RESUMO
The development of low-cost and effective natural products for treating neuron degenerative diseases have proven to be safe and potentially effective. Echium amoenum L. (Boraginaceae) is an annual herb that grows wildly in Europe and western Asia. The aim of this study was to evaluate the neuroprotective properties of an ethanol extract of E. amoenum L. The effects of E. amoenum L. extract on oxidative stress were measured in the rat R28 retinal precursor cell line. Furthermore, the protective role of the extract on the glutamate-induced and optic nerve crush (ONC) injury-induced cell death were evaluated in vitro and in vivo, respectively. Our results showed that the ethanol extract of E. amoenum L. prevented the glutamate-induced decrease in cell viability and increase in cell death in R28 cells and suppressed the overproduction of ROS induced by glutamate. Moreover, the extract significantly inhibited microglial activation and optic nerve damage induced by ONC injury in mice. In addition, the mechanism was attributed to the ability of the extract to decrease NF-κB pathway activation and its downstream inflammatory cytokine production. In conclusion, E. amoenum L. ethanol extract had a potent neuroprotective effect against glutamate-induced and ONC-induced cell death. This is likely due to its antioxidant and anti-inflammatory properties.
Assuntos
Produtos Biológicos , Lesões por Esmagamento , Echium , Fármacos Neuroprotetores , Traumatismos do Nervo Óptico , Animais , Antioxidantes/farmacologia , Produtos Biológicos/metabolismo , Produtos Biológicos/farmacologia , Sobrevivência Celular , Lesões por Esmagamento/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Etanol/farmacologia , Ácido Glutâmico/metabolismo , Camundongos , NF-kappa B/metabolismo , Fármacos Neuroprotetores/farmacologia , Nervo Óptico/metabolismo , Traumatismos do Nervo Óptico/tratamento farmacológico , Traumatismos do Nervo Óptico/metabolismo , Extratos Vegetais/metabolismo , Extratos Vegetais/farmacologia , Ratos , Espécies Reativas de Oxigênio/metabolismo , Células Ganglionares da Retina/metabolismoRESUMO
Choline is essential for maintaining the structure and function of cells in humans. Choline plays an important role in eye health and disease. It is a precursor of acetylcholine, a neurotransmitter of the parasympathetic nervous system, and it is involved in the production and secretion of tears by the lacrimal glands. It also contributes to the stability of the cells and tears on the ocular surface and is involved in retinal development and differentiation. Choline deficiency is associated with retinal hemorrhage, glaucoma, and dry eye syndrome. Choline supplementation may be effective for treating these diseases.
Assuntos
Colina/fisiologia , Oftalmopatias/metabolismo , Acetilcolina/biossíntese , Acetilcolina/fisiologia , Animais , Deficiência de Colina/complicações , Deficiência de Colina/fisiopatologia , Retinopatia Diabética/fisiopatologia , Síndromes do Olho Seco/tratamento farmacológico , Síndromes do Olho Seco/metabolismo , Síndromes do Olho Seco/fisiopatologia , Oftalmopatias/etiologia , Oftalmopatias/fisiopatologia , Dor Ocular/fisiopatologia , Glaucoma/fisiopatologia , Glicerilfosforilcolina/uso terapêutico , Humanos , Aparelho Lacrimal/inervação , Aparelho Lacrimal/metabolismo , Cristalino/metabolismo , Nociceptividade/fisiologia , Nervo Óptico/metabolismo , Sistema Nervoso Parassimpático/fisiopatologia , Fosfatidilcolinas/biossíntese , Fosfolipídeos/metabolismo , Receptores Nicotínicos/fisiologia , Retina/crescimento & desenvolvimento , Retina/metabolismo , Vasos Retinianos/metabolismo , Lágrimas/metabolismoRESUMO
Inflammatory demyelination and axonal injury of the optic nerve are hallmarks of optic neuritis (ON), which often occurs in multiple sclerosis and is a major cause of visual disturbance in young adults. Although a high dose of corticosteroids can promote visual recovery, it cannot prevent permanent neuronal damage. Novel and effective therapies are thus required. Given the recently defined capacity of matrine (MAT), a quinolizidine alkaloid derived from the herb Radix Sophorae flavescens, in immunomodulation and neuroprotection, we tested in this study the effect of matrine on rats with experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis. MAT administration, started at disease onset, significantly suppressed optic nerve infiltration and demyelination, with reduced numbers of Iba1+ macrophages/microglia and CD4+ T cells, compared to those from vehicle-treated rats. Increased expression of neurofilaments, an axon marker, reduced numbers of apoptosis in retinal ganglion cells (RGCs). Moreover, MAT treatment promoted Akt phosphorylation and shifted the Bcl-2/Bax ratio back towards an antiapoptotic one, which could be a mechanism for its therapeutic effect in the ON model. Taken as a whole, our results demonstrate that MAT attenuated inflammation, demyelination and axonal loss in the optic nerve, and protected RGCs from inflammation-induced cell death. MAT may therefore have potential as a novel treatment for this disease that may result in blindness.
Assuntos
Alcaloides/farmacologia , Apoptose/efeitos dos fármacos , Neurite Óptica/tratamento farmacológico , Quinolizinas/farmacologia , Células Ganglionares da Retina/efeitos dos fármacos , Animais , Axônios/efeitos dos fármacos , Axônios/metabolismo , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/metabolismo , Morte Celular/efeitos dos fármacos , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/tratamento farmacológico , Encefalomielite Autoimune Experimental/metabolismo , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Esclerose Múltipla/tratamento farmacológico , Esclerose Múltipla/metabolismo , Nervo Óptico/efeitos dos fármacos , Nervo Óptico/metabolismo , Neurite Óptica/metabolismo , Plantas Medicinais/química , Ratos , Ratos Wistar , Células Ganglionares da Retina/metabolismo , Transdução de Sinais/efeitos dos fármacos , MatrinasRESUMO
Considered to be one of the most important non-contagious systemic diseases worldwide, diabetes mellitus is still a topical issue on the health agenda with the problems it causes. Exposure to long-term hyperglycemia causes diabetic complications (diabetic neuropathy, nephropathy and retinopathy). The optic nerve can suffer damage by both diabetic retinopathy and neuropathy during diabetes, both because it is formed by axons of retinal ganglion cells and these axons belong to the central nervous system. The issue of hyperglycemia on the optic nerve have been described as diabetic papillopathy, posterior ischemic optic neuropathy, nonarteritic anterior ischemic optic neuropathy and optic atrophy in clinical studies. Experimental studies indicated axon-myelin degeneration in addition to microvascular and ultrastructural changes caused by the hyperglycemia-induced optic nerve damage. Although there are several proposed biochemical mechanisms to cause these damages, oxidative stress emerges as an important factor among them. Oxidative stress leads to pathological state on the nerve cells by affecting the DNA, protein and lipids at different levels. These are causing deterioration on nerve conduction velocity, myelin sheath and nerve structure, neurotrophic support system, glial cells and nerve function. Curcumin, as an important antioxidant, can be an ideal prophylactic agent to eliminate damages on optic nerve. Curcumin helps to regulate the balance of antioxidant and reactive oxygen species by targeting various molecules (NF-κB, STAT3, MAPK, Mfn2, Nrf2, pro-inflammatory cytokines). In addition, it shows healing or preventive effects on myelin sheath damage via regulating ferritin protein in oligodendrocytes. It is also effective in preventing neurovascular damage.
Assuntos
Curcumina/uso terapêutico , Hiperglicemia/complicações , Traumatismos do Nervo Óptico/tratamento farmacológico , Nervo Óptico/efeitos dos fármacos , Animais , Curcumina/farmacologia , Humanos , Hiperglicemia/metabolismo , Nervo Óptico/metabolismo , Traumatismos do Nervo Óptico/etiologia , Traumatismos do Nervo Óptico/metabolismo , Estresse Oxidativo/efeitos dos fármacosRESUMO
PURPOSE: Acute methanol exposure leads to systemic intoxication and toxic optic neuropathy. In this experimental study, we aimed to determine the protective effects of intravenous administration of ATP in methanol-induced optic neuropathy. MATERIALS AND METHODS: A total of 18 male albino Wistar rats weighing between 267 and 282 g were used for the experiment. The animals were divided into three groups as healthy control (HC), methanol (M), and methanol + ATP (M-ATP) groups. Distilled water was given to the healthy control group (n = 6) as the solvent, while 20% methanol was administered orally to the rats in M (n = 6) and M-ATP (n = 6) groups at a dose of 3 g/kg. Four hours after the administration of 20% methanol orally to the M-ATP group, ATP was injected intraperitoneally at a dose of 4 mg/kg. Eight hours after ATP injection, the animals were sacrificed by high-dose (50 mg/kg) thiopental anaesthesia and biochemical and histopathological examinations were performed on the removed optic nerve tissues. Malondialdehyde (MDA), total glutathione (tGSH), total oxidant status (TOS) and total anti-oxidant status (TAS) were analysed with biochemical tests. RESULTS: MDA, TOS and OSI were significantly higher and tGSH and TAS levels were significantly lower in methanol administered group compared with the healthy controls or M-ATP group (p: 0.001). There was not any significant difference between healthy controls and M-ATP group regarding the oxidative stress parameters. There was a significant destruction and increase in thickness and astrocyte numbers and edema-vacuolization in methanol administered group compared with the healthy controls or M-ATP group (p: 0.001). CONCLUSION: Intravenous ATP administration had a significant positive effect on the oxidative stress parameters and optic nerve structure in methanol-intoxicated rats. Antioxidant therapies should be considered in future studies as a possible therapy for methanol-induced toxic optic neuropathy.
Assuntos
Trifosfato de Adenosina/uso terapêutico , Antioxidantes/uso terapêutico , Traumatismos do Nervo Óptico/tratamento farmacológico , Trifosfato de Adenosina/farmacologia , Administração Intravenosa , Animais , Antioxidantes/farmacologia , Glutationa/metabolismo , Masculino , Malondialdeído/metabolismo , Metanol , Nervo Óptico/efeitos dos fármacos , Nervo Óptico/metabolismo , Nervo Óptico/patologia , Traumatismos do Nervo Óptico/induzido quimicamente , Traumatismos do Nervo Óptico/metabolismo , Traumatismos do Nervo Óptico/patologia , Estresse Oxidativo/efeitos dos fármacos , Ratos Wistar , SolventesRESUMO
Elevated intracranial pressure (ICP) can result in multiple neurologic sequelae including vision loss. Inducible models of ICP elevation are lacking in model organisms, which limits our understanding of the mechanism by which increased ICP impacts the visual system. We adapted a mouse model for the sustained elevation of ICP and tested the hypothesis that elevated ICP impacts the optic nerve and retinal ganglion cells (RGCs). ICP was elevated and maintained for 2 weeks, and resulted in multiple anatomic changes that are consistent with human disease including papilledema, loss of physiologic cupping, and engorgement of the optic nerve head. Elevated ICP caused a loss of RGC somas in the retina and RGC axons within the optic nerve, as well as a reduction in both RGC electrical function and contrast sensitivity. Elevated ICP also caused increased hypoxia-inducible factor (HIF)-1 alpha expression in the ganglion cell layer. These experiments confirm that sustained ICP elevation can be achieved in mice and causes phenotypes that preferentially impact RGCs and are similar to those seen in human disease. With this model, it is possible to model human diseases of elevated ICP such as Idiopathic Intracranial Hypertension and Spaceflight Associated Neuro-ocular Syndrome.
Assuntos
Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Hipertensão Intracraniana/complicações , Nervo Óptico/patologia , Células Ganglionares da Retina/patologia , Animais , Modelos Animais de Doenças , Eletrorretinografia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Hipertensão Intracraniana/diagnóstico por imagem , Hipertensão Intracraniana/metabolismo , Camundongos , Microscopia Eletrônica , Nervo Óptico/diagnóstico por imagem , Nervo Óptico/metabolismo , Fenótipo , Células Ganglionares da Retina/metabolismo , Tomografia de Coerência Óptica , Regulação para CimaRESUMO
OBJECTIVE: To study the protective effect of electroacupuncture (EA) along the visual conductive pathway for the optic nerve tissue of anterior ischemic optic neuropathy (AION) in terms of the structure and apoptosis. METHODS: The AION model of right eye was established with laser in 48 New Zealand white ear rabbits. All rabbits were randomly divided into a model group, an acupuncture group and an acupuncture combined with EA group, 16 rabbits in each one. Other 16 normal left eyes were selected as a blank group. Acupuncture and EA of 30 min were used in the corresponding groups for 3 days at the right "Cuanzhu" (BL 2), "Yuyao" (EX-HN 4) and "Qiaomingxue" (Extra), once a day. There was no intervention in the model group and the blank group. The morphological structure and retinal thickness of lining of the optic nerve were detected with HE stain. The expressions of the Bcl-2 and Bax in the retina were detected with immunohistochemistry. And the concentration of tumor necrosis factor-α (TNF-α) in the retina was detected with enzyme linked immunosorbent assay (ELISA). RESULTS: In the model group, the ganglion cell layer revealed hyperplasia and disorder, and the retina ganglion cells revealed loose arrangements, thin inner nuclear layers, and reduction of cell amounts, some long parts missing cells. However, the above structural damages were much weaker in the acupuncture group and acupuncture combined with EA group. The inner layer of the retina in the model group was thinner than that in the blank group (P<0.05). That in the acupuncture combined with EA group showed significant better results than those in the acupuncture and model groups (both P<0.05), which was not statistically different from that in the normal group (P>0.05). The Bcl-2 count in the model group was lower than that in the blank group (P<0.05), and that in the acupuncture combined with EA group was better than those in the acupuncture and model groups (both P<0.05), not significantly different from that in the blank group (P>0.05). The number of Bax in the model group was higher than that in the blank group (P<0.05), and that in the acupuncture combined with EA group was lower than those in the acupuncture and model groups (both P<0.05), and was similar to that in the blank group (P>0.05). Bcl-2/Bax in the model group was lower than that in the blank group (P<0.05). The value in the acupuncture combined with EA group presented better than those in the acupuncture and model groups (both P<0.05), which had no difference from that in the blank group (P>0.05). TNF-α in the model group was higher than that in the blank group (P<0.05), and no such differences were detected between other groups and the model group (bothP>0.05). CONCLUSIONS: EA along the visual conductive pathway is protective to some extent for optic nerve tissue, which can increase the expression of Bcl-2 and reduce the expression of Bax so as to restrain ganglion cell apoptosis.
Assuntos
Apoptose , Eletroacupuntura/métodos , Nervo Óptico/patologia , Neuropatia Óptica Isquêmica/terapia , Vias Visuais , Pontos de Acupuntura , Terapia por Acupuntura/métodos , Animais , Hiperplasia/diagnóstico , Nervo Óptico/metabolismo , Neuropatia Óptica Isquêmica/metabolismo , Neuropatia Óptica Isquêmica/patologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Coelhos , Distribuição Aleatória , Retina/química , Fator de Necrose Tumoral alfa/análise , Proteína X Associada a bcl-2/metabolismoRESUMO
AIMS: Optic neuritis (ON) is an inflammatory disease of the optic nerve, which often occurs in patients with multiple sclerosis (MS) and leads to retinal ganglion cell (RGC) death and even severe visual loss. Valproic acid (VPA) is a short-chain branched fatty acid with anti-epileptic, neuro-protective and anti-inflammatory effects. Here, we examined the effects of VPA in experimental autoimmune encephalomyelitis (EAE) rats and explored the underlying mechanisms. MAIN METHODS: EAE was induced by subcutaneous injection with myelin basic protein, emulsified with complete Freund's adjuvant and Mycobacterium tuberculosis H37Ra into the Lewis rats. Subsequently, animals in the VPA groups were treated orally with VPA (250 or 500 mg/kg) once a day for 13 days. KEY FINDINGS: VPA treatment significantly attenuated inflammation and microgliosis in optic nerve in EAE-ON rats, as evidenced by the decrease in the mRNA levels of interferon (INF)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-17, and inducible nitric oxide synthase (iNOS), the suppression in nuclear factor (NF)-κB signal pathway as well as the down-regulation of CD11b expression in optic nerve. Additionally, the apoptotic RGCs were remarkably increased in the EAE retina, which was inhibited by VPA treatment. Consistent with the TUNEL staining, VPA administration also obviously suppressed the ratio of Bax: Bcl-2 and the expression of cleaved caspase-3 and PARP in optic nerve in EAE rats. SIGNIFICANCE: Our findings demonstrated that VPA treatment could prevent inflammation responses and RGC apoptosis in optic nerve in EAE-ON rats, suggesting that VPA may be available for optic nerve protection during ON.
Assuntos
Apoptose/efeitos dos fármacos , Inflamação/tratamento farmacológico , Nervo Óptico/efeitos dos fármacos , Neurite Óptica/tratamento farmacológico , Células Ganglionares da Retina/efeitos dos fármacos , Ácido Valproico/farmacologia , Animais , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/tratamento farmacológico , Encefalomielite Autoimune Experimental/metabolismo , Marcação In Situ das Extremidades Cortadas/métodos , Inflamação/metabolismo , Interferon gama/metabolismo , Interleucina-17/metabolismo , Interleucina-1beta/metabolismo , Masculino , NF-kappa B/metabolismo , Nervo Óptico/metabolismo , Neurite Óptica/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos Lew , Células Ganglionares da Retina/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The northern elephant seal (Mirounga angustirostris) and California sea lion (Zalophus californianus) are members of a diverse clade of carnivorous mammals known as pinnipeds. Pinnipeds are notable for their large, ape-sized brains, yet little is known about their central nervous system. Both the northern elephant seal and California sea lion spend most of their lives at sea, but each also spends time on land to breed and give birth. These unique coastal niches may be reflected in specific evolutionary adaptations to their sensory systems. Here, we report on components of the visual pathway in these two species. We found evidence for two classes of myelinated fibers within the pinniped optic nerve, those with thick myelin sheaths (elephant seal: 9%, sea lion: 7%) and thin myelin sheaths (elephant seal: 91%, sea lion: 93%). In order to investigate the architecture of the lateral geniculate nucleus, superior colliculus, and primary visual cortex, we processed brain sections from seal and sea lion pups for Nissl substance, cytochrome oxidase, and vesicular glutamate transporters. As in other carnivores, the dorsal lateral geniculate nucleus consisted of three main layers, A, A1, and C, while each superior colliculus similarly consisted of seven distinct layers. The sea lion visual cortex is located at the posterior side of cortex between the upper and lower banks of the postlateral sulcus, while the elephant seal visual cortex extends far more anteriorly along the dorsal surface and medial wall. These results are relevant to comparative studies related to the evolution of large brains.
Assuntos
Nervo Óptico/anatomia & histologia , Leões-Marinhos/anatomia & histologia , Focas Verdadeiras/anatomia & histologia , Colículos Superiores/anatomia & histologia , Tálamo/anatomia & histologia , Córtex Visual/anatomia & histologia , Animais , Animais Recém-Nascidos , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Feminino , Masculino , Nervo Óptico/metabolismo , Colículos Superiores/metabolismo , Tálamo/metabolismo , Proteína Vesicular 1 de Transporte de Glutamato/metabolismo , Proteína Vesicular 2 de Transporte de Glutamato/metabolismo , Córtex Visual/metabolismo , Vias Visuais/anatomia & histologia , Vias Visuais/metabolismoRESUMO
Iron deficiency has a critical impact on maturational mechanisms of the brain and the damage related to neuroanatomical parameters is not satisfactorily reversed after iron replacement. However, emerging evidence suggest that enriched early experience may offer great therapeutic efficacy in cases of nutritional disorders postnatally, since the brain is remarkably responsive to its interaction with the environment. Given the fact that tactile stimulation (TS) treatment has been previously shown to be an effective therapeutic approach and with potential application to humans, here we ask whether exposure to TS treatment, from postnatal day (P) 1 to P32 for 3min/day, could also be employed to prevent neuroanatomical changes in the optic nerve of rats maintained on an iron-deficient diet during brain development. We found that iron deficiency changed astrocyte, oligodendrocyte, damaged fiber, and myelinated fiber density, however, TS reversed the iron-deficiency-induced alteration in oligodendrocyte, damaged fiber and myelinated fiber density, but failed to reverse astrocyte density. Our results suggest that early iron deficiency may act by disrupting the timing of key steps in visual system development thereby modifying the normal progression of optic nerve maturation. However, optic nerve development is sensitive to enriching experiences, and in the current study we show that this sensitivity can be used to prevent damage from postnatal iron deficiency during the critical period.
Assuntos
Deficiências de Ferro , Manipulações Musculoesqueléticas , Nervo Óptico/crescimento & desenvolvimento , Vias Visuais/crescimento & desenvolvimento , Animais , Animais Recém-Nascidos , Astrócitos/metabolismo , Astrócitos/patologia , Peso Corporal , Dieta , Modelos Animais de Doenças , Manobra Psicológica , Masculino , Fibras Nervosas Mielinizadas/metabolismo , Fibras Nervosas Mielinizadas/patologia , Neuroproteção , Oligodendroglia/metabolismo , Oligodendroglia/patologia , Nervo Óptico/irrigação sanguínea , Nervo Óptico/metabolismo , Nervo Óptico/patologia , Estimulação Física , Distribuição Aleatória , Ratos Wistar , Vias Visuais/irrigação sanguínea , Vias Visuais/metabolismo , Vias Visuais/patologiaRESUMO
Glutamate excitotoxicity plays a major role in the loss of retinal ganglion cells (RGCs) in glaucoma. The toxic effects of glutamate on RGCs are mediated by the overstimulation of N-methyl-D-aspartate (NMDA) receptors. Accordingly, NMDA receptor antagonists have been suggested to inhibit excitotoxicity in RGCs and delay the progression and visual loss in glaucoma patients. The purpose of the present study was to examine the potential neuroprotective effect of Mg acetyltaurate (MgAT) on RGC death induced by NMDA. MgAT was proposed mainly due to the combination of magnesium (Mg) and taurine which may provide neuroprotection by dual mechanisms of action, i.e., inhibition of NMDA receptors and antioxidant effects. Rats were divided into 5 groups and were given intravitreal injections. Group 1 (PBS group) was injected with vehicle; group 2 (NMDA group) was injected with NMDA while groups 3 (pre-), 4 (co-), and 5 (post-) treatments were injected with MgAT, 24 h before, in combination or 24 h after NMDA injection respectively. NMDA and MgAT were injected in PBS at doses 160 and 320 nmol, respectively. Seven days after intravitreal injection, the histological changes in the retina were evaluated using hematoxylin & eosin (H&E) staining. Optic nerves were dissected and stained in Toluidine blue for grading on morphological neurodegenerative changes. The extent of apoptosis in retinal tissue was assessed by TUNEL assay and caspase-3 immunohistochemistry staining. The estimation of neurotrophic factor, oxidative stress, pro/anti-apoptotic factors and caspase-3 activity in retina was done using enzyme-linked immunosorbent assay (ELISA) technique. The retinal morphometry showed reduced thickness of ganglion cell layer (GCL) and reduction in the number of retinal cells in GCL in NMDA group compared to the MgAT-treated groups. TUNEL and caspase-3 staining showed increased number of apoptotic cells in inner retina. The results were further corroborated by the estimation of neurotrophic factor, oxidative stress, pro/anti-apoptotic factors, and caspase-3 activity in retina. In conclusion, current study revealed that intravitreal MgAT prevents retinal and optic nerve damage induced by NMDA. Overall, our data demonstrated that the pretreatment with MgAT was more effective than co- and posttreatment. This protective effect of MgAT against NMDA-induced retinal cell apoptosis could be attributed to the reduction of retinal oxidative stress and activation of BDNF-related neuroprotective mechanisms.
Assuntos
N-Metilaspartato/toxicidade , Fármacos Neuroprotetores/farmacologia , Células Ganglionares da Retina/efeitos dos fármacos , Taurina/análogos & derivados , Animais , Apoptose/efeitos dos fármacos , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Avaliação Pré-Clínica de Medicamentos , Agonistas de Aminoácidos Excitatórios/toxicidade , Feminino , Injeções Intravítreas , Masculino , Nervo Óptico/efeitos dos fármacos , Nervo Óptico/metabolismo , Nervo Óptico/patologia , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Distribuição Aleatória , Ratos Sprague-Dawley , Células Ganglionares da Retina/metabolismo , Células Ganglionares da Retina/patologia , Taurina/farmacologia , Fatores de TempoRESUMO
Injury to the central nervous system causes progressive degeneration of injured axons, leading to loss of the neuronal bodies. Neuronal survival after injury is a prerequisite for successful regeneration of injured axons. In this study, we investigated the effects of increased production of omega-3 fatty acids and elevation of cAMP on retinal ganglion cell (RGC) survival and axonal regeneration after optic nerve (ON) crush injury in adult mice. We found that increased production of omega-3 fatty acids in mice enhanced RGC survival, but not axonal regeneration, over a period of 3 weeks after ON injury. cAMP elevation promoted RGC survival in wild type mice, but no significant difference in cell survival was seen in mice over-producing omega-3 fatty acids and receiving intravitreal injections of CPT-cAMP, suggesting that cAMP elevation protects RGCs after injury but does not potentiate the actions of the omega-3 fatty acids. The observed omega-3 fatty acid-mediated neuroprotection is likely achieved partially through ERK1/2 signaling as inhibition of this pathway by PD98059 hindered, but did not completely block, RGC protection. Our study thus enhances our current understanding of neural repair after CNS injury, including the visual system.
Assuntos
Ácidos Graxos Ômega-3/metabolismo , Traumatismos do Nervo Óptico/metabolismo , Células Ganglionares da Retina/metabolismo , Análise de Variância , Animais , Axônios/metabolismo , Contagem de Células , Sobrevivência Celular , Proteínas Quinases Dependentes de AMP Cíclico/fisiologia , Modelos Animais de Doenças , Sistema de Sinalização das MAP Quinases/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Compressão Nervosa , Regeneração Nervosa/fisiologia , Nervo Óptico/metabolismo , Traumatismos do Nervo Óptico/patologia , Células Ganglionares da Retina/citologiaRESUMO
Reactive oxygen species (ROS) are implicated in a range of degenerative conditions, including aging, neurodegenerative diseases, and neurological disorders. Myelin is a lipid-rich multilamellar sheath that facilitates rapid nerve conduction in vertebrates. Given the high energetic demands and low antioxidant capacity of the cells that elaborate the sheaths, myelin is considered intrinsically vulnerable to oxidative damage, raising the question whether additional mechanisms prevent structural damage. We characterized the structural and biochemical basis of ROS-mediated myelin damage in murine tissues from both central nervous system (CNS) and peripheral nervous system (PNS). To determine whether ROS can cause structural damage to the internodal myelin, whole sciatic and optic nerves were incubated ex vivo with a hydroxyl radical-generating system consisting of copper (Cu), hydrogen peroxide (HP), and ortho-phenanthroline (OP). Quantitative assessment of unfixed tissue by X-ray diffraction revealed irreversible compaction of myelin membrane stacking in both sciatic and optic nerves. Incubation in the presence of the hydroxyl radical scavenger sodium formate prevented this damage, implicating hydroxyl radical species. Myelin membranes are particularly enriched in plasmalogens, a class of ether-linked phospholipids proposed to have antioxidant properties. Myelin in sciatic nerve from plasmalogen-deficient (Pex7 knockout) mice was significantly more vulnerable to Cu/OP/HP-mediated ROS-induced compaction than myelin from WT mice. Our results directly support the role of plasmalogens as endogenous antioxidants providing a defense that protects ROS-vulnerable myelin.
Assuntos
Sequestradores de Radicais Livres/farmacologia , Bainha de Mielina/metabolismo , Plasmalogênios/farmacologia , Animais , Quelantes/farmacologia , Avaliação Pré-Clínica de Medicamentos , Ácido Edético/farmacologia , Formiatos/farmacologia , Camundongos Knockout , Bainha de Mielina/efeitos dos fármacos , Nervo Óptico/metabolismo , Nervo Óptico/patologia , Oxirredução , Estresse Oxidativo , Receptor 2 de Sinal de Orientação para Peroxissomos , Carbonilação Proteica , Espécies Reativas de Oxigênio/metabolismo , Receptores Citoplasmáticos e Nucleares/genética , Nervo Isquiático/metabolismo , Nervo Isquiático/patologiaRESUMO
Glycogen is present in the mammalian nervous system, but at concentrations of up to one hundred times lower than those found in liver and skeletal muscle. This relatively low concentration has resulted in neglect of assigning a role(s) for brain glycogen, but in the last 15 years enormous progress has been made in revealing the multifaceted roles that glycogen plays in the mammalian nervous system. Initial studies highlighted a role for glycogen in supporting neural elements (neurons and axons) during aglycemia, where glycogen supplied supplementary energy substrate in the form of lactate to fuel neural oxidative metabolism. The appropriate enzymes and membrane bound transporters have been localized to cellular locations consistent with astrocyte to neuron energy substrate shuttling. A role for glycogen in supporting the induction of long term potential (LTP) in the hippocampus has recently been described, where glycogen is metabolized to lactate and shuttled to neurons via the extracellular space by monocarboxylate transporters, where it plays an integral role in the induction process of LTP. This is the first time that glycogen has been assigned a role in a distinct, complex physiological brain function, where the lack of glycogen, in the presence of normoglycemia, results in disturbance of the function. The signalling pathway that alerts astrocytes to increased neuronal activity has been recently described, highlighting a pivotal role for increased extracellular potassium ([K(+)]o) that routinely accompanies increased neural activity. An astrocyte membrane bound bicarbonate transporter is activated by the [K(+)]o, the resulting increase in intracellular bicarbonate alkalizing the cell's interior and activating soluble adenyl cyclase (sAC). The sAC promotes glycogenolysis via increases in cyclic AMP, ultimately producing lactate, which is shuttled out of the astrocyte and presumably taken up by neurons from the extracellular space.
Assuntos
Sistema Nervoso Central/metabolismo , Metabolismo Energético , Glicogênio/metabolismo , Sistema Nervoso Periférico/metabolismo , Animais , Astrócitos/classificação , Astrócitos/metabolismo , Encéfalo/citologia , Encéfalo/metabolismo , Glucose/metabolismo , Hipocampo/metabolismo , Humanos , Lactatos/metabolismo , Potenciação de Longa Duração/fisiologia , Camundongos , Modelos Neurológicos , Neurônios/metabolismo , Nervo Óptico/metabolismo , Especificidade de Órgãos , Potássio/metabolismo , Ratos , Transdução de Sinais , ATPase Trocadora de Sódio-Potássio/metabolismo , Transmissão Sináptica/fisiologiaRESUMO
To investigate the effects of cerebrolysin (Cbl) on optic nerves (ON) and retinal ganglion cells (RGC) in a rat model of ON crush. Rats received intravitreal injection of Cbl (n = 20), intra-ON injection of Cbl (n = 20), intraperitoneal injection (IPI) of Cbl (n = 20), or phosphate buffered saline (PBS; n = 20) every day for 2 weeks after ON crush injury. At 3 weeks post-trauma, RGC density was counted by retrograde labeling with FluoroGold and visual function was assessed by flash visual-evoked potentials. Activities of microglia after insults were quantified by immunohistochemical analysis of the presence of ED1 in the optic nerve. At 3 weeks postcrush, the densities of RGCs in the Cbl-IVI group (1125 ± 166/mm(2)) and in the Cbl-IPI treatment group (1328 ± 119/mm(2)) were significantly higher than those in the PBS group (641 ± 214/mm(2)). The flash visual-evoked potential measurements showed that latency of the P1 wave was significantly shorter in the Cbl-IVI- and Cbl-IPI-treated groups (105 ± 4 ms and 118 ± 26 ms, respectively) than in the PBS-treated group (170 ± 20 ms). However, only Cbl IPI treatment resulted in a significant decrease in the number of ED1-positive cells at the lesion sites of the ON (5 ± 2 cells/vs. 30 ± 4 cells/high-power field in control eyes). Treatment with intra-ON injection of Cbl was harmful to the optic nerve in the crush model. Systemic administration of Cbl had neuroprotective effects on RGC survival and visual function in the optic nerve crush model.
Assuntos
Aminoácidos/farmacologia , Fármacos Neuroprotetores/farmacologia , Traumatismos do Nervo Óptico/tratamento farmacológico , Animais , Avaliação Pré-Clínica de Medicamentos , Potenciais Evocados Visuais , Masculino , Nervo Óptico/efeitos dos fármacos , Nervo Óptico/metabolismo , Nervo Óptico/fisiopatologia , Ratos WistarRESUMO
BACKGROUND: beta-elemene is a natural antitumor plant drug. Beneficial effects of beta-elemene therapy have been demonstrated in some kinds of tumor clinically. Especially, it has been found to pass through the blood brain barrier easily. Other reports have indicated that immune disorder that appeared in some tumors usually can be seen in demyelinating diseases including multiple sclerosis and experimental autoimmune encephalomyelitis. However, no information regarding the effects of beta-elemene therapy on the T helper cell subsets, Th1 or Th17 cells in experimental autoimmune encephalomyelitis has been found. METHODS AND FINDINGS: We first determined morphologically that beta-elemene therapy markedly suppressed the inflammation in experimental autoimmune encephalomyelitis optic nerve. We then determined the effect in vivo of beta-elemene on Treg cells and Th17 and Th1 cells. We found that beta-elemene treatment modulated immune balance in both the periphery and the inflamed optic nerve by promoting less downregulation in Treg cells, inhibiting Th17 and Th1 polarization. CONCLUSIONS: Taken together, our finding reveals an important new locus where beta-elemene induces substantial protection in experimental autoimmune encephalomyelitis optic nerve through signaling to several critical subsets of immune cells that reside in the peripheral and central nervous system.
Assuntos
Encefalomielite Autoimune Experimental/tratamento farmacológico , Nervo Óptico/efeitos dos fármacos , Fitoterapia , Extratos Vegetais/administração & dosagem , Sesquiterpenos/administração & dosagem , Animais , Progressão da Doença , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/fisiopatologia , Feminino , Glicoproteínas/administração & dosagem , Glicoproteínas/imunologia , Humanos , Interferon gama/genética , Interferon gama/metabolismo , Interleucina-17/genética , Interleucina-17/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Esclerose Múltipla , Glicoproteína Mielina-Oligodendrócito , Nervo Óptico/metabolismo , Nervo Óptico/patologia , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/imunologiaRESUMO
BACKGROUND: The optic nerve is a pure white matter central nervous system (CNS) tract with an isolated blood supply, and is widely used in physiological studies of white matter response to various insults. We examined the gene expression profile of human optic nerve (ON) and, through the NEIBANK online resource, to provide a resource of sequenced verified cDNA clones. An un-normalized cDNA library was constructed from pooled human ON tissues and was used in expressed sequence tag (EST) analysis. Location of an abundant oligodendrocyte marker was examined by immunofluorescence. Quantitative real time polymerase chain reaction (qRT-PCR) and Western analysis were used to compare levels of expression for key calcium channel protein genes and protein product in primate and rodent ON. RESULTS: Our analyses revealed a profile similar in many respects to other white matter related tissues, but significantly different from previously available ON cDNA libraries. The previous libraries were found to include specific markers for other eye tissues, suggesting contamination. Immune/inflammatory markers were abundant in the new ON library. The oligodendrocyte marker QKI was abundant at the EST level. Immunofluorescence revealed that this protein is a useful oligodendrocyte cell-type marker in rodent and primate ONs. L-type calcium channel EST abundance was found to be particularly low. A qRT-PCR-based comparative mammalian species analysis reveals that L-type calcium channel expression levels are significantly lower in primate than in rodent ON, which may help account for the class-specific difference in responsiveness to calcium channel blocking agents. Several known eye disease genes are abundantly expressed in ON. Many genes associated with normal axonal function, mRNAs associated with axonal transport, inflammation and neuroprotection are observed. CONCLUSION: We conclude that the new cDNA library is a faithful representation of human ON and EST data provide an initial overview of gene expression patterns in this tissue. The data provide clues for tissue-specific and species-specific properties of human ON that will help in design of therapeutic models.
Assuntos
Axônios/metabolismo , DNA Complementar/metabolismo , Etiquetas de Sequências Expressas/metabolismo , Oligodendroglia/metabolismo , Nervo Óptico/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Northern Blotting , Western Blotting , DNA Complementar/análise , DNA Complementar/genética , Bases de Dados Genéticas , Feminino , Imunofluorescência , Perfilação da Expressão Gênica , Biblioteca Gênica , Humanos , Macaca , Masculino , Microscopia Confocal , Pessoa de Meia-Idade , Nervo Óptico/química , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE) are characterized by T cell-mediated autoimmune inflammation of the central nervous system (CNS) leading to oligodendrocyte loss and demyelination accompanied by neuronal cell death. Neuronal TWIK-related acid-sensitive potassium (TASK) channels allow the regulated efflux of potassium ions. These channels might either protect neurons in the inflamed CNS by modulating electrical excitability or even contribute to inflammatory neurodegeneration mediating intracellular potassium depletion. Using a combination of in-situ-hybridisation and immunofluorescence staining, we found increased neuronal expression of TASK1 and TASK3 channels in the optic nerve and decreased expression in the spinal cord and thalamus of rats undergoing MOG-induced EAE. Inflammatory plaques of human MS patients displayed profoundly lowered expression of both TASK isoforms. Thus, regulated expression of TASK channels might contribute to a molecular switch between death and survival of neurons in autoimmune CNS inflammation.
Assuntos
Sistema Nervoso Central/metabolismo , Encefalomielite Autoimune Experimental/metabolismo , Esclerose Múltipla/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Canais de Potássio de Domínios Poros em Tandem/metabolismo , Animais , Sobrevivência Celular/genética , Sobrevivência Celular/imunologia , Sistema Nervoso Central/imunologia , Sistema Nervoso Central/fisiopatologia , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/fisiopatologia , Imunofluorescência , Humanos , Hibridização In Situ , Esclerose Múltipla/imunologia , Esclerose Múltipla/fisiopatologia , Proteínas do Tecido Nervoso/genética , Neurônios/imunologia , Nervo Óptico/imunologia , Nervo Óptico/metabolismo , Nervo Óptico/patologia , Potássio/metabolismo , Canais de Potássio de Domínios Poros em Tandem/genética , Ratos , Medula Espinal/imunologia , Medula Espinal/metabolismo , Medula Espinal/patologia , Tálamo/imunologia , Tálamo/metabolismo , Tálamo/patologiaRESUMO
PURPOSE: To evaluate the efficacy of intravitreal injection of GDNF-loaded biodegradable microspheres in promoting the survival of retinal ganglion cells (RGCs) and their axons in a rat model of chronically elevated intraocular pressure (IOP). METHODS: Chronic elevation of IOP was induced in Brown Norway rats through injection of hypertonic saline (1.9 M) into the episcleral veins. After injection, IOP was measured twice a week in rats using topical anesthesia. Poly DL-lactide-co-glycolide (PLGA) microspheres were fabricated using a modified version of the spontaneous emulsification technique. Two and ten percent of volume solutions of microspheres loaded with glial cell line-derived neurotrophic factor (GDNF) were injected into the vitreous cavity of rats with elevated IOP, with injections of blank microspheres and PBS serving as controls. Histological analysis was used to quantify surviving RGCs and axons and provide comparison among different groups. In addition, the thickness of the retinal inner plexiform layer (IPL) and the level of glial fibrillary acidic protein (GFAP) expression within the retina and optic nerve were quantitatively compared. RESULTS: IOP was significantly increased in eyes with episcleral vein injection over untreated eyes (p<0.001) but did not show a significant difference among groups that received intravitreal injections of GDNF microspheres, blank microspheres, or PBS (p=0.1852). The duration of IOP elevation in this experiment was eight weeks. Expression of GDNF and its receptors localizes to the adult rat RGCs. Ten percent of the GDNF microsphere treatment significantly increased RGC survival and axon survival (p<0.001), reduced the loss of retinal IPL thickness (p<0.001), and decreased glial cell activation in the retina and optic nerve (p<0.001) compared with blank microspheres and PBS. In addition, GDNF microsphere treatment moderately reduced cupping of the optic nerve head. CONCLUSIONS: Delivery of GDNF via biodegradable microspheres significantly increased the survival of RGCs and their axons, preserved IPL thickness, and decreased retina and optic nerve glial cell activation in an experimental glaucoma model. This study suggests that GDNF delivered by PLGA microspheres may be useful as a neuroprotective tool in the treatment of glaucomatous optic neuropathy.
Assuntos
Glaucoma/fisiopatologia , Fator Neurotrófico Derivado de Linhagem de Célula Glial/administração & dosagem , Fármacos Neuroprotetores/administração & dosagem , Nervo Óptico/fisiopatologia , Retina/fisiopatologia , Implantes Absorvíveis , Animais , Axônios/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Glaucoma/patologia , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/farmacologia , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Injeções , Pressão Intraocular/efeitos dos fármacos , Microesferas , Fármacos Neuroprotetores/farmacologia , Nervo Óptico/efeitos dos fármacos , Nervo Óptico/metabolismo , Proteínas Proto-Oncogênicas c-ret/metabolismo , Ratos , Ratos Endogâmicos BN , Retina/efeitos dos fármacos , Retina/metabolismo , Retina/patologia , Células Ganglionares da Retina/efeitos dos fármacos , Células Ganglionares da Retina/patologia , Solução Salina Hipertônica/administração & dosagem , Solução Salina Hipertônica/farmacologia , Corpo VítreoRESUMO
Unlike mammals, the fish optic nerve can regenerate after injury. So far, many growth or trophic factors have been shown as an axon-regenerating molecule. However, it is totally unknown what substance regulates or triggers the activity of these factors on axonal elongation. Therefore, we constructed a goldfish retina cDNA library prepared from the retina treated with optic nerve transection 5 d previously, when it was just before regrowing optic axons after injury. A cDNA clone for goldfish purpurin for which expression was upregulated during the early stage of optic nerve regeneration was isolated from the retina cDNA library. Purpurin was discovered as a secretory retinol-binding protein in developing chicken retinas. Levels of purpurin mRNA and protein transiently increased and rapidly decreased 2-5 d and 10 d after axotomy, respectively. Purpurin mRNA was localized to the photoreceptor cells, whereas the protein was diffusely found in all of the retinal layers. A recombinant purpurin alone did not affect any change of neurite outgrowth in explant culture of the control retina, whereas a concomitant addition of the recombinant purpurin and retinol first induced a drastic enhancement of neurite outgrowth. Furthermore, the action of retinol-bound purpurin was effective only in the control (untreated) retinas but not in those primed (treated) with a previous optic nerve transection. Thus, purpurin with retinol is the first candidate molecule of priming neurite outgrowth in the early stage of optic nerve regeneration in fish.