Multiple sclerosis is characterized by demyelination and neuronal loss caused by inflammatory cell activation and infiltration into the central nervous system.Macrophage polarization plays an important role in the pat...Multiple sclerosis is characterized by demyelination and neuronal loss caused by inflammatory cell activation and infiltration into the central nervous system.Macrophage polarization plays an important role in the pathogenesis of experimental autoimmune encephalomyelitis,a traditional experimental model of multiple sclerosis.This study investigated the effect of Fasudil on macrophages and examined the therapeutic potential of Fasudil-modified macrophages in experimental autoimmune encephalomyelitis.We found that Fasudil induced the conversion of macrophages from the pro-inflammatory M1 type to the anti-inflammatory M2 type,as shown by reduced expression of inducible nitric oxide synthase/nitric oxide,interleukin-12,and CD16/32 and increased expression of arginase-1,interleukin-10,CD14,and CD206,which was linked to inhibition of Rho kinase activity,decreased expression of toll-like receptors,nuclear factor-κB,and components of the mitogen-activated protein kinase signaling pathway,and generation of the pro-inflammatory cytokines tumor necrosis factor-α,interleukin-1β,and interleukin-6.Crucially,Fasudil-modified macrophages effectively decreased the impact of experimental autoimmune encephalomyelitis,resulting in later onset of disease,lower symptom scores,less weight loss,and reduced demyelination compared with unmodified macrophages.In addition,Fasudil-modified macrophages decreased interleukin-17 expression on CD4^(+)T cells and CD16/32,inducible nitric oxide synthase,and interleukin-12 expression on F4/80^(+)macrophages,as well as increasing interleukin-10 expression on CD4^(+)T cells and arginase-1,CD206,and interleukin-10 expression on F4/80^(+)macrophages,which improved immune regulation and reduced inflammation.These findings suggest that Fasudil-modified macrophages may help treat experimental autoimmune encephalomyelitis by inducing M2 macrophage polarization and inhibiting the inflammatory response,thereby providing new insight into cell immunotherapy for multiple sclerosis.展开更多
Emodin,a substance extracted from herbs such as rhubarb,has a protective effect on the central nervous system.However,the potential therapeutic effect of emodin in the context of multiple sclerosis remains unknown.In ...Emodin,a substance extracted from herbs such as rhubarb,has a protective effect on the central nervous system.However,the potential therapeutic effect of emodin in the context of multiple sclerosis remains unknown.In this study,a rat model of experimental autoimmune encephalomyelitis was established by immune induction to simulate multiple sclerosis,and the rats were intraperitoneally injected with emodin(20 mg/kg/d)from the day of immune induction until they were sacrificed.In this model,the nucleotide-binding domain-like receptor family pyrin domain containing 3(NLRP3)inflammasome and the microglia exacerbated neuroinflammation,playing an important role in the development of multiple sclerosis.In addition,silent information regulator of transcription 1(SIRT1)/peroxisome proliferator-activated receptor-alpha coactivator(PGC-1α)was found to inhibit activation of the NLRP3 inflammasome,and SIRT1 activation reduced disease severity in experimental autoimmune encephalomyelitis.Furthermore,treatment with emodin decreased body weight loss and neurobehavioral deficits,alleviated inflammatory cell infiltration and demyelination,reduced the expression of inflammatory cytokines,inhibited microglial aggregation and activation,decreased the levels of NLRP3 signaling pathway molecules,and increased the expression of SIRT1 and PGC-1α.These findings suggest that emodin improves the symptoms of experimental autoimmune encephalomyelitis,possibly through regulating the SIRT1/PGC-1α/NLRP3 signaling pathway and inhibiting microglial inflammation.These findings provide experimental evidence for treatment of multiple sclerosis with emodin,enlarging the scope of clinical application for emodin.展开更多
Multiple sclerosis is an autoimmune neurodegenerative disease of the central nervous system characterized by pronounced inflammatory infiltrates entering the brain,spinal cord and optic nerve leading to demyelination....Multiple sclerosis is an autoimmune neurodegenerative disease of the central nervous system characterized by pronounced inflammatory infiltrates entering the brain,spinal cord and optic nerve leading to demyelination.Focal demyelination is associated with relapsing-remitting multiple sclerosis,while progressive forms of the disease show axonal degeneration and neuronal loss.The tests currently used in the clinical diagnosis and management of multiple sclerosis have limitations due to specificity and sensitivity.MicroRNAs(miRNAs)are dysregulated in many diseases and disorders including demyelinating and neuroinflammatory diseases.A review of recent studies with the experimental autoimmune encephalomyelitis animal model(mostly female mice 6–12 weeks of age)has confirmed miRNAs as biomarkers of experimental autoimmune encephalomyelitis disease and importantly at the pre-onset(asymptomatic)stage when assessed in blood plasma and urine exosomes,and spinal cord tissue.The expression of certain miRNAs was also dysregulated at the onset and peak of disease in blood plasma and urine exosomes,brain and spinal cord tissue,and at the post-peak(chronic)stage of experimental autoimmune encephalomyelitis disease in spinal cord tissue.Therapies using miRNA mimics or inhibitors were found to delay the induction and alleviate the severity of experimental autoimmune encephalomyelitis disease.Interestingly,experimental autoimmune encephalomyelitis disease severity was reduced by overexpression of miR-146a,miR-23b,miR-497,miR-26a,and miR-20b,or by suppression of miR-182,miR-181c,miR-223,miR-155,and miR-873.Further studies are warranted on determining more fully miRNA profiles in blood plasma and urine exosomes of experimental autoimmune encephalomyelitis animals since they could serve as biomarkers of asymptomatic multiple sclerosis and disease course.Additionally,studies should be performed with male mice of a similar age,and with aged male and female mice.展开更多
Sinomenine is a bioactive alkaloid isolated from the Chinese medicinal plant Sinomenium acutum.It is widely used as an immunosuppressive drug for treating rheumatic and arthritic diseases.In our previous studies,we fo...Sinomenine is a bioactive alkaloid isolated from the Chinese medicinal plant Sinomenium acutum.It is widely used as an immunosuppressive drug for treating rheumatic and arthritic diseases.In our previous studies,we found that sinomenine reduced cellular infiltration within the spinal cord and alleviated experimental autoimmune encephalomyelitis(EAE) in rats.In this study,we further investigated the mechanisms of sinomenine treatment in EAE rats.In EAE rats,treatment with sinomenine exerted an anti-inducible NO synthase(anti-iNOS) effect,which is related to the reductions of Th1 cytokine interferon-γ(IFN-γ) and its transcription factor,T-bet,in spinal cords.Moreover,sinomenine treatment of splenocytes stimulated with anti-CD3 antibody and recombinant rat interleukin 12 reduced the expression of T-bet and IFN-γ in vitro and also reduced the capability of supernatants of splenocyte culture to induce iNOS expression by primary astrocytes.However,sinomenine had no direct inhibitory effect on iNOS produced by astrocytes cultured with IFN-γ and tumor necrosis factor α in vitro.In conclusion,the anti-iNOS effect of sinomenine on EAE is mediated via the suppression of T-bet /IFN-γ pathway.展开更多
BACKGROUND:Previous studies have focused on the correlation between Nogo-A expression and multiple sclerosis or between Nogo-A receptor(NgR) expression and multiple sclerosis in the central nervous system.Expression p...BACKGROUND:Previous studies have focused on the correlation between Nogo-A expression and multiple sclerosis or between Nogo-A receptor(NgR) expression and multiple sclerosis in the central nervous system.Expression patterns of Nogo-A and NgR remain poorly understood in rat models of experimental autoimmune encephalomyelitis(EAE).OBJECTIVE:To observe dynamic changes in Nogo-A and NgR protein expression,and to verify the correlation between Nogo-A and NgR protein,as well as expression patterns at various time points,in periventricular tissue of EAE rats.DESIGN,TIME AND SETTING:A neuroimmunological,randomized,controlled experiment was performed at the Clinical Institute of Hunan People's Hospital of China from September to November 2008.MATERIALS:Immunohistochemistry(streptavidin-biotin-peroxidase complex method) kit was purchased from Boster,China.METHODS:A total of 60 female,Wistar rats,aged 6-8 weeks,were randomly assigned to EAE and control groups(n = 30,respectively).Guinea pig spinal cord homogenate,self-made complete Freund's adjuvant(0.2 mL/100 g),and pertussis vaccine(0.2 mL) were subcutaneously injected into the hindlimb foot pad of rats from the EAE group to create rat models of EAE.Complete Freund's adjuvant(0.2 mL) was infused into rats from the control group.MAIN OUTCOME MEASURES:Nogo-A and NgR protein expression was determined in periventricular white matter using immunohistochemical methods.Neurological scores were determined in all rats.RESULTS:Rats from the EAE group developed acute-onset EAE following immunization.The pathogenetic symptoms reached a peak on day 15,and neurological scores were also greatest at this time point.Neurological scores decreased with recovery of the illness.Nogo-A was shown to be expressed in neuronal cells and oligodendrocytes,and expression increased 11 days after immunization(P < 0.01),decreased by day 13(P < 0.01),and then increased again by day 15.Nogo-A expression remained greater in the EAE group compared with the control group at day 30(P < 0.01).In the EAE group,NgR protein was primarily expressed on the surface of neuronal bodies and axons.NgR expression increased 13-18 days after immunization(P < 0.01 or P < 0.05).CONCLUSION:Nogo-A and NgR protein expression altered with disease course in periventricular white matter of EAE rats.Results suggested that Nogo-A and NgR were involved in EAE occurrence.展开更多
OBJECTIVE Experimental autoimmune encephalomyelitis(EAE),the classical animal model for multiple sclerosis(MS)is triggered by an impaired balance of T helper(Th)cells and regulatory T(Tregs)cells.Matrine(MAT),a quinol...OBJECTIVE Experimental autoimmune encephalomyelitis(EAE),the classical animal model for multiple sclerosis(MS)is triggered by an impaired balance of T helper(Th)cells and regulatory T(Tregs)cells.Matrine(MAT),a quinolizidine alkaloid derived from the herb Radix Sophorae Flave,has been shown to ameliorate the clinical signs,inflammatory infiltration,demyelination in acute EAE rats.However,whether MAT protect from EAE by adjusting Th and Treg cells response in specific-cellular and molecular level is unknown.METHODS Herein,MAT was tested for its effects on Th1,Th2,Th17 and Treg cells in the spinal cord of EAE mice and splenocyte-extracted from EAE mice with MOG35-55-restimulated,respectively.RESULTS Our findings revealed that MAT significantly inhibit the proliferation of splenocyte,and remarkably down-regulate the differentiation of Th1/Th17 cells with decreased expressions of CD4+IFN-γ+cells and CD4+IL-17+cells in vivo and IL-17,IFN-γ,ROR-γt,T-bet in vitro,meanwhile it dramatically up-regulate the Th2/Treg cells response associated with increased levels of CD4+TGF-β+1cells and CD4+IL-10+cells in vivo and IL-4,IL-10,TGF-β1,Foxp3 and GATA3in vitro.CONCLUSION Considering the effective therapeutic effects of MAT on EAE,it′s worth to find its new values on other autoimmune diseases.展开更多
AIM: To investigate the temporal onset and dynamic interplay of CD4+ T helper cell subsets in experimental autoimmune encephalomyelitis(EAE).METHODS: EAE was induced in C57BL/6 mice by immunization with myelin oligode...AIM: To investigate the temporal onset and dynamic interplay of CD4+ T helper cell subsets in experimental autoimmune encephalomyelitis(EAE).METHODS: EAE was induced in C57BL/6 mice by immunization with myelin oligodendrocyte glycoprotein peptide p35-55. The clinical signs were scored and the tissue samples and immune cells isolated for analysis at different phases of EAE. The expression levels of inflammatory cytokines and related transcription factors were detected by quantitative reverse transcription polymerase chain reaction(PCR) and enzyme linked immunosorbant assay(ELISA). The percentages of Th1, Th17, Th2, Treg and memory T cell subsets in EAE were analyzed by immunostaining and flow cytometry.The data were analyzed by statistical techniques.RESULTS: Quantitative real-time PCR analysis showedthat EAE mice express elevated levels of Th1 [interferon gamma(IFNγ), interleukin(IL)-12p40 ], Th17 [IL-17, related orphan receptor gamma(RORγ), IL-12p40] and Treg [Foxp3, Epstein-Barr virus induced gene 3(EBI3), IL-10 ] genes in the central nervous system at the peak of the disease. Whereas, the expression of Th1(IFNγ, T-bet, IL-12p35, IL-12p40), Th17(RORγ, IL-12p40), Th2(IL-4) and Treg(Foxp3, EBI3) response genes was reduced in the spleen during pre-disease but gradually recovered at the later phases of EAE. ELISA and flow cytometry analyses showed an increase in Th17 response in the periphery, while Th1 response remained unchanged at the peak of disease. The m RNA levels of IFNγ, IL-17 and IL-12p40 in the brain were increased by 23(P < 0.001), 9(P < 0.05) and 14(P < 0.01) fold, respectively, on day 21 of EAE. Conversely, the mR NA expression of IL-10 was increased by 2 fold(P < 0.05) in the spleen on day 21. CD4+CD25+Foxp3+Treg response was reduced at pre-disease but recovered to naíve levels by disease onset. The percentage of CD25+Foxp3+ regulatory T cells decreased from 7.7% in the naíve to 3.2%(P < 0.05) on day 7 of EAE, which then increased to 8.4% by day 28. Moreover, the CD4+CD127+CD44high memory T cell response was increased during the onset and recovery phases of EAE. The memory and effector cells showed an inverse relationship in EAE, where the memory T cells increased from 12.3% in naive to 20% by day 21, and the effector cells decreased from 32% in naíve to 21%(P < 0.01) by day 21. The wild type C57BL/6 mice with EAE showed elevated levels of effector-memory T cells(TEM) with concomitant reduction in central-memory T cells(TCM), but the EAE-resistant IL-7R deficient mice showed elevated TCM with no effect on TEM cells in EAE.CONCLUSION: Our findings highlight the temporal onset and dynamic interplay of effector, memory and regulatory CD4+ T cell subsets and its significance to clinical outcome in EAE and other autoimmune diseases.展开更多
The transcription factor nuclear factor κB(NF-κB) plays major roles in inflammatory diseases through regulation of inflammation and cell viability.Multiple sclerosis(MS) is a chronic inflammatory demyelinating and n...The transcription factor nuclear factor κB(NF-κB) plays major roles in inflammatory diseases through regulation of inflammation and cell viability.Multiple sclerosis(MS) is a chronic inflammatory demyelinating and neurodegenerative disease of the central nervous system(CNS).It has been shown that NF-κB is activated in multiple cell types in the CNS of MS patients,including T cells,microglia/macrophages,astrocytes,oligodendrocytes,and neurons.Interestingly,data from animal model studies,particularly studies of experimental autoimmune encephalomyelitis,have suggested that NF-κB activation in these individual cell types has distinct effects on the development of MS.In this review,we will cover the current literature on NF-κB and the evidence for its role in the development of MS and its animal model experimental autoimmune encephalomyelitis.展开更多
<正>BACKGROUND:Yishendaluo decoction reduces production of inflammatory mediators,relieves damage due to inflammatory reactions,and improves neural functions during experimental autoimmune encephalomyelitis. OBJ...<正>BACKGROUND:Yishendaluo decoction reduces production of inflammatory mediators,relieves damage due to inflammatory reactions,and improves neural functions during experimental autoimmune encephalomyelitis. OBJECTIVE:To investigate the effects of Yishendaluo decoction on a mouse model of experimental autoimmune encephalomyelitis. DESIGN,TIME AND SETTING:The randomized,controlled,neuropathological,and molecular biological animal study was performed at the Key Laboratory of Chinese Internal Medicine,Ministry of Education,Dongzhimen Hospital of Beijing University of Chinese Medicine and Center for Neuroinformatics,General Hospital of Chinese PLA from 2005 to 2006. MATERIALS:Yishendaluo decoction pieces consisting of prepared rehmannia root,colla cornus cervi,cape jasmine fruit,and grassleaf sweetflag rhizome were purchased from the Dongzhimen Hospital of Beijing University of Chinese Medicine.Rabbit anti-mouseβ-amyloid precursor protein and p38 polyclonal antibody(Zhongshan Goldenbridge Biotechnology,China),as well interferon-γand interleukin-4 ELISA kit(Boster,China),were used in this study. METHODS:A total of 96 healthy,female,SJL/J mice,aged 8-12 weeks,were equally and randomly assigned to normal,model,hormone,and Chinese medicine groups.A total of 0.2 mL antigen preparation,supplemented with 150μg PLP139-151 and 400μg H37RA,was subcutaneously injected into the upper abdomen of mice from the model,hormone,and Chinese medicine groups. Mouse models of experimental autoimmune encephalomyelitis were established by intravenous injection of 0.1 mL Bordetella pertussis solution containing 0.6×10~6 Bordetella pertussis at days 1 and 3.Mice from the model,Chinese medicine,and hormone groups were respectively subjected to 0.2 mL saline,2 g/kg Yishendaluo decoction,and 0.078 mg/kg prednisone acetate,once daily for 14 consecutive days.Mice from the normal group were left intact. MAIN OUTCOME MEASURES:Pathological changes were observed using hematoxylin-eosin staining and Luxol fast blue staining.Expression ofβ-amyloid precursor protein and p38 protein was determined by immunohistochemistry.Levels of interferon-γand interleukin-4 were detected by ELISA.Behavioral changes were assessed in mice according to scores of neurological function. RESULTS:A few inflammatory cell infiltration,nerve fiber breakage and slight demyelination were detected in the central nervous system of mice from the Chinese medicine and hormone groups compared with the model group.Expression ofβ-amyloid precursor protein and p38 protein was significantly diminished in the central nervous system of mice from the Chinese medicine and hormone groups compared with the model group(P<0.05 or P<0.01),and the decrease was greatest in the Chinese medicine group.The decrease in mouse weight was not significant,and neurological function scores were less in the Chinese medicine and hormone groups compared with the model group(P<0.05 or P<0.01).Interferon-γlevels were significantly reduced(P<0.01),and interleukin-4 levels were significantly increased(P<0.01) in the brains of the Chinese medicine and hormone groups,compared with the model group. CONCLUSION:Yishendaluo decoction improved neurological function in mice with experimental autoimmune encephalomyelitis by downregulatingβ-amyloid precursor protein expression,resisting axonal degeneration,and relieving inflammatory reaction.The anti-inflammatory mechanism was regulated by inhibition of the p38 mitogen-activated protein kinase signal pathway.展开更多
Studies have demonstrated that amyloid precursor protein(APP)expression increases in multiple sclerosis tissues during acutely and chronically active stages.To determine the relationship between axonal injury and rege...Studies have demonstrated that amyloid precursor protein(APP)expression increases in multiple sclerosis tissues during acutely and chronically active stages.To determine the relationship between axonal injury and regeneration in multiple sclerosis,an animal model of experimental autoimmune encephalomyelitis was induced using different doses of myelin basic protein peptide.APP and growth-associated protein 43(GAP-43),which is considered a specific marker of neural regeneration,were assessed by western blot analysis.Expression of APP and GAP-43,as well as the correlation between these two proteins,in brain white matter and spinal cord tissues of experimental autoimmune encephalomyelitis rats at different pathological stages was analyzed.Results showed that APP and GAP-43 expression increased during the acute stage and decreased during remission,with a positive correlation between APP and GAP-43 expression in brain white matter and spinal cord tissues.These results suggest that APP and GAP-43 could provide nutritional and protective effects on damaged neurons.展开更多
Multiple sclerosis(MS) is a common demyelinating central nervous system disease associated with progressive physical impairment. To study the mechanism underlying disease pathogenesis and develop potential treatments,...Multiple sclerosis(MS) is a common demyelinating central nervous system disease associated with progressive physical impairment. To study the mechanism underlying disease pathogenesis and develop potential treatments, experimental autoimmune encephalomyelitis(EAE) is often used as an animal model. EAE can be induced in various species by introducing specific antigens, which ultimately result in motor dysfunction. Although the severity of the paralysis is indicated using the EAE score, there is no standard scoring system for EAE signs, and there is variability between research groups with regard to the exact EAE scoring system utilized. Here, we describe the criteria used for EAE scoring systems in various laboratories and suggest combining EAE score with another quantitative index to evaluate paralysis, such as the traveled distance, with the goal of facilitating the study of the mechanisms and treatment of MS.展开更多
This study investigated the effects of Yishendaluo decoction on the loss of blood-brain barrier integrity in mice exhibiting experimental autoimmune encephalomyelitis.To this end,we used real-time fluorescent quantita...This study investigated the effects of Yishendaluo decoction on the loss of blood-brain barrier integrity in mice exhibiting experimental autoimmune encephalomyelitis.To this end,we used real-time fluorescent quantitative PCR to measure the levels of mRNAs specific to the T cell markers CD4 and CD8,and the monocyte marker CD11b.In addition,we used Evans blue dye extravasation in the spinal cord and brain tissues to assess blood-brain barrier permeability.The results indicated that an increase in blood-brain barrier permeability was associated with an increase in CD4,CD8 and CD11b mRNA expression in experimental autoimmune encephalomyelitis mice.Yishendaluo decoction administration significantly reversed inflammatory cell accumulation in cerebral tissues of experimental autoimmune encephalomyelitis mice.展开更多
Myelin-associated inhibitory factors within the central nervous system(CNS) are considered to be one of the main obstacles for axonal regeneration following disease or injury. The nogo receptor 1(NgR1) has been well d...Myelin-associated inhibitory factors within the central nervous system(CNS) are considered to be one of the main obstacles for axonal regeneration following disease or injury. The nogo receptor 1(NgR1) has been well documented to play a key role in limiting axonal regrowth in the injured and diseased mammalian CNS. However, the role of nogo receptor in immune cell activation during CNS inflammation is yet to be mechanistically elucidated. Microglia/macrophages are immune cells that are regarded as pathogenic contributors to inflammatory demyelinating lesions in multiple sclerosis(MS). In this study, the animal model of MS, experimental autoimmune encephalomyelitis(EAE) was induced in ngr1^(+/+) and ngr1^(–/–) female mice following injection with the myelin oligodendrocyte glycoprotein(MOG_(35–55)) peptide. A fatemap analysis of microglia/macrophages was performed throughout spinal cord sections of EAE-induced mice at clinical scores of 0, 1, 2 and 3, respectively(increasing locomotor disability) from both genotypes, using the CD11 b and Iba1 cell markers. Western immunoblotting using lysates from isolated spinal cord microglia/macrophages, along with immunohistochemistry and flow cytometric analysis, was performed to demonstrate the expression of nogo receptor and its two homologs during EAE progression. Myelin protein engulfment during EAE progression in ngr1^(+/+) and ngr1^(–/–) mice was demonstrated by western immunblotting of lysates from isolated spinal cord microglia/macrophages, detecting levels of Nogo-A and MOG. The numbers of M1 and M2 microglia/macrophage phenotypes present in the spinal cords of EAE-induced ngr1^(+/+) and ngr1^(–/–) mice, were assessed by flow cytometric analysis using CD38 and Erg-2 markers. A significant difference in microglia/macrophage numbers between ngr1^(+/+) and ngr1^(–/–) mice was identified during the progression of the clinical symptoms of EAE, in the white versus gray matter regions of the spinal cord. This difference was unrelated to the expression of Ng R on these macrophage/microglial cells. We have identified that as EAE progresses, the phagocytic activity of microglia/macrophages with myelin debris, in ngr1^(–/–) mice, was enhanced. Moreover, we show a modulation from a predominant M1-pathogenic to the M2-neurotrophic cell phenotype in the ngr1^(–/–) mice during EAE progression. These findings suggest that CNS-specific macrophages and microglia of ngr1^(–/–) mice may exhibit an enhanced capacity to clear inhibitory molecules that are sequestered in inflammatory lesions.展开更多
Zuogui pills have been shown to attenuate the inflammatory reaction in a rat model of experimental autoimmune encephalomyelitis(EAE).The present study attempted to investigate the pathology underlying the influence of...Zuogui pills have been shown to attenuate the inflammatory reaction in a rat model of experimental autoimmune encephalomyelitis(EAE).The present study attempted to investigate the pathology underlying the influence of Zuogui pills on myelinolysis in EAE rats.Hematoxylin-eosin and Luxol fast blue staining showed that the myelinolysis foci in the cerebrum,cerebellum,brain stem,and the spinal cord of EAE rats were significantly decreased,along with serum myelin basic protein content following treatment with Zuogui pills.展开更多
The present study established a chronic experimental autoimmune encephalomyelitis model in C57BL/6 mice induced by myelin oligodendrocyte glycoprotein peptides and complete Freund's adjuvant.Onset latency was 12 d...The present study established a chronic experimental autoimmune encephalomyelitis model in C57BL/6 mice induced by myelin oligodendrocyte glycoprotein peptides and complete Freund's adjuvant.Onset latency was 12 days,with an incidence rate of 100%.Neuropathological characteristics included perivascular inflammatory cell infiltration,demyelination,neuronal degeneration,and axonal damage within cerebral and myelic white matter.Electron microscopy revealed swollen mitochondria,complete organ disappearance,and fused or broken myelin sheath structure,which were accompanied by myelin sheath reconstruction.Moreover,axonal damage was not consistent with demyelination distribution,and severity of axonal damage did not correlate with demyelination.Results suggested that axonal damage in an experimental autoimmune encephalomyelitis model is not secondary to inflammatory demyelination.展开更多
OBJECTIVE In order to investigate whether cornel iridoid glycoside(CIG),the main component extracted from Cornus officinalis,can treat demyelinating diseases of the central nervous system(CNS)such as multiple sclerosi...OBJECTIVE In order to investigate whether cornel iridoid glycoside(CIG),the main component extracted from Cornus officinalis,can treat demyelinating diseases of the central nervous system(CNS)such as multiple sclerosis(MS).METHODS CIG(30,60 and 120mg·kg-1)or vehicle was intragastrically administered once daily to rats,starting immediately after purified myelin basic protein(MBP)68-86 peptides immunization until day 20 post immunization(p.i.).Histopathological staining,enzyme-linked immunosorbent assay,biochemical methods and Western blotting approaches were used to evaluate the disease incidence and severity,neuroinflammatory and neurotrophic response in the CNS.RESULTS Neurological deficit and proportion of incidence seen in EAE rats were significantly reduced by CIG treatment in a dose-dependent manner.Histopathological staining showed that CIG treatment alleviated demyelination and inflammatory infiltration,increased the number of oligodendrocytes,enhanced the expression of brain-derived neurotrophic factor(BDNF).Production of proinflammatory molecules such as interleukin-1β(IL-1β),tumour necrosis factor-αand interferon-γwere also inhibited by CIG administration.CIG could ameliorate phosphorylation of STAT1,STAT3 and JAK1 as well as IL-6/IL-6 Rexpression,which involved in immune response and inflammation.CONCLUSION Our results demonstrated that CIG may ameliorate EAE rats through down-regulation of JAK/STAT signaling pathway.This study gave new insight into the novel regulatory mechanism of CIG and highlight novel therapeutic targets and a potential therapeutic agent for the treatment of MS.展开更多
Therapeutic administration of fingolimod hydrochloride (FTY720), the functional antagonist at sphingosine 1-phosphate (S1P) receptor 1 (S1P1) shows a marked improving effect on experimental autoimmune encephalomyeliti...Therapeutic administration of fingolimod hydrochloride (FTY720), the functional antagonist at sphingosine 1-phosphate (S1P) receptor 1 (S1P1) shows a marked improving effect on experimental autoimmune encephalomyelitis (EAE) induced by myelin oligodendrocyte glycoprotein (MOG) in C57BL/6 mice. However, this treatment showed an only partial inhibition of Th1/Th17 cell infiltration into the central nervous system (CNS), suggesting that down-regulation of lymphocytic S1P1 is insufficient to explain the therapeutic effect of FTY720 on EAE. On the other hand, the therapeutic administration of FTY720 reduced the mRNA expressions of IL-6, CCL2, and glial fibrillary acidic protein, an activation marker of astrocytes, in the CNS of EAE mice. In human astrocytic glyoma, U373MG cells, mRNA expression of S1P1 was higher as compared with those of the other S1P receptor subtypes and phosphorylation of Akt was induced by S1P, FTY720-phosphate (FTY720-P), or an S1P1-selective agonist, SEW2871. FTY720-P appeared to induce down-regulation of S1P1 in U373MG cells, implying a functional antagonism at S1P1 on astrocytes. S1P but not FTY720-P induced production of IL-6, IL-8, and CCL2 significantly and treatment with FTY720-P or SEW2871 inhibited production of these pro-inflammatory cytokines from U373MG cells stimulated with S1P. These results suggest that S1P-S1P1 axis induces production of pro-inflammatory cytokines by astrocytes. Consequently, it is highly probable that the therapeutic effects of FTY720 on EAE are caused by inhibiting not only egress of myelin-specific Th cells from the draining lymph nodes but also activation of astrocytes in the CNS.展开更多
Infiltration of myelin-specific helper T (Th) cells into the central nervous system (CNS) plays a key role in pathogenesis of experimental autoimmune encephalomyelitis (EAE). In this study, we investigated the involve...Infiltration of myelin-specific helper T (Th) cells into the central nervous system (CNS) plays a key role in pathogenesis of experimental autoimmune encephalomyelitis (EAE). In this study, we investigated the involvement of sphingosine 1-phosphate (S1P)-S1P receptor 1 (S1P1) axis in lymphocytes for EAE development when C57BL/6 mice were immunized with myelin oliogodendrocyte glycoprotein (MOG). The expression of S1P1 mRNA and S1P responsiveness of lymphocytes in draining lymph nodes (DLN) were down-regulated markedly after MOG immunization until onset of EAE. Accompanying with reacquisition of down-regulated S1P1 transcript and S1P responsiveness in DLN lymphocytes, MOG-immunized mice developed EAE symptoms with significant infiltration of Th1 and Th17 cells into the CNS and a marked elevation of IFN-γ, T-bet, IL-17, and RORγt mRNA expressions. Prophylactic administration of an S1P1 functional antagonist, fingolimod hydrochloride (FTY720, 0.3 mg/kg, orally) significantly inhibited EAE development and almost completely prevented infiltration of Th1 and Th17 cells into the CNS with a marked reduction of IFN-γ, T-bet, IL-17, and RORγt mRNA expressions. Similar results were obtained by treatment with an S1P1-selective agonist, SEW2871 or an S1P lyase inhibitor, 2-acetyl-4-tetrahydroxybutylimidazole. Moreover, FTY720-phosphate and SEW2871 inhibited in vitro migration of Th1 and Th17 cells toward S1P but did not affect cytokine production or generation of Th1 or Th17 cells. These results suggest that reacquisition of S1P1 expression in DLN lymphocytes plays a major role in trafficking of myelin antigen-specific Th1/Th17 cells from DLN to the CNS in EAE and that prophylactic effect of FTY720 on EAE is predominantly caused by functional antagonism via lymphocytic S1P1.展开更多
AIM: To study the effect of blocking the eo-2 pathwayon the development and severity of experimental autoimmune encephalomyelitis(EAE). METHODS: We produced m Ab directed against eo-2,named D8. MOG35-55 induced-EAE mi...AIM: To study the effect of blocking the eo-2 pathwayon the development and severity of experimental autoimmune encephalomyelitis(EAE). METHODS: We produced m Ab directed against eo-2,named D8. MOG35-55 induced-EAE mice were dailyintravenously injected with either 25 μg or 100 μg D8,or with vehicle control alone [phosphate-buffered saline(PBS)], starting from day 0 post immunization and weremonitored for EAE clinical score(n = 10 in each group).Mice were sacrificed on day 58 and their sera were assessed for the presence of anti-myelin oligodendrocyteglycoprotein(anti-MOG) antibodies autoantibodies, aswell as for the profile of pro-inflammatory cytokines andchemokines. Histological analysis of brain sections wasperformed by hematoxylin and eosin staining.RESULTS: Daily treatment of EAE induced mice with D8 significantly decreased the severity of EAE symptoms. Treatment with both concentrations of D8 ameliorated EAE symptoms compared to PBS treated mice, starting from day 42 post immunization(0.89 ± 0.35 in D8 25 μg and D8 100 μg treated groups vs 2.11 ± 0.38 in the PBS treated group, P = 0.03). A significant improvement in EAE clinical score compared to total Ig G treated mice was observed with the higher concentration of D8(0.81 ± 0.38 in D8 100 μg treated group vs 2.11 ± 0.31 in Ig G1 treated group, on day 56 post immunization, P = 0.04). D8 treated mice with EAE did not significantly exhibit lower sera levels of anti-MOG autoantibodies compared to Ig G-treated mice. However, they expressed lower sera levels of the pro-inflammatory cytokines: tumor necrosis factor(7.8 ± 0.2 pg/m L in D8 100 μg treated mice vs 19.9 ± 3.4 pg/m L in Ig G treated mice, P = 0.005) and interferon-gamma(1.4 ± 0.6 pg/m L in D8 100 μg treated mice vs 3.6 ± 0.4 pg/m L in Ig G treated mice, P = 0.02), as well as reduced levels of the chemokine macrophage chemoattractant protein-1(27.2 ± 3.1 pg/m L in D8 100 μg treated mice vs 63.7 ± 12.3 pg/m L in Ig G treated mice, P = 0.03). These findings indicate that blocking the eo-2 pathway in EAE may affect not only eosinophil infiltration into the central nervous system(CNS), but also have an effect on monocytes and T cells, but not humoral, mediated responses. Histological analysis of the brains of D8 treated mice with EAE support that this treatment decreases immune cells infiltrates in the CNS.CONCLUSION: Taken together, these findings suggest a role for eo-2 in EAE pathogenesis and consequentially may support a therapeutic potential of anti-eo-2 neutralizing m Ab in multiple sclerosis.展开更多
基金supported by a grant from the Department of Science and Technology of Shanxi Province,China,No.20210302123477(to CL)Datong Bureau of Science and Technology of China,No.2020152(to CL)the Opening Foundation of Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine,No.2022-KF-03(to CL).
文摘Multiple sclerosis is characterized by demyelination and neuronal loss caused by inflammatory cell activation and infiltration into the central nervous system.Macrophage polarization plays an important role in the pathogenesis of experimental autoimmune encephalomyelitis,a traditional experimental model of multiple sclerosis.This study investigated the effect of Fasudil on macrophages and examined the therapeutic potential of Fasudil-modified macrophages in experimental autoimmune encephalomyelitis.We found that Fasudil induced the conversion of macrophages from the pro-inflammatory M1 type to the anti-inflammatory M2 type,as shown by reduced expression of inducible nitric oxide synthase/nitric oxide,interleukin-12,and CD16/32 and increased expression of arginase-1,interleukin-10,CD14,and CD206,which was linked to inhibition of Rho kinase activity,decreased expression of toll-like receptors,nuclear factor-κB,and components of the mitogen-activated protein kinase signaling pathway,and generation of the pro-inflammatory cytokines tumor necrosis factor-α,interleukin-1β,and interleukin-6.Crucially,Fasudil-modified macrophages effectively decreased the impact of experimental autoimmune encephalomyelitis,resulting in later onset of disease,lower symptom scores,less weight loss,and reduced demyelination compared with unmodified macrophages.In addition,Fasudil-modified macrophages decreased interleukin-17 expression on CD4^(+)T cells and CD16/32,inducible nitric oxide synthase,and interleukin-12 expression on F4/80^(+)macrophages,as well as increasing interleukin-10 expression on CD4^(+)T cells and arginase-1,CD206,and interleukin-10 expression on F4/80^(+)macrophages,which improved immune regulation and reduced inflammation.These findings suggest that Fasudil-modified macrophages may help treat experimental autoimmune encephalomyelitis by inducing M2 macrophage polarization and inhibiting the inflammatory response,thereby providing new insight into cell immunotherapy for multiple sclerosis.
基金supported by the National Natural Science Foundation of China,No.81771271Key Research and Development Program of Liaoning Province,No.2020JH2/10300047Outstanding Scientific Fund of Shengjing Hospital(all to JF).
文摘Emodin,a substance extracted from herbs such as rhubarb,has a protective effect on the central nervous system.However,the potential therapeutic effect of emodin in the context of multiple sclerosis remains unknown.In this study,a rat model of experimental autoimmune encephalomyelitis was established by immune induction to simulate multiple sclerosis,and the rats were intraperitoneally injected with emodin(20 mg/kg/d)from the day of immune induction until they were sacrificed.In this model,the nucleotide-binding domain-like receptor family pyrin domain containing 3(NLRP3)inflammasome and the microglia exacerbated neuroinflammation,playing an important role in the development of multiple sclerosis.In addition,silent information regulator of transcription 1(SIRT1)/peroxisome proliferator-activated receptor-alpha coactivator(PGC-1α)was found to inhibit activation of the NLRP3 inflammasome,and SIRT1 activation reduced disease severity in experimental autoimmune encephalomyelitis.Furthermore,treatment with emodin decreased body weight loss and neurobehavioral deficits,alleviated inflammatory cell infiltration and demyelination,reduced the expression of inflammatory cytokines,inhibited microglial aggregation and activation,decreased the levels of NLRP3 signaling pathway molecules,and increased the expression of SIRT1 and PGC-1α.These findings suggest that emodin improves the symptoms of experimental autoimmune encephalomyelitis,possibly through regulating the SIRT1/PGC-1α/NLRP3 signaling pathway and inhibiting microglial inflammation.These findings provide experimental evidence for treatment of multiple sclerosis with emodin,enlarging the scope of clinical application for emodin.
文摘Multiple sclerosis is an autoimmune neurodegenerative disease of the central nervous system characterized by pronounced inflammatory infiltrates entering the brain,spinal cord and optic nerve leading to demyelination.Focal demyelination is associated with relapsing-remitting multiple sclerosis,while progressive forms of the disease show axonal degeneration and neuronal loss.The tests currently used in the clinical diagnosis and management of multiple sclerosis have limitations due to specificity and sensitivity.MicroRNAs(miRNAs)are dysregulated in many diseases and disorders including demyelinating and neuroinflammatory diseases.A review of recent studies with the experimental autoimmune encephalomyelitis animal model(mostly female mice 6–12 weeks of age)has confirmed miRNAs as biomarkers of experimental autoimmune encephalomyelitis disease and importantly at the pre-onset(asymptomatic)stage when assessed in blood plasma and urine exosomes,and spinal cord tissue.The expression of certain miRNAs was also dysregulated at the onset and peak of disease in blood plasma and urine exosomes,brain and spinal cord tissue,and at the post-peak(chronic)stage of experimental autoimmune encephalomyelitis disease in spinal cord tissue.Therapies using miRNA mimics or inhibitors were found to delay the induction and alleviate the severity of experimental autoimmune encephalomyelitis disease.Interestingly,experimental autoimmune encephalomyelitis disease severity was reduced by overexpression of miR-146a,miR-23b,miR-497,miR-26a,and miR-20b,or by suppression of miR-182,miR-181c,miR-223,miR-155,and miR-873.Further studies are warranted on determining more fully miRNA profiles in blood plasma and urine exosomes of experimental autoimmune encephalomyelitis animals since they could serve as biomarkers of asymptomatic multiple sclerosis and disease course.Additionally,studies should be performed with male mice of a similar age,and with aged male and female mice.
基金supported by Science Fund of the Health Department of Jiangsu Province (No. H200504)
文摘Sinomenine is a bioactive alkaloid isolated from the Chinese medicinal plant Sinomenium acutum.It is widely used as an immunosuppressive drug for treating rheumatic and arthritic diseases.In our previous studies,we found that sinomenine reduced cellular infiltration within the spinal cord and alleviated experimental autoimmune encephalomyelitis(EAE) in rats.In this study,we further investigated the mechanisms of sinomenine treatment in EAE rats.In EAE rats,treatment with sinomenine exerted an anti-inducible NO synthase(anti-iNOS) effect,which is related to the reductions of Th1 cytokine interferon-γ(IFN-γ) and its transcription factor,T-bet,in spinal cords.Moreover,sinomenine treatment of splenocytes stimulated with anti-CD3 antibody and recombinant rat interleukin 12 reduced the expression of T-bet and IFN-γ in vitro and also reduced the capability of supernatants of splenocyte culture to induce iNOS expression by primary astrocytes.However,sinomenine had no direct inhibitory effect on iNOS produced by astrocytes cultured with IFN-γ and tumor necrosis factor α in vitro.In conclusion,the anti-iNOS effect of sinomenine on EAE is mediated via the suppression of T-bet /IFN-γ pathway.
文摘BACKGROUND:Previous studies have focused on the correlation between Nogo-A expression and multiple sclerosis or between Nogo-A receptor(NgR) expression and multiple sclerosis in the central nervous system.Expression patterns of Nogo-A and NgR remain poorly understood in rat models of experimental autoimmune encephalomyelitis(EAE).OBJECTIVE:To observe dynamic changes in Nogo-A and NgR protein expression,and to verify the correlation between Nogo-A and NgR protein,as well as expression patterns at various time points,in periventricular tissue of EAE rats.DESIGN,TIME AND SETTING:A neuroimmunological,randomized,controlled experiment was performed at the Clinical Institute of Hunan People's Hospital of China from September to November 2008.MATERIALS:Immunohistochemistry(streptavidin-biotin-peroxidase complex method) kit was purchased from Boster,China.METHODS:A total of 60 female,Wistar rats,aged 6-8 weeks,were randomly assigned to EAE and control groups(n = 30,respectively).Guinea pig spinal cord homogenate,self-made complete Freund's adjuvant(0.2 mL/100 g),and pertussis vaccine(0.2 mL) were subcutaneously injected into the hindlimb foot pad of rats from the EAE group to create rat models of EAE.Complete Freund's adjuvant(0.2 mL) was infused into rats from the control group.MAIN OUTCOME MEASURES:Nogo-A and NgR protein expression was determined in periventricular white matter using immunohistochemical methods.Neurological scores were determined in all rats.RESULTS:Rats from the EAE group developed acute-onset EAE following immunization.The pathogenetic symptoms reached a peak on day 15,and neurological scores were also greatest at this time point.Neurological scores decreased with recovery of the illness.Nogo-A was shown to be expressed in neuronal cells and oligodendrocytes,and expression increased 11 days after immunization(P < 0.01),decreased by day 13(P < 0.01),and then increased again by day 15.Nogo-A expression remained greater in the EAE group compared with the control group at day 30(P < 0.01).In the EAE group,NgR protein was primarily expressed on the surface of neuronal bodies and axons.NgR expression increased 13-18 days after immunization(P < 0.01 or P < 0.05).CONCLUSION:Nogo-A and NgR protein expression altered with disease course in periventricular white matter of EAE rats.Results suggested that Nogo-A and NgR were involved in EAE occurrence.
基金The project supported by National Natural Science Foundation of China(31570357)
文摘OBJECTIVE Experimental autoimmune encephalomyelitis(EAE),the classical animal model for multiple sclerosis(MS)is triggered by an impaired balance of T helper(Th)cells and regulatory T(Tregs)cells.Matrine(MAT),a quinolizidine alkaloid derived from the herb Radix Sophorae Flave,has been shown to ameliorate the clinical signs,inflammatory infiltration,demyelination in acute EAE rats.However,whether MAT protect from EAE by adjusting Th and Treg cells response in specific-cellular and molecular level is unknown.METHODS Herein,MAT was tested for its effects on Th1,Th2,Th17 and Treg cells in the spinal cord of EAE mice and splenocyte-extracted from EAE mice with MOG35-55-restimulated,respectively.RESULTS Our findings revealed that MAT significantly inhibit the proliferation of splenocyte,and remarkably down-regulate the differentiation of Th1/Th17 cells with decreased expressions of CD4+IFN-γ+cells and CD4+IL-17+cells in vivo and IL-17,IFN-γ,ROR-γt,T-bet in vitro,meanwhile it dramatically up-regulate the Th2/Treg cells response associated with increased levels of CD4+TGF-β+1cells and CD4+IL-10+cells in vivo and IL-4,IL-10,TGF-β1,Foxp3 and GATA3in vitro.CONCLUSION Considering the effective therapeutic effects of MAT on EAE,it′s worth to find its new values on other autoimmune diseases.
基金Supported by Methodist Research Institute,Indiana University Health
文摘AIM: To investigate the temporal onset and dynamic interplay of CD4+ T helper cell subsets in experimental autoimmune encephalomyelitis(EAE).METHODS: EAE was induced in C57BL/6 mice by immunization with myelin oligodendrocyte glycoprotein peptide p35-55. The clinical signs were scored and the tissue samples and immune cells isolated for analysis at different phases of EAE. The expression levels of inflammatory cytokines and related transcription factors were detected by quantitative reverse transcription polymerase chain reaction(PCR) and enzyme linked immunosorbant assay(ELISA). The percentages of Th1, Th17, Th2, Treg and memory T cell subsets in EAE were analyzed by immunostaining and flow cytometry.The data were analyzed by statistical techniques.RESULTS: Quantitative real-time PCR analysis showedthat EAE mice express elevated levels of Th1 [interferon gamma(IFNγ), interleukin(IL)-12p40 ], Th17 [IL-17, related orphan receptor gamma(RORγ), IL-12p40] and Treg [Foxp3, Epstein-Barr virus induced gene 3(EBI3), IL-10 ] genes in the central nervous system at the peak of the disease. Whereas, the expression of Th1(IFNγ, T-bet, IL-12p35, IL-12p40), Th17(RORγ, IL-12p40), Th2(IL-4) and Treg(Foxp3, EBI3) response genes was reduced in the spleen during pre-disease but gradually recovered at the later phases of EAE. ELISA and flow cytometry analyses showed an increase in Th17 response in the periphery, while Th1 response remained unchanged at the peak of disease. The m RNA levels of IFNγ, IL-17 and IL-12p40 in the brain were increased by 23(P < 0.001), 9(P < 0.05) and 14(P < 0.01) fold, respectively, on day 21 of EAE. Conversely, the mR NA expression of IL-10 was increased by 2 fold(P < 0.05) in the spleen on day 21. CD4+CD25+Foxp3+Treg response was reduced at pre-disease but recovered to naíve levels by disease onset. The percentage of CD25+Foxp3+ regulatory T cells decreased from 7.7% in the naíve to 3.2%(P < 0.05) on day 7 of EAE, which then increased to 8.4% by day 28. Moreover, the CD4+CD127+CD44high memory T cell response was increased during the onset and recovery phases of EAE. The memory and effector cells showed an inverse relationship in EAE, where the memory T cells increased from 12.3% in naive to 20% by day 21, and the effector cells decreased from 32% in naíve to 21%(P < 0.01) by day 21. The wild type C57BL/6 mice with EAE showed elevated levels of effector-memory T cells(TEM) with concomitant reduction in central-memory T cells(TCM), but the EAE-resistant IL-7R deficient mice showed elevated TCM with no effect on TEM cells in EAE.CONCLUSION: Our findings highlight the temporal onset and dynamic interplay of effector, memory and regulatory CD4+ T cell subsets and its significance to clinical outcome in EAE and other autoimmune diseases.
基金supported by grants from the National Institutes of Health(NS094151 and NS105689)the National Multiple Sclerosis Society(RG5239-A-3)(to WL)
文摘The transcription factor nuclear factor κB(NF-κB) plays major roles in inflammatory diseases through regulation of inflammation and cell viability.Multiple sclerosis(MS) is a chronic inflammatory demyelinating and neurodegenerative disease of the central nervous system(CNS).It has been shown that NF-κB is activated in multiple cell types in the CNS of MS patients,including T cells,microglia/macrophages,astrocytes,oligodendrocytes,and neurons.Interestingly,data from animal model studies,particularly studies of experimental autoimmune encephalomyelitis,have suggested that NF-κB activation in these individual cell types has distinct effects on the development of MS.In this review,we will cover the current literature on NF-κB and the evidence for its role in the development of MS and its animal model experimental autoimmune encephalomyelitis.
基金the National Natural Science Foundation of China,No.30672692
文摘<正>BACKGROUND:Yishendaluo decoction reduces production of inflammatory mediators,relieves damage due to inflammatory reactions,and improves neural functions during experimental autoimmune encephalomyelitis. OBJECTIVE:To investigate the effects of Yishendaluo decoction on a mouse model of experimental autoimmune encephalomyelitis. DESIGN,TIME AND SETTING:The randomized,controlled,neuropathological,and molecular biological animal study was performed at the Key Laboratory of Chinese Internal Medicine,Ministry of Education,Dongzhimen Hospital of Beijing University of Chinese Medicine and Center for Neuroinformatics,General Hospital of Chinese PLA from 2005 to 2006. MATERIALS:Yishendaluo decoction pieces consisting of prepared rehmannia root,colla cornus cervi,cape jasmine fruit,and grassleaf sweetflag rhizome were purchased from the Dongzhimen Hospital of Beijing University of Chinese Medicine.Rabbit anti-mouseβ-amyloid precursor protein and p38 polyclonal antibody(Zhongshan Goldenbridge Biotechnology,China),as well interferon-γand interleukin-4 ELISA kit(Boster,China),were used in this study. METHODS:A total of 96 healthy,female,SJL/J mice,aged 8-12 weeks,were equally and randomly assigned to normal,model,hormone,and Chinese medicine groups.A total of 0.2 mL antigen preparation,supplemented with 150μg PLP139-151 and 400μg H37RA,was subcutaneously injected into the upper abdomen of mice from the model,hormone,and Chinese medicine groups. Mouse models of experimental autoimmune encephalomyelitis were established by intravenous injection of 0.1 mL Bordetella pertussis solution containing 0.6×10~6 Bordetella pertussis at days 1 and 3.Mice from the model,Chinese medicine,and hormone groups were respectively subjected to 0.2 mL saline,2 g/kg Yishendaluo decoction,and 0.078 mg/kg prednisone acetate,once daily for 14 consecutive days.Mice from the normal group were left intact. MAIN OUTCOME MEASURES:Pathological changes were observed using hematoxylin-eosin staining and Luxol fast blue staining.Expression ofβ-amyloid precursor protein and p38 protein was determined by immunohistochemistry.Levels of interferon-γand interleukin-4 were detected by ELISA.Behavioral changes were assessed in mice according to scores of neurological function. RESULTS:A few inflammatory cell infiltration,nerve fiber breakage and slight demyelination were detected in the central nervous system of mice from the Chinese medicine and hormone groups compared with the model group.Expression ofβ-amyloid precursor protein and p38 protein was significantly diminished in the central nervous system of mice from the Chinese medicine and hormone groups compared with the model group(P<0.05 or P<0.01),and the decrease was greatest in the Chinese medicine group.The decrease in mouse weight was not significant,and neurological function scores were less in the Chinese medicine and hormone groups compared with the model group(P<0.05 or P<0.01).Interferon-γlevels were significantly reduced(P<0.01),and interleukin-4 levels were significantly increased(P<0.01) in the brains of the Chinese medicine and hormone groups,compared with the model group. CONCLUSION:Yishendaluo decoction improved neurological function in mice with experimental autoimmune encephalomyelitis by downregulatingβ-amyloid precursor protein expression,resisting axonal degeneration,and relieving inflammatory reaction.The anti-inflammatory mechanism was regulated by inhibition of the p38 mitogen-activated protein kinase signal pathway.
基金the National Natural Science Foundation of China,No. 30873230Beijing Natural Science Foundation,No. 7092014+1 种基金Scientific Research Common Program of Beijing Municipal Education Commission,No. KM2007100025015Fund-ing Project for Academic Human Resources Devel-opment in Institutions of Higher Learning Under the Jurisdiction of Beijing Mu-nicipality,No. PHR201008401
文摘Studies have demonstrated that amyloid precursor protein(APP)expression increases in multiple sclerosis tissues during acutely and chronically active stages.To determine the relationship between axonal injury and regeneration in multiple sclerosis,an animal model of experimental autoimmune encephalomyelitis was induced using different doses of myelin basic protein peptide.APP and growth-associated protein 43(GAP-43),which is considered a specific marker of neural regeneration,were assessed by western blot analysis.Expression of APP and GAP-43,as well as the correlation between these two proteins,in brain white matter and spinal cord tissues of experimental autoimmune encephalomyelitis rats at different pathological stages was analyzed.Results showed that APP and GAP-43 expression increased during the acute stage and decreased during remission,with a positive correlation between APP and GAP-43 expression in brain white matter and spinal cord tissues.These results suggest that APP and GAP-43 could provide nutritional and protective effects on damaged neurons.
文摘Multiple sclerosis(MS) is a common demyelinating central nervous system disease associated with progressive physical impairment. To study the mechanism underlying disease pathogenesis and develop potential treatments, experimental autoimmune encephalomyelitis(EAE) is often used as an animal model. EAE can be induced in various species by introducing specific antigens, which ultimately result in motor dysfunction. Although the severity of the paralysis is indicated using the EAE score, there is no standard scoring system for EAE signs, and there is variability between research groups with regard to the exact EAE scoring system utilized. Here, we describe the criteria used for EAE scoring systems in various laboratories and suggest combining EAE score with another quantitative index to evaluate paralysis, such as the traveled distance, with the goal of facilitating the study of the mechanisms and treatment of MS.
基金the National Natural Science Foundation of China, No. 30672692
文摘This study investigated the effects of Yishendaluo decoction on the loss of blood-brain barrier integrity in mice exhibiting experimental autoimmune encephalomyelitis.To this end,we used real-time fluorescent quantitative PCR to measure the levels of mRNAs specific to the T cell markers CD4 and CD8,and the monocyte marker CD11b.In addition,we used Evans blue dye extravasation in the spinal cord and brain tissues to assess blood-brain barrier permeability.The results indicated that an increase in blood-brain barrier permeability was associated with an increase in CD4,CD8 and CD11b mRNA expression in experimental autoimmune encephalomyelitis mice.Yishendaluo decoction administration significantly reversed inflammatory cell accumulation in cerebral tissues of experimental autoimmune encephalomyelitis mice.
基金supported by Multiple Sclerosis Research Australia and Trish Multiple Sclerosis Research Foundation Postgraduate Scholarship(to JYL)the National Multiple Sclerosis Society Project Grant#RG4398A1/1+2 种基金International Progressive Multiple Sclerosis Alliance Challenge Award#PA0065Multiple Sclerosis Research Australia and Trish Multiple Sclerosis Research Foundation#15-022Bethlehem Griffiths Research Foundation#BGRF1706(to SP)
文摘Myelin-associated inhibitory factors within the central nervous system(CNS) are considered to be one of the main obstacles for axonal regeneration following disease or injury. The nogo receptor 1(NgR1) has been well documented to play a key role in limiting axonal regrowth in the injured and diseased mammalian CNS. However, the role of nogo receptor in immune cell activation during CNS inflammation is yet to be mechanistically elucidated. Microglia/macrophages are immune cells that are regarded as pathogenic contributors to inflammatory demyelinating lesions in multiple sclerosis(MS). In this study, the animal model of MS, experimental autoimmune encephalomyelitis(EAE) was induced in ngr1^(+/+) and ngr1^(–/–) female mice following injection with the myelin oligodendrocyte glycoprotein(MOG_(35–55)) peptide. A fatemap analysis of microglia/macrophages was performed throughout spinal cord sections of EAE-induced mice at clinical scores of 0, 1, 2 and 3, respectively(increasing locomotor disability) from both genotypes, using the CD11 b and Iba1 cell markers. Western immunoblotting using lysates from isolated spinal cord microglia/macrophages, along with immunohistochemistry and flow cytometric analysis, was performed to demonstrate the expression of nogo receptor and its two homologs during EAE progression. Myelin protein engulfment during EAE progression in ngr1^(+/+) and ngr1^(–/–) mice was demonstrated by western immunblotting of lysates from isolated spinal cord microglia/macrophages, detecting levels of Nogo-A and MOG. The numbers of M1 and M2 microglia/macrophage phenotypes present in the spinal cords of EAE-induced ngr1^(+/+) and ngr1^(–/–) mice, were assessed by flow cytometric analysis using CD38 and Erg-2 markers. A significant difference in microglia/macrophage numbers between ngr1^(+/+) and ngr1^(–/–) mice was identified during the progression of the clinical symptoms of EAE, in the white versus gray matter regions of the spinal cord. This difference was unrelated to the expression of Ng R on these macrophage/microglial cells. We have identified that as EAE progresses, the phagocytic activity of microglia/macrophages with myelin debris, in ngr1^(–/–) mice, was enhanced. Moreover, we show a modulation from a predominant M1-pathogenic to the M2-neurotrophic cell phenotype in the ngr1^(–/–) mice during EAE progression. These findings suggest that CNS-specific macrophages and microglia of ngr1^(–/–) mice may exhibit an enhanced capacity to clear inhibitory molecules that are sequestered in inflammatory lesions.
基金the Key Combination Program of Capital Medical Development Foundation, No. 2005-SF-I-001the Natural Science Foundation of Beijing, No. 7102051+1 种基金Beijing Science and Technology Development Foundation of Traditional Chinese Medicine, No. JJ2009-27the National Natural Science Foundation of China, No. 81072765
文摘Zuogui pills have been shown to attenuate the inflammatory reaction in a rat model of experimental autoimmune encephalomyelitis(EAE).The present study attempted to investigate the pathology underlying the influence of Zuogui pills on myelinolysis in EAE rats.Hematoxylin-eosin and Luxol fast blue staining showed that the myelinolysis foci in the cerebrum,cerebellum,brain stem,and the spinal cord of EAE rats were significantly decreased,along with serum myelin basic protein content following treatment with Zuogui pills.
基金the Natural Science Foundation of Ministry of Science and Technology of China,No.30230140a grant from Merck Serono (China)
文摘The present study established a chronic experimental autoimmune encephalomyelitis model in C57BL/6 mice induced by myelin oligodendrocyte glycoprotein peptides and complete Freund's adjuvant.Onset latency was 12 days,with an incidence rate of 100%.Neuropathological characteristics included perivascular inflammatory cell infiltration,demyelination,neuronal degeneration,and axonal damage within cerebral and myelic white matter.Electron microscopy revealed swollen mitochondria,complete organ disappearance,and fused or broken myelin sheath structure,which were accompanied by myelin sheath reconstruction.Moreover,axonal damage was not consistent with demyelination distribution,and severity of axonal damage did not correlate with demyelination.Results suggested that axonal damage in an experimental autoimmune encephalomyelitis model is not secondary to inflammatory demyelination.
基金The project supported by National Natural Science Foundation of China(81341088,81001656)Beijing Nova Star Program in Science and Technology(Z12111000250000)
文摘OBJECTIVE In order to investigate whether cornel iridoid glycoside(CIG),the main component extracted from Cornus officinalis,can treat demyelinating diseases of the central nervous system(CNS)such as multiple sclerosis(MS).METHODS CIG(30,60 and 120mg·kg-1)or vehicle was intragastrically administered once daily to rats,starting immediately after purified myelin basic protein(MBP)68-86 peptides immunization until day 20 post immunization(p.i.).Histopathological staining,enzyme-linked immunosorbent assay,biochemical methods and Western blotting approaches were used to evaluate the disease incidence and severity,neuroinflammatory and neurotrophic response in the CNS.RESULTS Neurological deficit and proportion of incidence seen in EAE rats were significantly reduced by CIG treatment in a dose-dependent manner.Histopathological staining showed that CIG treatment alleviated demyelination and inflammatory infiltration,increased the number of oligodendrocytes,enhanced the expression of brain-derived neurotrophic factor(BDNF).Production of proinflammatory molecules such as interleukin-1β(IL-1β),tumour necrosis factor-αand interferon-γwere also inhibited by CIG administration.CIG could ameliorate phosphorylation of STAT1,STAT3 and JAK1 as well as IL-6/IL-6 Rexpression,which involved in immune response and inflammation.CONCLUSION Our results demonstrated that CIG may ameliorate EAE rats through down-regulation of JAK/STAT signaling pathway.This study gave new insight into the novel regulatory mechanism of CIG and highlight novel therapeutic targets and a potential therapeutic agent for the treatment of MS.
文摘Therapeutic administration of fingolimod hydrochloride (FTY720), the functional antagonist at sphingosine 1-phosphate (S1P) receptor 1 (S1P1) shows a marked improving effect on experimental autoimmune encephalomyelitis (EAE) induced by myelin oligodendrocyte glycoprotein (MOG) in C57BL/6 mice. However, this treatment showed an only partial inhibition of Th1/Th17 cell infiltration into the central nervous system (CNS), suggesting that down-regulation of lymphocytic S1P1 is insufficient to explain the therapeutic effect of FTY720 on EAE. On the other hand, the therapeutic administration of FTY720 reduced the mRNA expressions of IL-6, CCL2, and glial fibrillary acidic protein, an activation marker of astrocytes, in the CNS of EAE mice. In human astrocytic glyoma, U373MG cells, mRNA expression of S1P1 was higher as compared with those of the other S1P receptor subtypes and phosphorylation of Akt was induced by S1P, FTY720-phosphate (FTY720-P), or an S1P1-selective agonist, SEW2871. FTY720-P appeared to induce down-regulation of S1P1 in U373MG cells, implying a functional antagonism at S1P1 on astrocytes. S1P but not FTY720-P induced production of IL-6, IL-8, and CCL2 significantly and treatment with FTY720-P or SEW2871 inhibited production of these pro-inflammatory cytokines from U373MG cells stimulated with S1P. These results suggest that S1P-S1P1 axis induces production of pro-inflammatory cytokines by astrocytes. Consequently, it is highly probable that the therapeutic effects of FTY720 on EAE are caused by inhibiting not only egress of myelin-specific Th cells from the draining lymph nodes but also activation of astrocytes in the CNS.
文摘Infiltration of myelin-specific helper T (Th) cells into the central nervous system (CNS) plays a key role in pathogenesis of experimental autoimmune encephalomyelitis (EAE). In this study, we investigated the involvement of sphingosine 1-phosphate (S1P)-S1P receptor 1 (S1P1) axis in lymphocytes for EAE development when C57BL/6 mice were immunized with myelin oliogodendrocyte glycoprotein (MOG). The expression of S1P1 mRNA and S1P responsiveness of lymphocytes in draining lymph nodes (DLN) were down-regulated markedly after MOG immunization until onset of EAE. Accompanying with reacquisition of down-regulated S1P1 transcript and S1P responsiveness in DLN lymphocytes, MOG-immunized mice developed EAE symptoms with significant infiltration of Th1 and Th17 cells into the CNS and a marked elevation of IFN-γ, T-bet, IL-17, and RORγt mRNA expressions. Prophylactic administration of an S1P1 functional antagonist, fingolimod hydrochloride (FTY720, 0.3 mg/kg, orally) significantly inhibited EAE development and almost completely prevented infiltration of Th1 and Th17 cells into the CNS with a marked reduction of IFN-γ, T-bet, IL-17, and RORγt mRNA expressions. Similar results were obtained by treatment with an S1P1-selective agonist, SEW2871 or an S1P lyase inhibitor, 2-acetyl-4-tetrahydroxybutylimidazole. Moreover, FTY720-phosphate and SEW2871 inhibited in vitro migration of Th1 and Th17 cells toward S1P but did not affect cytokine production or generation of Th1 or Th17 cells. These results suggest that reacquisition of S1P1 expression in DLN lymphocytes plays a major role in trafficking of myelin antigen-specific Th1/Th17 cells from DLN to the CNS in EAE and that prophylactic effect of FTY720 on EAE is predominantly caused by functional antagonism via lymphocytic S1P1.
文摘AIM: To study the effect of blocking the eo-2 pathwayon the development and severity of experimental autoimmune encephalomyelitis(EAE). METHODS: We produced m Ab directed against eo-2,named D8. MOG35-55 induced-EAE mice were dailyintravenously injected with either 25 μg or 100 μg D8,or with vehicle control alone [phosphate-buffered saline(PBS)], starting from day 0 post immunization and weremonitored for EAE clinical score(n = 10 in each group).Mice were sacrificed on day 58 and their sera were assessed for the presence of anti-myelin oligodendrocyteglycoprotein(anti-MOG) antibodies autoantibodies, aswell as for the profile of pro-inflammatory cytokines andchemokines. Histological analysis of brain sections wasperformed by hematoxylin and eosin staining.RESULTS: Daily treatment of EAE induced mice with D8 significantly decreased the severity of EAE symptoms. Treatment with both concentrations of D8 ameliorated EAE symptoms compared to PBS treated mice, starting from day 42 post immunization(0.89 ± 0.35 in D8 25 μg and D8 100 μg treated groups vs 2.11 ± 0.38 in the PBS treated group, P = 0.03). A significant improvement in EAE clinical score compared to total Ig G treated mice was observed with the higher concentration of D8(0.81 ± 0.38 in D8 100 μg treated group vs 2.11 ± 0.31 in Ig G1 treated group, on day 56 post immunization, P = 0.04). D8 treated mice with EAE did not significantly exhibit lower sera levels of anti-MOG autoantibodies compared to Ig G-treated mice. However, they expressed lower sera levels of the pro-inflammatory cytokines: tumor necrosis factor(7.8 ± 0.2 pg/m L in D8 100 μg treated mice vs 19.9 ± 3.4 pg/m L in Ig G treated mice, P = 0.005) and interferon-gamma(1.4 ± 0.6 pg/m L in D8 100 μg treated mice vs 3.6 ± 0.4 pg/m L in Ig G treated mice, P = 0.02), as well as reduced levels of the chemokine macrophage chemoattractant protein-1(27.2 ± 3.1 pg/m L in D8 100 μg treated mice vs 63.7 ± 12.3 pg/m L in Ig G treated mice, P = 0.03). These findings indicate that blocking the eo-2 pathway in EAE may affect not only eosinophil infiltration into the central nervous system(CNS), but also have an effect on monocytes and T cells, but not humoral, mediated responses. Histological analysis of the brains of D8 treated mice with EAE support that this treatment decreases immune cells infiltrates in the CNS.CONCLUSION: Taken together, these findings suggest a role for eo-2 in EAE pathogenesis and consequentially may support a therapeutic potential of anti-eo-2 neutralizing m Ab in multiple sclerosis.
