF, but did not significantly affect the expression of glycogen synthase kinase 3ß. These results suggest that geldanamycin influences microtubule-binding proteins and kinases relating to neurite outgrowth, thereby inducing neuronal impairment.


Assuntos
Neoplasias das Glândulas Suprarrenais/tratamento farmacológico , Benzoquinonas/farmacologia , Diferenciação Celular/efeitos dos fármacos , Lactamas Macrocíclicas/farmacologia , Fator de Crescimento Neural/metabolismo , Neuritos/efeitos dos fármacos , Crescimento Neuronal/efeitos dos fármacos , Feocromocitoma/tratamento farmacológico , Neoplasias das Glândulas Suprarrenais/metabolismo , Animais , Linhagem Celular Tumoral , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neuritos/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Células PC12 , Feocromocitoma/metabolismo , Fosforilação/efeitos dos fármacos , Ratos , Transdução de Sinais/efeitos dos fármacos
14.
J Orthop Res ; 37(11): 2389-2400, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31286562

RESUMO

Intervertebral disc (IVD) degeneration is a major contributor to chronic low back pain and is characterized by decreases in cellularity and proteoglycan synthesis, upregulation of matrix degradation, and increases in pro-inflammatory factors with neurovascular invasion. Current treatments fail to target the underlying pathology or promote tissue repair and approaches such as viral transfection raise safety concerns due to mutagenesis and unwarranted immune responses. To avoid such concerns, nonviral transfection is a viable method of gene delivery into the host cell while bypassing the caveats of viral delivery. Brachyury is expressed in the developing notochord and is associated with an immature healthy nucleus pulposus (NP). We hypothesize that Brachyury can reprogram degenerate NP cells to a healthy pro-anabolic phenotype with increased proteoglycan content and decreased expression of catabolic, inflammatory, and neurovascular markers. NP cells obtained from human autopsy and surgical tissues were transfected with plasmids encoding for Brachyury or an empty vector control via bulk electroporation. Post transfection, cells were seeded in three-dimensional agarose constructs cultured over 4 weeks and analyzed for viability, gene expression, and proteoglycan. Results demonstrated successful transfection of both autopsy and surgical NP cells. We observed long-term Brachyury expression, significant increased expression of NP phenotypic markers FOXF1, KRT19, and chondrogenic marker SOX9 with decreases in inflammatory cytokines IL1-ß/IL6, NGF, and MMPs and significant increases in glycosaminoglycan accumulation. These results highlight nonviral transfection with developmental transcription factors, such as Brachyury, as a promising method to reprogram degenerate human disc cells toward a healthy NP phenotype. Clinical significance: This project proposes a novel translational approach for the treatment of intervertebral disc degeneration via direct reprogramming of diseased human patient-derived IVD cells to a healthy phenotype. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2389-2400, 2019.


Assuntos
Proteínas Fetais/genética , Terapia Genética/métodos , Degeneração do Disco Intervertebral/terapia , Núcleo Pulposo/metabolismo , Proteínas com Domínio T/genética , Transfecção/métodos , Adulto , Idoso , Citocinas/metabolismo , Feminino , Proteínas Fetais/metabolismo , Expressão Gênica , Glicosaminoglicanos/metabolismo , Humanos , Masculino , Metaloproteinases da Matriz/metabolismo , Pessoa de Meia-Idade , Fator de Crescimento Neural/metabolismo , Cultura Primária de Células , Estudo de Prova de Conceito , Proteínas com Domínio T/metabolismo , Adulto Jovem
15.
Nat Commun ; 10(1): 3307, 2019 07 24.
Artigo em Inglês | MEDLINE | ID: mdl-31341171

RESUMO

Current therapies for most neurodegenerative disorders are only symptomatic in nature and do not change the course of the disease. Gene therapy plays an important role in disease modifying therapeutic strategies. Herein, we have designed and optimized a series of highly branched poly(ß-amino ester)s (HPAEs) containing biodegradable disulfide units in the HPAE backbone (HPAESS) and guanidine moieties (HPAESG) at the extremities. The optimized polymers are used to deliver minicircle DNA to multipotent adipose derived stem cells (ADSCs) and astrocytes, and high transfection efficiency is achieved (77% in human ADSCs and 52% in primary astrocytes) whilst preserving over 90% cell viability. Furthermore, the top-performing candidate mediates high levels of nerve growth factor (NGF) secretion from astrocytes, causing neurite outgrowth from a model neuron cell line. This synergistic gene delivery system provides a viable method for highly efficient non-viral transfection of ADSCs and astrocytes.


Assuntos
Doenças Neurodegenerativas/genética , Transfecção/métodos , Astrócitos/metabolismo , Terapia Genética/métodos , Humanos , Células-Tronco Mesenquimais , Fator de Crescimento Neural/metabolismo , Doenças Neurodegenerativas/terapia , Polímeros/química
16.
Life Sci ; 233: 116698, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31356906

RESUMO

AIM: Type 1 diabetes (T1DM) is a common chronic disease in childhood. Increasing insulin resistance in puberty gives rise to higher doses of insulin usage in treatment. Of this reason new approaches in treatment are needed. Noopept researches suggest it to have anti-diabetic properties. We tried to determine the effects of noopept on pubertal diabetes. MAIN METHOD: The research was made with 60 prepubertal, 28 day-old, male, Sprague Dawley rats. The rats were divided into randomised 6 groups (n = 10/group). i) Control, ii) Diabetes Control, iii) Noopept Control, iv) Diabetes + Noopept, v) Diabetes + Insulin, vi) Diabetes + Insulin + Noopept. T1DM model was induced by streptozotocin on postnatal 28th day. 0.5 mg/kg noopept and 1 IU insulin were administered intraperitoneally for 14 days. Blood glucose and body weight measurements, puberty follow-up and MWM tests were performed. Hippocampus, hypothalamus and testis were evaluated histologically. Hypothalamic GnRH and kisspeptin were studied immunohistochemically. Serum LH, FSH and insulin, hippocampal homogenate NGF and BDNF levels were determined by ELISA. KEY FINDINGS: Delayed puberty was normalized by noopept (p < 0.05). Blood glucose levels were lower in noopept-administered diabetic groups (p < 0.05). Noopept decreased HOMA-IR in insulin administered diabetic group (p < 0.05). Number of degenerated cells in hippocampus and testis were higher in diabetes control group when compared with other groups (p < 0.05). GnRH immunoreactivity in Diabetes + Noopept group was increased when compared to insulin + noopept group (p = 0.018). There was no difference in kisspeptin, serum LH, FSH, hippocampal NGF-BDNF levels and spatial learning assessment among groups (p > 0.05). SIGNIFICANCE: Noopept may have positive effect in treatment of pubertal diabetes.


Assuntos
Cognição/fisiologia , Diabetes Mellitus Experimental/tratamento farmacológico , Dipeptídeos/farmacologia , Resistência à Insulina , Fármacos Neuroprotetores/farmacologia , Puberdade/fisiologia , Animais , Glicemia/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Cognição/efeitos dos fármacos , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Insulina/metabolismo , Masculino , Fator de Crescimento Neural/metabolismo , Puberdade/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
17.
Neurosci Lett ; 707: 134308, 2019 08 10.
Artigo em Inglês | MEDLINE | ID: mdl-31153972

RESUMO

Acori Tatarinowii Rhizoma (ATR, the dried rhizome of Acorus tatarinowii Schott.) is a traditional Chinese medicine widely used to treat brain diseases, e.g. depression, forgetfulness, anxiety and epilepsy. Several lines of evidence support that ATR has neuronal beneficial functions in animal models, but its action mechanism in cellular level is unknown. Here, we identified α-asarone and ß-asarone could be the major active ingredients of ATR, which, when applied onto cultured rat astrocytes, significantly stimulated the expression and secretion of neurotrophic factors, i.e. nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and glial derived neurotrophic factor (GDNF), in dose-dependent manners. These results suggested that the neuronal action of ATR, triggered by asarone, might be mediated by an increase of expression of neurotrophic factors in astrocytes, which therefore could support the clinical usage of ATR. In addition, application of PKA inhibitor, H89, in cultured astrocytes partially blocked the asarone-induced neurotrophic factor expression, suggesting the involvement of PKA signaling. The results proposed that α-asarone and ß-asarone from ATR could serve as potential candidates for drug development in neurodegenerative diseases.


Assuntos
Acorus/química , Anisóis/farmacologia , Astrócitos/efeitos dos fármacos , Medicamentos de Ervas Chinesas/química , Fatores de Crescimento Neural/metabolismo , Animais , Anisóis/isolamento & purificação , Astrócitos/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Células Cultivadas , Expressão Gênica , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Fator de Crescimento Neural/metabolismo , Ratos Sprague-Dawley , Rizoma/química
18.
Biomed Res Int ; 2019: 6539294, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31240219

RESUMO

Chemotherapy leads to a loss of fertility and reproductive endocrine function, thereby increasing the risk of premature ovarian failure (POF). Studies have suggested that the transplantation of mesenchymal stem cells could inhibit apoptosis in ovarian granulosa cells and improve follicular development. In the present study, the effects of human umbilical cord mesenchymal stem cell (UCMSC) transplantation on ovarian function after ovarian damage caused by chemotherapy and the mechanism underlying these effects were investigated. POF model rats were obtained by the intraperitoneal injection of cyclophosphamide, and cultured UCMSCs were transplanted by tail vein injection. Serum estrogen, follicle-stimulating hormone, gonadotropin releasing hormone, and anti-Mullerian hormone levels were detected by ELISA. Folliculogenesis was evaluated by histopathological examination. The expression levels of nerve growth factor (NGF), high affinity nerve growth factor receptor (TrkA), follicle-stimulating hormone receptor (FSHR), and caspase-3 were evaluated by western blotting and RT-qPCR. The natural reproductive capacity was assessed by pregnant rate and numbers of embryos. The results indicated that UCMSC transplantation recovered disturbed hormone secretion and folliculogenesis in POF rats. NGF and TrkA levels increased, while FSHR and caspase-3 decreased. The pregnancy rate of POF rats was improved. Therefore, UCMSCs could reduce ovarian failure due to premature senescence caused by chemotherapy, and the NGF/TrkA signaling pathway was involved in the amelioration of POF.


Assuntos
Antineoplásicos/efeitos adversos , Transplante de Células-Tronco Mesenquimais/métodos , Fator de Crescimento Neural/metabolismo , Insuficiência Ovariana Primária/terapia , Cordão Umbilical/transplante , Animais , Hormônio Antimülleriano/sangue , Apoptose/efeitos dos fármacos , Caspase 3/sangue , Ciclofosfamida/efeitos adversos , Estrogênios/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Liberador de Gonadotropina/sangue , Humanos , Células-Tronco Mesenquimais , Fator de Crescimento Neural/sangue , Ovário/patologia , Gravidez , Insuficiência Ovariana Primária/induzido quimicamente , Ratos , Ratos Sprague-Dawley , Receptores do FSH/sangue
19.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(5): 385-392, 2019 May.
Artigo em Chinês | MEDLINE | ID: mdl-31223106

RESUMO

Objective To investigate the effect of low-level laser on the polarization and secretory phenotype of primary cultured M1 bone marrow-derived macrophages (BMDMs) in neuronal axons of dorsal root ganglion (DRG). Methods BMDMs were isolated and cultured, and lipopolysaccharide (LPS) combined with IFN-γ were used to induce M1 phenotype polarization of BMDMs, and then F4/80 and CD16/32 expression was detected by flow cytometry. The mature M1 type BMDMs were randomly divided into low-level laser group and control group. The laser exposure group was subjected to the laser treatments of 0.4J, 4J and 10J, and no laser was used in the control group. After 24 hours of laser exposure, the mRNA level of inducible nitric oxide synthase (iNOS) of M1 type BMDMs was detected by reverse transcription PCR, and the protein level of iNOS was detected by Western blot analysis. The levels of tumor necrosis factor alpha (TNF-α), interleukin-1ß (IL-1ß), brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in the supernatant of cultured cells were tested by ELISA. DRG neurons were cultured with the supernatant fluid of M1 type BMDMs, and immunofluorescence cytochemistry was employed to detect neuronal nuclei (NeuN) and ß-tubulin III expression of DRG neurons for determining the influence on the growth of DRG neuronal axons. Results Compared with the control group, the mRNA level of iNOS in M1 type BMDMs dramatically increased after 24 hours of low-level laser exposure. Among the 3 groups with different energy levels, the decrease of iNOS mRNA level was the most obvious in the group with 4J laser exposure. The protein levels of iNOS in the groups with 0.4J- and 4J- laser exposure were reduced more significantly than that in the control group, and the down-regulation was more prominent in the group with 4J laser exposure than that with 0.4J laser exposure. In addition, the secretion of TNF-α from M1 type BMDMs was reduced more significantly in the groups of 4J- and 10J- laser exposure than that in the control group. With regard to IL-1ß, its secretion was inhibited in all the laser exposure groups compared with the control group, and the suppression was more prominent in the groups of 0.4J- and 4J-laser exposure than that in the group of 10J-laser exposure. Furthermore, 4J-laser exposure significantly potentiated the secretion of BDNF and NGF in M1 type BMDMs compared with the control group. Moreover, co-culture with the supernatants from 4J- and 10J-laser exposure groups could significantly promote the growth of axons of DRG neurons. Conclusion Low-level laser exposure can inhibit the polarization of M1 type BMDM and the secretion of pro-inflammatory factor including TNF-α and IL-1ß. Besides, low-level laser exposure could contribute to the secretion of neurotrophic factors including BDNF and NGF, and promote the growth of DRG axon, and this effect is dose-dependent.


Assuntos
Axônios/fisiologia , Polaridade Celular , Gânglios Espinais/crescimento & desenvolvimento , Lasers , Macrófagos/citologia , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Células Cultivadas , Humanos , Interleucina-1beta/metabolismo , Lipopolissacarídeos , Macrófagos/efeitos da radiação , Fator de Crescimento Neural/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
20.
Artigo em Chinês | MEDLINE | ID: mdl-31189246

RESUMO

Objective: To Effects of n-hexane on learning and memory and the expressions of nerve growth factor (NGF) mRNA and nerve growth factor receptor (NGFR) mRNA of brain tissue in mice exposed to N-hexane. Methods: 40 Kunming mice were randomly divided into low-dose group, meddle-dose group, high-dose dose group and control group, with 10 mice in each group. All the groups were orally exposed to n-hexane in different doses: low-dose group with 43.5 mg/kg, middle-dose group with 86.5 mg/kg and high-dose group with 173.0 mg/kg, 1 time per day for 20 d. After the poisoning, the Y-arm test and the expressions of NGF mRNA and NGFR mRNA and the concentrations of NGF and NGFR in the brain tissues of each group were measured. Results: In the first Y-arm test, there existed a significant difference in correct reaction rate generally in all groups (P<0.05), and correct reaction rate in the middle-dose group and the high-dose group were low significantly compared with that in the control group(P<0.05). In the second Y-arm test, there existed a significant differences in total electric shock time and correct reaction rate generally in all groups (P<0.01), and the total electric shock time prolonged significantly and the correct response rate decreased significantly in 3 dose groups compared with those of the control group(P<0.05). The expression levels of NGF mRNA in brain tissues of low, meddle and high dose-groups were 0.81±0.66, 0.67±0.37 and 0.69±0.26, and the expression levels of NGFR mRNA were 1.22±0.42, 1.98±0.84 and 2.01±2.01, respectively. Compared with the control group, the expressions of NGF mRNA in the 3 dose groups decreased significantly (P<0.05), and the expression of NGFR mRNA in middle-and high-dose groups increased significantly (P<0.05). The concentrations of NGF in brain tissues of low,meddle and high dose-groups were 39.97±7.24 ng/L, 39.26±7.88 ng/L,31.70±8.21 ng/L,and the concentrations of NGFR were 17.37±6.82 ng/L,21.37±7.16 ng/L, 22.46±7.70 ng/L, respectively. Compared with the control group, the concentrations of NGF in high-dose groups decreased significantly(P<0.05), and the concentrations of NGFR in middle-and high-dose groups increased significantly (P<0.05). Conclusion: N-hexane exposure can result in decrease of learning and memory in mice, which may be related to abnormal expression of NGF mRNA and NGFR mRNA in brain tissue.


Assuntos
Hexanos , Aprendizagem , Memória , Fator de Crescimento Neural , Receptor de Fator de Crescimento Neural , Animais , Encéfalo , Hexanos/toxicidade , Aprendizagem/efeitos dos fármacos , Memória/efeitos dos fármacos , Camundongos , Fator de Crescimento Neural/efeitos dos fármacos , Fator de Crescimento Neural/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Distribuição Aleatória , Receptores de Fator de Crescimento Neural
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