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Objective To study the effect of matrine on Calpain and MAP-2 in rats with experimental autoimmune encephalomyelitis(EAE).Methods In accordance with the random number table ,60 Wistar rats were divided into 6 groups randomly: normal group,model group,dexamethasone(DEX)-treated group(1mg/kg),high-dose matrine(MAT)-treated group(250mg/kg),middle-dose MAT-treated group(200mg/kg) and low-dose MAT-treated group(150mg/kg).The EAE models were induced by immunized spinal cord extracts of guinea pig with complete Freunds'adjuvant.Rats of three MAT-treated groups and DEX-treated group were injected intraper-itoneally with MAT and DEX daily for 16 days respectively,whereas rats of normal group and model group were injected intraperitoneally with normal saline.Clinical signs of rats in six groups were observed daily .Hematoxylin-eosin (HE) was used to analyze histopathological evaluation of spinal cord .μ-Calpain,m-Calpain and MAP-2 in spinal cord were determined using RT-PCR and immunohistochemistry respectively .Results Compared with the model group[(2.85 ±0.78)points],the clinical scores were significantly decreased in high-dose-MAT group[(1.28 ± 0.59) points], middle-dose-MAT group [(1.45 ±0.64) points] and low-dose-MAT group [(2.09 ± 0.71)points](t =5.345,4.314,2.869,all P <0.05).The HE score of rats in model group[(2.49 ±0.29)points] was significantly higher than that in high-dose-MAT group[(1.04 ±0.26) points],middle-dose-MAT group [(1.29 ±0.20) points] and low-dose-MAT group[(1.77 ±0.24)points] (t =5.185,4.274,3.629,all P <0.01).The levels of μ-Calpain mRNA and m-Calpain mRNA in the three MAT-treated groups were significantly lower than those in model group(t =10.656,9.418,7.044,all P <0.01;t =6.332,5.416,3.978,all P <0.01).In addition,the expression of MAP-2 in the spinal cord of EAE rats showed a marked elevation after MAT treatment (t =12.841,9.924,7.038,all P <0.01).Conclusion Matrine may be an effective therapeutic approach for EAE by inhibiting Calpain and increase MAP-2 expression.
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Purpose Cortex is one of the frequently involved sites of multiple sclerosis (MS),and the cortex and corpus callosum lesions of MS are gradually concerned.The study aims to observe the changes of cerebral cortex and corpus callosum of MS in experimental autoimmune encephalomyelitis (EAE) model by using 7.0T MRI diffusion tensor imaging (DTI).Materials and Methods Twenty female C57BL/6 mice of 6-8 week old were enrolled in the study,10 of which were induced by MOG35-55 to make EAE models and the rest 10 of which were taken as control group.On the 20 days after model establishment,the head T2WI and DTI were performed on both control and EAE mice.DTI quantitative indicators such as fractional anisotropy (FA),mean diffusivity (MD),axial dispersion coefficient λ//,and radial dispersion coefficient λ ⊥ in region of interest including bilateral prefrontal cortex,bilateral cingulate cortex and corpus callosum were compared between the two groups.Results No obvious lesions were observed on the T2WI in both control and EAE groups.In the experimental group,the FA mapping suggested the integrity of the left side of the corpus callosum was destroyed.The FA,MD,λ// λ ⊥ of bilateral prefrontal cortex and corpus callosum showed significant difference between experimental group and control group (P＜0.05);the increase of λ ⊥ in bilateral cingulate was significantly different from that in the control group.Meanwhile,HE staining in the experimental group showed that inflammatory cells gathered around the cortical and subcortical vessels.The LFB staining in experimental group showed a bit paler than that in the control group,and the corpus callosum showed patchy demyelination.Conclusion The technique of 7.0T MRI DTI sequence can detect cortex and corpus callosum lesions which cannot be found by conventional MRI,so that it provides radiological evidence for the study of MS with cortex and corpus callosum lesions.
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PURPOSE: Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system. It has been shown that memory deficits is common in patients with MS. Recent studies using experimental autoimmune encephalomyelitis (EAE) as an animal model of MS have shown that indicated that EAE causes hippocampal-dependent impairment in learning and memory. Thus far, there have been no in vivo electrophysiological reports describing synaptic transmission in EAE animals. The aim of the present work is to evaluate the synaptic changes in the CA1 region of the hippocampus of EAE rats. METHODS: To evaluate changes in synaptic transmission in the CA1 region of the hippocampus of EAE rats, field excitatory postsynaptic potentials (fEPSPs) from the stratum radiatum of CA1 neurons, were recorded following Schaffer collateral stimulation. RESULTS: The results showed that EAE causes deficits in synaptic transmission and long-term potentiation (LTP) in the hippocampus. In addition, paired-pulse index with a 120 msec interstimulus interval was decreased in the EAE group. These findings indicate that EAE might induce suppression in synaptic transmission and LTP by increasing the inhibitory effect of GABAB receptors on the glutamate-mediated EPSP. CONCLUSIONS: In conclusion, influence of inflammation-triggered mechanisms on synaptic transmission may explain the negative effect of EAE on learning abilities in rats.
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Animals , Humans , Rats , Central Nervous System , Demyelinating Diseases , Encephalomyelitis, Autoimmune, Experimental , Excitatory Postsynaptic Potentials , Hippocampus , Learning Disabilities , Learning , Long-Term Potentiation , Memory , Memory Disorders , Models, Animal , Multiple Sclerosis , Neurons , Synaptic TransmissionABSTRACT
Objective To explore the effect of vasoactive intestinal peptide ( VIP) on the prevention and treatment of experimental autoimmune encephalomyelitis(EAE) rats by regulating the levels of IFN-γand IL-17A in brain tissue. Methods Sixty healthy female Wistar rats were randomly divided into 4 groups:normal control group, EAE control group, VIP low-dose group and VIP high-dose group.Myelin basic protein ( MBP)+complete Freurd′s adjuvant ( CFA) was used to establish an EAE model.The low and high-dose VIP groups were intraperitoneally injected with VIP 4 nmol/kg(0.2 ml) and 16 nmol/kg (0.8 ml) respectively every other day,while normal control group and EAE group with 0.8 ml saline for ten consecutive days.The incubation period, progression and peak of neurological dysfunction score ( NDS) changes of rats were recorded.The pathological changes, the GFAP+astrocyte activation in the brain at the morbidity peak of rats and the cytokine levels of IFN-γand IL-17A in brain tissue were observed.Results The incubation period was extended, the progression and peak NDS were shortened in the two VIP groups.In normal control group, there was no inflammatory cell infiltration or active astrocytes in brain tissue.The degree of infiltration of inflammatory cells and the degree of astrocyte activation in the VIP control group were significantly lower than in the EAE group.The cytokine levels of IFN-γand IL-17A in brain tissue were reduced in VIP groups.Conclusion By lowering IFN-γand IL-17A content in brain tissue, the infiltration of inflammatory cells and astrocyte activation are inhibited.VIP plays an important role in prevention and control of EAE.
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Objective To study the inhibition effect of non custodial terpenes-3β-alcohol to experimentally in-duced autoimmune encephalomyelitis in guinea pigs. Methods Different doses (25 mg/kg, 50 mg/kg and 100 mg/kg) of non custodial terpenes-3β-alcohol were given to the experimentally induced autoimmune encephalomyelitis model of guinea pigs by gavage for 8 weeks. Plasma levels of CD4+/CD8+, IL-1, IL-2, IL-6, IL-10, neuropeptide Y (NPY), beta endorphin (β-EP) , transforming growth factor-β(TGF-β), matrix metalloproteinase (MMP-2), nitric oxide synthase (NOS) and leuko-cyte differentiation antigen CD3 were assessed. The brain neuron morphology changes was observed under light microscopy while its ultrastructure changes was observed under electron microscope. NOS expression in neurons was observed through immunofluoresce technology. Results Non custodialterpenes-3β-alcohol inhibited the increase of plasma CD4+/CD8+, IL-1, IL-2, IL-6, IL-10, MMP-2, CD3 and NPY while decrease of plasmaβ-EP, brain TGF-β. It also increase NOS expres-sion in neuronal cytoplasm and maintained neuron morphology. Conclusion Non custodial terpenes-3β-alcohol inhibit-ed the experimental autoimmune encephalomyelitis in guinea pig.
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Objective To study the regulation effect of estrogen in expression of matrix metalloproteinase-9 (MMP-9) in the central nervous system (CNS) in mice with experimental autoimmune encephalomyelitis ( EAE).Methods The 60 mice were overiectomized and 2 weeks later EAE was induced with MOG35-55 peptide in these mice.They were divided into a treatment group and a control group.The treatment group was treated with estrogen and the control group was given PBS.Clinical symptoms in these two groups were scored and compared.HE staining was used to observe inflammation in the brain and spinal cord.The MMP-9 expression in the CNS was examined by quantitative real-time PCR and immunofluorescence staining.Results The incidence of disease was lower (treatment and control group were 8/30 and 28/30 respectively) and clinical symptoms were milder (treatment and control group were 3.23±0.83 and 1.62 ±1.00 respectively,t=3.811 and P＜0.05) in the treatment group than those in the control group.HE staining showed the decreased infiltration of inflammatory cell in the treatment group (Treatment group:inflammatory score were 0.895 ±0.206,0.752 ±0.302,0.732 ±0.183 in acute,relief and chronic phase respectively;Control group:inflammatory score were 3.472 ±0.635,2.881 ±0.662,1.891 ± 0.482 in acute,relief and chronic phase respectively.t = 8.622,6.543 and 5.027,all P ＜ 0.05).The quantitative real-time PCR and immunofluorescence staining showed that the expression of MMP9 in the CNS was decreased in the treatment group.Conclusion Estrogen may decrease MMP-9 expression in the CNS,reduce inflammation and clinical symptoms in mice with EAE.
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Objective To investigate the effect of ulinastatin (UTI) on the expression of brainderived neurotrophic factor ( BDNF ) and remyelination in mice with experimental autoimmune encephalomyelitis ( EAE).Methods Twenty-four C57BL/6 mice were randomly divided into UTI group (U),normal saline treated group (S) and normal control group (N,n = 8,respectively).Demyelinations in the spinal cord were observed by solochrome cyanin staining.The expression of BDNF,myelin basic protein (MBP),and 2',3 '-cyclic nucleotide 3'-phosphodiesterase (CNP) in brain tissue of each group were evaluated by Western blot.Results Average clinical scores in group U at the 12,13,14,22,23,31,33,34 and 35 days were 0,0.25,0.38,0.63,0.63,0.40,0.40,0.40 and 0.40 respectively.They were significantly lower than group S at the same time ( U= 16.00,15.00,14.50,7.50,0.00,14.50,14.50,12.00 and 14.50,all P ＜0.05).Solochrome cyanin staining showed that demyelination of spinal cord in group U was also significantly improved than group S.Expressions of BDNF ( 1.96 ± 0.29),MBP (2.67 ± 0.48 ) and CNP ( 1.75 ± 0.20) in group U were all significantly higher than group S ( There were 0.80 ± 0.15,1.36 ± 0.38 and 1.06 ± 0.18 respectively,all P ＜ 0.05).Conclusions UTI has protective effect on EAE.The possible mechanism is that it could promote remyelination,and protect oligodendrocytes and neurons in EAE model by increasing BDNF expression in brain.
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Objective To explore the repair mechanism of olfactory ensheathing cells (OECs)-neurotrophin-3 (NT-3) gene engineering cell on neuron myeline and axon of experimental allergic encephalomyelitis (EAE).Methods OECs-NT-3 gene engineering cell, constructed by ueurotrophin-3 transinfecting OECs inducted by retrovirus, was transplanted into lateral ventricle.The migration and distribution were observed and compared with control group and OECs transplantation group.Then myeline repair and axon regeneration were evaluated in the aspects of function score, morphological structure, SYN grey level Results (1) OECs-NT-3 could survive, diffuse, migrate with axons, spread in the focus diffusely on the 28th day after transplantation.(2) OECs-NT-3 survived and migrated to the transcription level of NT-3 mRNA in transgene group, being (212.3±16.1)×10-2, significantly higher than OECs group ((1.98±0.19)×10-2) and the contrast group ((1.23±0.13)×10-2, t = - 31.161, -31.928, P < 0.01).(3) The myeline of transgene group was kept complete and the number of inflamatory focus was lower than those of other groups (t = 11.388-22.728, P <0.01).(4) The SYN grey level of transgene group was obviously higher (P < 0.01).Conclusion OECs-NT-3 cell expresses NT-3 in EAE stably and effectively, which contributes to the repair of myeline and the regeneration of axon.
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A Esclerose Múltipla é uma doença crônica, inflamatória e desmielinizante do Sistema Nervoso Central. Embora a sua etiologia seja, ainda, desconhecida, supõe-se tratar-se de uma doença auto-imune mediada por células T CD4+ com perfil Th1. A Encefalomielite Auto-imune Experimental (EAE) é um modelo animal para estudar a Esclerose Múltipla. As características deste modelo são inflamações e desmielinização, se assemelhando à Esclerose Múltipla. A EAE permite avaliar parâmetros tais como linfócitos T CD4+ e T CD8+, linfócitos T regulatórios, moléculas co-estimulatórias, quimiocinas e etc. O objetivo deste estudo foi fazer uma revisão da literatura sobre imunopatologia da EAE murina mediada por linfócitos T.
Multiple Sclerosis is a chronic, inflammatory and demyelinating disease of the Central Nervous System. Although its etiology is still unclear, it is supposed a CD4+ T Helper-1- mediated autoimmune disease. Experimental Autoimmune Encephalomyelitis (EAE) is an animal model to study Multiple Sclerosis. The characteristics of this model are inflammation and demyelination similar to Multiple Sclerosis. EAE allows to assess parameters such as CD4+ and CD8+ T lymphocytes, regulatory T lymphocytes, costimulatory molecules, chemokines and so on. The aim of this study was to make a bibliographic revision of the murine EAE immunopathology mediated by T cells.
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Humans , Encephalomyelitis, Autoimmune, Experimental/immunology , Multiple Sclerosis/diagnosis , Autoimmune Diseases , T-Lymphocytes/immunologyABSTRACT
Intradermal gene administration was found to induce a more profound immune response than direct intramusclular gene injection. We performed intradermal vaccination of B10.PL mice with DNA encoding for the V 8.2 region of the T-cell receptors (TCR). Three weeks later, these mice were immunized with rat myelin basic protein (MBP). Daily mean clinical scores and mortality rate were lower in this group compared with controls. The proliferative responses of lymph node cells to rat MBP were slightly less in the vaccination groups than in the control groups (p<0.05). However, we detected no differences between the two groups with regard to the production of MBP-specific IgG, IgG1, & IgG2a antibodies. The levels of cytokine mRNA expression in the vaccination groups were observed higher than in the control groups without antigen-specific stimulation, but all of cytokine expressions between the vaccination and control groups after antigen-specific stimulation were identical. These results demonstrate that intradermal DNA vaccines encoding for TCR might prove to be useful in the control of autoimmune disease.