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
Monocytes are key effectors in autoimmunity-related diseases in the central nervous system(CNS)due to the critical roles of these cells in the production of proinflammatory cytokines,differentiation of T-helper(Th)cel...Monocytes are key effectors in autoimmunity-related diseases in the central nervous system(CNS)due to the critical roles of these cells in the production of proinflammatory cytokines,differentiation of T-helper(Th)cells,and antigen presentation.The JAK-STAT signaling is crucial for initiating monocytes induced immune responses by relaying cytokines signaling.However,the role of this pathway in modulating the communication between monocytes and Th cells in the pathogenesis of multiple sclerosis(MS)is unclear.Here,we show that the JAK1/2/3 and STAT1/3/5/6 subtypes involved in the demyelination mediated by the differentiation of pathological Th1 and Th17 and the CNS-infiltrating inflammatory monocytes in experimental autoimmune encephalomyelitis(EAE),a model for MS.JAK inhibition prevented the CNS-infiltrating CCR2-dependent Ly6^(Chi)monocytes and monocyte-derived dendritic cells in EAE mice.In parallel,the proportion of GM-CSF^(+)CD4^(+)T cells and GM-CSF secretion were decreased in pathological Th17 cells by JAK inhibition,which in turns converted CNS-invading monocytes into antigen-presenting cells to mediate tissue damage.Together,our data highlight the therapeutic potential of JAK inhibition in treating EAE by blocking the GM-CSF-driven inflammatory signature of monocytes.展开更多
Objective: To examine the anti-inflammatory effect of grape seed extract(GSE) in animal and cellular models and explore its mechanism of action. Methods: This study determined the inhibitory effect of GSE on macrophag...Objective: To examine the anti-inflammatory effect of grape seed extract(GSE) in animal and cellular models and explore its mechanism of action. Methods: This study determined the inhibitory effect of GSE on macrophage inflammation and Th1 and Th17 polarization in vitro. Based on the in vitro results, the effects and mechanisms of GSE on multiple sclerosis(MS)-experimental autoimmune encephalomyelitis(EAE) mice model were further explored. The C57BL/6 mice were intragastrically administered with 50 mg/kg of GSE once a day from the 3rd day to the 27th day after immunization. The activation of microglia, the polarization of Th1 and Th17and the inflammatory factors such as tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), IL-6, IL-12, IL-17and interferon-γ(IFN-γ) secreted by them were detected in vitro and in vivo by flow cytometry, enzyme linked immunosorbent assay(ELISA), immunofluorescence staining and Western blot, respectively. Results: GSE reduced the secretion of TNF-α, IL-1β and IL-6 in bone marrow-derived macrophages stimulated by lipopolysaccharide(P<0.01), inhibited the secretion of TNF-α, IL-1β, IL-6, IL-12, IL-17 and IFN-γ in spleen cells of EAE mice immunized for 9 days(P<0.05 or P<0.01), and reduced the differentiation of Th1 and Th17 mediated by CD3 and CD28 factors(P<0.01). GSE significantly improved the clinical symptoms of EAE mice, and inhibited spinal cord demyelination and inflammatory cell infiltration. Peripherally, GSE downregulated the expression of toll-like-receptor 4(TLR4) and Rho-associated kinase(ROCKⅡ, P<0.05 or P<0.01), and inhibited the secretion of inflammatory factors(P<0.01 or P<0.05). In the central nervous system, GSE inhibited the infiltration of CD45+CD11b+and CD45+CD4+cells, and weakened the differentiation of Th1 and Th17(P<0.05). Moreover, it reduced the secretion of inflammatory factors(P<0.01), and prevented the activation of microglia(P<0.05). Conclusion: GSE had a beneficial effect on the pathogenesis and progression of EAE by inhibiting inflammatory response as a potential drug and strategy for the treatment of MS.展开更多
Multiple sclerosis(MS)is a chronic and incurable autoimmune neurodegenerative disease of the central nervous system.Although the symptoms of MS can be managed by vitamin D3 treatment alone,this condition cannot be com...Multiple sclerosis(MS)is a chronic and incurable autoimmune neurodegenerative disease of the central nervous system.Although the symptoms of MS can be managed by vitamin D3 treatment alone,this condition cannot be completely eradicated.Thus,there might be unknown factors capable of regulating the vitamin D receptor(VDR).Genome-wide analysis showed that miRNAs were associated with VDRs.We sought to determine the role and mechanism of action of miRNA-125a-5p and VDRs in a model of MS,mice with experimental autoimmune encephalomyelitis(EAE),which was induced by myelin oligodendrocyte glycoprotein 35–55 peptides.EAE mice showed decreased mean body weight but increased mean clinical scores compared with vehicle or control mice.And inflammatory infiltration was found in the lumbosacral spinal cord of EAE mice.In addition,VDR expression was significantly lower while the expression of miR-125a-5p was markedly higher in the spinal ventral horn of EAE mice than in vehicle or control mice.Importantly,activation of VDRs by paricalcitol or inhibition of miR-125a-5p by its antagomir markedly decreased the mean clinical scores in EAE mice.Interestingly,VDR and miR-125a-5p were co-localized in the same neurons of the ventral horn.More importantly,inhibition of miR-125a-5p remarkably blocked the decrease of VDRs in EAE mice.These results support a critical role for miR-125a-5p in modulating VDR activity in EAE and suggest potential novel therapeutic interventions.展开更多
The present study was conducted to clarify the therapeutic effect of cornuside on experimental autoimmune encephalomyelitis(EAE)and its influence on T helper 17(Th17)cell and regulatory T(Treg)cell infiltration into t...The present study was conducted to clarify the therapeutic effect of cornuside on experimental autoimmune encephalomyelitis(EAE)and its influence on T helper 17(Th17)cell and regulatory T(Treg)cell infiltration into the central nervous system.Rats were randomly placed into four treatment groups:control,EAE,EAE+cornuside,and EAE+prednisolone.The neurological function scores of rats were assessed daily.On the second day after EAE rats began to show neurological deficit symptoms,the four groups were treated with normal saline,normal saline,cornuside(150 mg/kg),and prednisolone(5 mg/kg),respectively.The treatment was discontinued after two weeks,and the spinal cord was obtained for hematoxylin and eosin(H&E)and luxol fast blue staining,as well as retinoic acid receptor-related orphan receptorγ(RORγ)and forkhead box protein P3(Foxp3)immunohistochemical staining.Blood was collected for Th17 and Treg cell flow cytometry testing,and the serum levels of interleukin(IL)-17A,IL-10,transforming growth factor-β(TGF-β),IL-6,IL-23,and IL-2 were measured via enzymelinked immunosorbent assay(ELISA).Compared with rats in the EAE group,rats in the EAE+cornuside and EAE+prednisolone groups began to recover from neurological deficits earlier,and had a greater degree of improvement of symptoms.Focal inflammation,demyelination,and RORγ-positive cell infiltration were reduced by cornuside or prednisolone treatment,whereas the Foxp3-positive cell numbers were not significantly different.Meanwhile,the number of Th17 cells and the IL-17A,IL-6,and IL-23 levels were lower in the blood after cornuside or prednisolone treatment,whereas the number of Treg cells or the levels of IL-10,TGF-β,and IL-2 were not markedly different.Cornuside can alleviate symptoms of EAE neurological deficits through its anti-inflammatory and immunosuppressive effects,and Th17 cells may be one of its therapeutic targets.展开更多
Circular RNAs(circRNAs)regulate gene expression and participate in various biological and pathological processes.However,little is known about the effects of specific circRNAs on T helper cell 17(Th17)differentiation ...Circular RNAs(circRNAs)regulate gene expression and participate in various biological and pathological processes.However,little is known about the effects of specific circRNAs on T helper cell 17(Th17)differentiation and related autoimmune diseases,such as multiple sclerosis(MS).Here,using transcriptome microarray analysis at different stages of experimental autoimmune encephalomyelitis(EAE),we identified the EAE progression-related circINPP4B,which showed upregulated expression in Th17 cells from mice with EAE and during Th17 differentiation in vitro.Silencing of circINPP4B inhibited Th17 differentiation and alleviated EAE,characterized by less demyelination and Th17 infiltration in the spinal cord.Mechanistically,circINPP4B served as a sponge that directly targeted miR-30a to regulate Th17 differentiation.Furthermore,circINPP4B levels were associated with the developing phases of clinical relapsing-remitting MS patients.Our results indicate that circINPP4B plays an important role in promoting Th17 differentiation and progression of EAE by targeting miR-30a,which provides a potential diagnostic and therapeutic target for Th17-mediated MS.展开更多
Background:Huangqi Guizhi Wuwu decoction(HQGZWW)exhibits good effects when administered to treat multiple sclerosis(MS)and its animal model,experimental autoimmune encephalomyelitis(EAE).Understanding the precise mech...Background:Huangqi Guizhi Wuwu decoction(HQGZWW)exhibits good effects when administered to treat multiple sclerosis(MS)and its animal model,experimental autoimmune encephalomyelitis(EAE).Understanding the precise mechanism of this decoction is thus important.Based on the findings of our previous study,the aim of the present study was to understand the role of antigen-specific CD8^(+)T-cells on the pathogene sis of MS/EAE when HQGZWW is administered as treatment.Methods:Myelin oligodendrocyte glycoprotein(MOG);-induced mice were administered distilled water,prednisone,and high dose or low dose HQGZWW.After purified CD4^(+)and CD8^(+)T-cells were stimulated with the MOG;peptide,proliferation and cytokine secretion assays were performed.To establish the adoptive transfer EAE model,naive mice were injected with MOG;-CD8^(+)or CD4^(+)T-cells.Results:Significant improvements in EAE score and pathology were observed in the high dose HQGZWW and prednisone groups.Compared to the low dose HQGZWW and distilled water groups,lower antigen-specific re sponses,lower levels of interferon-gamma,and higher levels of interleukin(IL)-4 and IL-10 from CD8^(+)and CD4^(+)T cells were observed in the high dose HQGZWW and prednisone groups.Finally,the EAE score was observed to be similar between the high dose HQGZWW group and prednisone group;however,this finding was not observed in the low dose HQGZWW group.Conclusion:Our findings suggest that high dose HQGZWW has similar effects on cell proliferation,cytokine secretion,and EAE score to prednisone,while low dose HQGZWW does not have such effect.The protective role of HQGZWW against EAE might thus depend on the Th2 cytokine secretion profile induced by either MOG;specific CD8^(+)or CD4^(+)T-cells.展开更多
The lack of targeted and high-efficiency drug delivery to the central nervous system(CNS)nidus is the main problem in the treatment of demyelinating disease.Extracellular vesicles(EVs)possess great promise as a drug d...The lack of targeted and high-efficiency drug delivery to the central nervous system(CNS)nidus is the main problem in the treatment of demyelinating disease.Extracellular vesicles(EVs)possess great promise as a drug delivery vector given their advanced features.However,clinical applications are limited because of their inadequate targeting ability and the“dilution effects”after systemic administration.Neural stem cells(NSCs)supply a plentiful source of EVs on account of their extraordinary capacity for self-renewal.Here,we have developed a novel therapeutic system using EVs from modified NSCs with high expressed ligand PDGF-A(EVPs)and achieve local delivery.It has been demonstrated that EVPs greatly enhance the target capability on oligodendrocyte lineage.Moreover,EVPs are used for embedding triiodothyronine(T3),a thyroid hormone that is critical for oligodendrocyte development but has serious side effects when systemically administered.Our results demonstrated that systemic injection of EVPs+T3,versus EVPs or T3 administration individually,markedly alleviated disease development,enhanced oligodendrocyte survival,inhibited myelin damage,and promoted myelin regeneration in the lesions of experimental autoimmune encephalomyelitis mice.Taken together,our findings showed that engineered EVPs possess a remarkable CNS lesion targeting potential that offers a potent therapeutic strategy for CNS demyelinating diseases as well as neuroinflammation.展开更多
The PIK3C3/VPS34 subunit of the class III phosphatidylinositol 3-kinase(PtdIns3K)complex plays a role in both canonical and noncanonical autophagy,key processes that control immune-cell responsiveness to a variety of ...The PIK3C3/VPS34 subunit of the class III phosphatidylinositol 3-kinase(PtdIns3K)complex plays a role in both canonical and noncanonical autophagy,key processes that control immune-cell responsiveness to a variety of stimuli.Our previous studies found that PIK3C3 is a critical regulator that controls the development,homeostasis,and function of dendritic and T cells.In this study,we investigated the role of PIK3C3 in myeloid cell biology using myeloid cell-specific Pik3c3-deficient mice.We found that Pik3c3-deficient macrophages express increased surface levels of major histocompatibility complex(MHC)class I and class II molecules.In addition,myeloid cell-specific Pik3c3 ablation in mice caused a partial impairment in the homeostatic maintenance of macrophages expressing the apoptotic cell uptake receptor TIM-4.Pik3c3 deficiency caused phenotypic changes in myeloid cells that were dependent on the early machinery(initiation/nucleation)of the classical autophagy pathway.Consequently,myeloid cell-specific Pik3c3-deficient animals showed significantly reduced severity of experimental autoimmune encephalomyelitis(EAE),a primarily CD4^(^(+))T-cell-mediated mouse model of multiple sclerosis(MS).This disease protection was associated with reduced accumulation of myelin-specific CD4^(^(+))T cells in the central nervous system and decreased myeloid cell IL-1βproduction.Further,administration of SAR405,a selective PIK3C3 inhibitor,delayed disease progression.Collectively,our studies establish PIK3C3 as an important regulator of macrophage functions and myeloid cell-mediated regulation of EAE.Our findings also have important implications for the development of small-molecule inhibitors of PIK3C3 as therapeutic modulators of MS and other autoimmune diseases.展开更多
Objective To investigate the effets of nordihydroguaiaretic acid(NDGA)on the expression of IL-6,IL-17and TGF-βin mice with experimental autoimmune encephalomyelitis(EAE).Methods C57BL/6 mice were immunized with MOG35...Objective To investigate the effets of nordihydroguaiaretic acid(NDGA)on the expression of IL-6,IL-17and TGF-βin mice with experimental autoimmune encephalomyelitis(EAE).Methods C57BL/6 mice were immunized with MOG35-55 to induce EAE.The 54 mice were randomly and equally divided into control group,model group and treatment group.Mice in treatment展开更多
Early growth response protein 1(Egr-1)triggers the transcription of many genes involved in cell growth,differentiation,synaptic plasticity,and neurogenesis.However,its mechanism in neuronal survival and degeneration i...Early growth response protein 1(Egr-1)triggers the transcription of many genes involved in cell growth,differentiation,synaptic plasticity,and neurogenesis.However,its mechanism in neuronal survival and degeneration is still poorly understood.This study demonstrated that Egr-1 was down-regulated at mRNA and protein levels in the central nervous system(CNS)of experimental autoimmune encephalomyelitis(EAE)mice.Egr-1 knockout exacerbated EAE progression in mice,as shown by increased disease severity and incidence;it also aggravated neuronal apoptosis,which was associated with weakened activation of the BDNF/TGFβ1/MAPK/Akt signaling pathways in the CNS of EAE mice.Consistently,Egr-1 siRNA promoted apoptosis but mitigated the activation of BDNF/TGFβ1/MAPK/Akt signaling in SH-SY5Y cells.Our results revealed that Egr-1 is a crucial regulator of neuronal survival in EAE by regulating TGFβ1-mediated signaling activation,implicating the important role of Egr-1 in the pathogenesis of multiple sclerosis as a potential novel therapy target.展开更多
Multiple sclerosis is a chronic autoimmune disease of the central nervous system and is generally considered to be a non-traumatic,physically debilitating neurological disorder.In addition to experiencing motor disabi...Multiple sclerosis is a chronic autoimmune disease of the central nervous system and is generally considered to be a non-traumatic,physically debilitating neurological disorder.In addition to experiencing motor disability,patients with multiple sclerosis also experience a variety of nonmotor symptoms,including cognitive deficits,anxiety,depression,sensory impairments,and pain.However,the pathogenesis and treatment of such non-motor symptoms in multiple scle rosis are still under research.Preclinical studies for multiple sclerosis benefit from the use of disease-appropriate animal models,including experimental autoimmune encephalomyelitis.Prior to understanding the pathophysiology and developing treatments for non-motor symptoms,it is critical to chara cterize the animal model in terms of its ability to replicate certain non-motor features of multiple sclerosis.As such,no single animal model can mimic the entire spectrum of symptoms.This review focuses on the non-motor symptoms that have been investigated in animal models of multiple sclerosis as well as possible underlying mechanisms.Further,we highlighted gaps in the literature to explain the nonmotor aspects of multiple sclerosis in expe rimental animal models,which will serve as the basis for future studies.展开更多
Structural plasticity is critical for the functional diversity of neurons in the brain.Experimental autoimmune encephalomyelitis(EAE)is the most commonly used model for multiple sclerosis(MS),successfully mimicking it...Structural plasticity is critical for the functional diversity of neurons in the brain.Experimental autoimmune encephalomyelitis(EAE)is the most commonly used model for multiple sclerosis(MS),successfully mimicking its key pathological features(inflammation,demyelination,axonal loss,and gliosis)and clinical symptoms(motor and non-motordysfunctions).Recentstudieshave demonstrated the importance of synaptic plasticity in EAE pathogenesis.In the present study,we investigated the features of behavioral alteration and hippocampal structural plasticity in EAE-affected mice in the early phase(11 days post-immunization,DPI)and chronic phase(28DPI).EAE-affected mice exhibited hippocampus-related behavioral dysfunction in the open field test during both early and chronic phases.Dendritic complexity was largely affected in the cornu ammonis 1(CA1)and CA3 apical and dentate gyrus(DG)subregions of the hippocampus during the chronic phase,while this effect was only noted in the CA1 apical subregion in the early phase.Moreover,dendritic spine density was reduced in the hippocampal CA1 and CA3 apical/basal and DG subregions in the early phase of EAE,but only reduced in the DG subregion during the chronic phase.Furthermore,mRNA levels of proinflammatory cytokines(Il1β,Tnfα,and Ifnγ)and glial cell markers(Gfap and Cd68)were significantly increased,whereas the expression of activity-regulated cytoskeletonassociated protein(ARC)was reduced during the chronic phase.Similarly,exposure to the aforementioned cytokines in primary cultures of hippocampal neurons reduced dendritic complexity and ARC expression.Primary cultures of hippocampal neurons also showed significantly reduced extracellular signal-regulated kinase(ERK)phosphorylation upon treatment with proinflammatory cytokines.Collectively,these results suggest that autoimmune neuroinflammation alters structural plasticity in the hippocampus,possibly through the ERK-ARC pathway,indicating that this alteration may be associated with hippocampal dysfunctions in EAE.展开更多
Hydroxycitric acid(HCA) is derived primarily from the Garcinia plant and is widely used for its anti-inflammatory effects. Multiple sclerosis can cause an inflammatory demyelination and axonal damage. In this study, t...Hydroxycitric acid(HCA) is derived primarily from the Garcinia plant and is widely used for its anti-inflammatory effects. Multiple sclerosis can cause an inflammatory demyelination and axonal damage. In this study, to validate the hypothesis that HCA exhibits therapeutic effects on multiple sclerosis, we established female C57BL/6 mouse models of multiple sclerosis, i.e., experimental autoimmune encephalomyelitis,using Complete Freund's Adjuvant(CFA) emulsion containing myelin oligodendrocyte glycoprotein(35–55). Treatment with HCA at 2 g/kg/d for 3 weeks obviously improved the symptoms of nerve injury of experimental autoimmune encephalomyelitis mice, decreased serum interleulin-6, tumor necrosis factor alpha, nitric oxide, and malondialdehyde levels, and increased superoxide dismutase and glutathione reductase activities. These findings suggest that HCA exhibits neuroprotective effects on multiple sclerosis-caused nerve injury through ameliorating inflammation and oxidative stress.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 the National Natural Science Foundation of China(82293684,82293680,82104189,82273929)the National Key R&D Program of China(2020YFA0908004)+1 种基金CAMS Innovation Fund for Medical Science(2021-I2M-1-028,2022-I2M-2-002,2022-I2M-1-014,China)Special Research Fund for Central Universities,Peking Union Medical College grant(3332022146,China)。
文摘Monocytes are key effectors in autoimmunity-related diseases in the central nervous system(CNS)due to the critical roles of these cells in the production of proinflammatory cytokines,differentiation of T-helper(Th)cells,and antigen presentation.The JAK-STAT signaling is crucial for initiating monocytes induced immune responses by relaying cytokines signaling.However,the role of this pathway in modulating the communication between monocytes and Th cells in the pathogenesis of multiple sclerosis(MS)is unclear.Here,we show that the JAK1/2/3 and STAT1/3/5/6 subtypes involved in the demyelination mediated by the differentiation of pathological Th1 and Th17 and the CNS-infiltrating inflammatory monocytes in experimental autoimmune encephalomyelitis(EAE),a model for MS.JAK inhibition prevented the CNS-infiltrating CCR2-dependent Ly6^(Chi)monocytes and monocyte-derived dendritic cells in EAE mice.In parallel,the proportion of GM-CSF^(+)CD4^(+)T cells and GM-CSF secretion were decreased in pathological Th17 cells by JAK inhibition,which in turns converted CNS-invading monocytes into antigen-presenting cells to mediate tissue damage.Together,our data highlight the therapeutic potential of JAK inhibition in treating EAE by blocking the GM-CSF-driven inflammatory signature of monocytes.
基金Supported by the National Natural Science Foundation of China (No.81903596)Science and Technology Innovation Project of Shanxi Colleges (No.2019L0728)+2 种基金Leading Team of Medical Science and Technology of Shanxi Province (No.2020TD05)Funds for Construction of Key Disciplines from Shanxi University of Chinese Medicine (No.030200117)Cultivation Project of Shanxi University of Chinese Medicine (Nos.2019PY130 and 2020PY-JC-14)。
文摘Objective: To examine the anti-inflammatory effect of grape seed extract(GSE) in animal and cellular models and explore its mechanism of action. Methods: This study determined the inhibitory effect of GSE on macrophage inflammation and Th1 and Th17 polarization in vitro. Based on the in vitro results, the effects and mechanisms of GSE on multiple sclerosis(MS)-experimental autoimmune encephalomyelitis(EAE) mice model were further explored. The C57BL/6 mice were intragastrically administered with 50 mg/kg of GSE once a day from the 3rd day to the 27th day after immunization. The activation of microglia, the polarization of Th1 and Th17and the inflammatory factors such as tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), IL-6, IL-12, IL-17and interferon-γ(IFN-γ) secreted by them were detected in vitro and in vivo by flow cytometry, enzyme linked immunosorbent assay(ELISA), immunofluorescence staining and Western blot, respectively. Results: GSE reduced the secretion of TNF-α, IL-1β and IL-6 in bone marrow-derived macrophages stimulated by lipopolysaccharide(P<0.01), inhibited the secretion of TNF-α, IL-1β, IL-6, IL-12, IL-17 and IFN-γ in spleen cells of EAE mice immunized for 9 days(P<0.05 or P<0.01), and reduced the differentiation of Th1 and Th17 mediated by CD3 and CD28 factors(P<0.01). GSE significantly improved the clinical symptoms of EAE mice, and inhibited spinal cord demyelination and inflammatory cell infiltration. Peripherally, GSE downregulated the expression of toll-like-receptor 4(TLR4) and Rho-associated kinase(ROCKⅡ, P<0.05 or P<0.01), and inhibited the secretion of inflammatory factors(P<0.01 or P<0.05). In the central nervous system, GSE inhibited the infiltration of CD45+CD11b+and CD45+CD4+cells, and weakened the differentiation of Th1 and Th17(P<0.05). Moreover, it reduced the secretion of inflammatory factors(P<0.01), and prevented the activation of microglia(P<0.05). Conclusion: GSE had a beneficial effect on the pathogenesis and progression of EAE by inhibiting inflammatory response as a potential drug and strategy for the treatment of MS.
基金supported by the International Science and Technology Cooperation Plan of Guizhou Province,China[(2013)7027]the National Natural Science Foundation of China(81471137 and 31730040).
文摘Multiple sclerosis(MS)is a chronic and incurable autoimmune neurodegenerative disease of the central nervous system.Although the symptoms of MS can be managed by vitamin D3 treatment alone,this condition cannot be completely eradicated.Thus,there might be unknown factors capable of regulating the vitamin D receptor(VDR).Genome-wide analysis showed that miRNAs were associated with VDRs.We sought to determine the role and mechanism of action of miRNA-125a-5p and VDRs in a model of MS,mice with experimental autoimmune encephalomyelitis(EAE),which was induced by myelin oligodendrocyte glycoprotein 35–55 peptides.EAE mice showed decreased mean body weight but increased mean clinical scores compared with vehicle or control mice.And inflammatory infiltration was found in the lumbosacral spinal cord of EAE mice.In addition,VDR expression was significantly lower while the expression of miR-125a-5p was markedly higher in the spinal ventral horn of EAE mice than in vehicle or control mice.Importantly,activation of VDRs by paricalcitol or inhibition of miR-125a-5p by its antagomir markedly decreased the mean clinical scores in EAE mice.Interestingly,VDR and miR-125a-5p were co-localized in the same neurons of the ventral horn.More importantly,inhibition of miR-125a-5p remarkably blocked the decrease of VDRs in EAE mice.These results support a critical role for miR-125a-5p in modulating VDR activity in EAE and suggest potential novel therapeutic interventions.
基金This work was supported by the Traditional Chinese Medical Science and Technology Project of Zhejiang Province(No.2019ZA063)the Scientific Research Fund of Zhejiang Chinese Medical University(No.2019ZY09),China.
文摘The present study was conducted to clarify the therapeutic effect of cornuside on experimental autoimmune encephalomyelitis(EAE)and its influence on T helper 17(Th17)cell and regulatory T(Treg)cell infiltration into the central nervous system.Rats were randomly placed into four treatment groups:control,EAE,EAE+cornuside,and EAE+prednisolone.The neurological function scores of rats were assessed daily.On the second day after EAE rats began to show neurological deficit symptoms,the four groups were treated with normal saline,normal saline,cornuside(150 mg/kg),and prednisolone(5 mg/kg),respectively.The treatment was discontinued after two weeks,and the spinal cord was obtained for hematoxylin and eosin(H&E)and luxol fast blue staining,as well as retinoic acid receptor-related orphan receptorγ(RORγ)and forkhead box protein P3(Foxp3)immunohistochemical staining.Blood was collected for Th17 and Treg cell flow cytometry testing,and the serum levels of interleukin(IL)-17A,IL-10,transforming growth factor-β(TGF-β),IL-6,IL-23,and IL-2 were measured via enzymelinked immunosorbent assay(ELISA).Compared with rats in the EAE group,rats in the EAE+cornuside and EAE+prednisolone groups began to recover from neurological deficits earlier,and had a greater degree of improvement of symptoms.Focal inflammation,demyelination,and RORγ-positive cell infiltration were reduced by cornuside or prednisolone treatment,whereas the Foxp3-positive cell numbers were not significantly different.Meanwhile,the number of Th17 cells and the IL-17A,IL-6,and IL-23 levels were lower in the blood after cornuside or prednisolone treatment,whereas the number of Treg cells or the levels of IL-10,TGF-β,and IL-2 were not markedly different.Cornuside can alleviate symptoms of EAE neurological deficits through its anti-inflammatory and immunosuppressive effects,and Th17 cells may be one of its therapeutic targets.
基金This research was supported by the National Natural Science Foundation of China(81771337,81271345)The National Key R&D Program of China(2017YFA0104202)+3 种基金The Natural Science Foundation of Jiangsu Province(BK20161174)the 333 Project of Jiangsu ProvinceThe Xuzhou Basic Research Science and Technology Project(KC19059)Xuzhou Medical University Scientific Research Fund for Talents.
文摘Circular RNAs(circRNAs)regulate gene expression and participate in various biological and pathological processes.However,little is known about the effects of specific circRNAs on T helper cell 17(Th17)differentiation and related autoimmune diseases,such as multiple sclerosis(MS).Here,using transcriptome microarray analysis at different stages of experimental autoimmune encephalomyelitis(EAE),we identified the EAE progression-related circINPP4B,which showed upregulated expression in Th17 cells from mice with EAE and during Th17 differentiation in vitro.Silencing of circINPP4B inhibited Th17 differentiation and alleviated EAE,characterized by less demyelination and Th17 infiltration in the spinal cord.Mechanistically,circINPP4B served as a sponge that directly targeted miR-30a to regulate Th17 differentiation.Furthermore,circINPP4B levels were associated with the developing phases of clinical relapsing-remitting MS patients.Our results indicate that circINPP4B plays an important role in promoting Th17 differentiation and progression of EAE by targeting miR-30a,which provides a potential diagnostic and therapeutic target for Th17-mediated MS.
基金supported by Key Plans of Hunan Administration Traditional Chinese Medicine(No.201915 to YP)the grants from the Natural Science Foundation of Hunan Province.China(No.2018JJ6043 to YP)the Health and Family Plans commission of Hunan Province,China(No.B20180815to YP)。
文摘Background:Huangqi Guizhi Wuwu decoction(HQGZWW)exhibits good effects when administered to treat multiple sclerosis(MS)and its animal model,experimental autoimmune encephalomyelitis(EAE).Understanding the precise mechanism of this decoction is thus important.Based on the findings of our previous study,the aim of the present study was to understand the role of antigen-specific CD8^(+)T-cells on the pathogene sis of MS/EAE when HQGZWW is administered as treatment.Methods:Myelin oligodendrocyte glycoprotein(MOG);-induced mice were administered distilled water,prednisone,and high dose or low dose HQGZWW.After purified CD4^(+)and CD8^(+)T-cells were stimulated with the MOG;peptide,proliferation and cytokine secretion assays were performed.To establish the adoptive transfer EAE model,naive mice were injected with MOG;-CD8^(+)or CD4^(+)T-cells.Results:Significant improvements in EAE score and pathology were observed in the high dose HQGZWW and prednisone groups.Compared to the low dose HQGZWW and distilled water groups,lower antigen-specific re sponses,lower levels of interferon-gamma,and higher levels of interleukin(IL)-4 and IL-10 from CD8^(+)and CD4^(+)T cells were observed in the high dose HQGZWW and prednisone groups.Finally,the EAE score was observed to be similar between the high dose HQGZWW group and prednisone group;however,this finding was not observed in the low dose HQGZWW group.Conclusion:Our findings suggest that high dose HQGZWW has similar effects on cell proliferation,cytokine secretion,and EAE score to prednisone,while low dose HQGZWW does not have such effect.The protective role of HQGZWW against EAE might thus depend on the Th2 cytokine secretion profile induced by either MOG;specific CD8^(+)or CD4^(+)T-cells.
基金This study was supported by the Chinese National Natural Science Foundation(Grant Nos.31970771,82071396,81771345)the Shaanxi Provincial Key R&D Foundation(Grant Nos.2021ZDLSF03-09)+2 种基金the Science and Technology Projects of Ningxia Autonomous Region Key R&D Programs(2018BFG02017)the Natural Science Foundation of Ningxia Province,China(Grant Nos.2020AAC03397)the Fundamental Research Funds for the Central Universities(Grant Nos.GK202007022,GK202105002,TD2020039Y,2020CSLZ009,2021CSZL008).
文摘The lack of targeted and high-efficiency drug delivery to the central nervous system(CNS)nidus is the main problem in the treatment of demyelinating disease.Extracellular vesicles(EVs)possess great promise as a drug delivery vector given their advanced features.However,clinical applications are limited because of their inadequate targeting ability and the“dilution effects”after systemic administration.Neural stem cells(NSCs)supply a plentiful source of EVs on account of their extraordinary capacity for self-renewal.Here,we have developed a novel therapeutic system using EVs from modified NSCs with high expressed ligand PDGF-A(EVPs)and achieve local delivery.It has been demonstrated that EVPs greatly enhance the target capability on oligodendrocyte lineage.Moreover,EVPs are used for embedding triiodothyronine(T3),a thyroid hormone that is critical for oligodendrocyte development but has serious side effects when systemically administered.Our results demonstrated that systemic injection of EVPs+T3,versus EVPs or T3 administration individually,markedly alleviated disease development,enhanced oligodendrocyte survival,inhibited myelin damage,and promoted myelin regeneration in the lesions of experimental autoimmune encephalomyelitis mice.Taken together,our findings showed that engineered EVPs possess a remarkable CNS lesion targeting potential that offers a potent therapeutic strategy for CNS demyelinating diseases as well as neuroinflammation.
基金Work in the authors’lab was supported by grants from the NIH(AI139046 to L.V.K.and 1ZIAES10328601 to J.M.)the National Multiple Sclerosis Society(60006625 to L.V.K.)+3 种基金Core Services were performed through the Vanderbilt Digestive Disease Research Center(NIH grant P30DK058404)the Vanderbilt Ingram Cancer Center(NIH grant P30CA68485)the Vanderbilt Diabetes Research and Training Center(NIH grant P60DK020593)J.L.P.was supported by predoctoral NIH training grants(T32HL069765 and T32AR059039).
文摘The PIK3C3/VPS34 subunit of the class III phosphatidylinositol 3-kinase(PtdIns3K)complex plays a role in both canonical and noncanonical autophagy,key processes that control immune-cell responsiveness to a variety of stimuli.Our previous studies found that PIK3C3 is a critical regulator that controls the development,homeostasis,and function of dendritic and T cells.In this study,we investigated the role of PIK3C3 in myeloid cell biology using myeloid cell-specific Pik3c3-deficient mice.We found that Pik3c3-deficient macrophages express increased surface levels of major histocompatibility complex(MHC)class I and class II molecules.In addition,myeloid cell-specific Pik3c3 ablation in mice caused a partial impairment in the homeostatic maintenance of macrophages expressing the apoptotic cell uptake receptor TIM-4.Pik3c3 deficiency caused phenotypic changes in myeloid cells that were dependent on the early machinery(initiation/nucleation)of the classical autophagy pathway.Consequently,myeloid cell-specific Pik3c3-deficient animals showed significantly reduced severity of experimental autoimmune encephalomyelitis(EAE),a primarily CD4^(^(+))T-cell-mediated mouse model of multiple sclerosis(MS).This disease protection was associated with reduced accumulation of myelin-specific CD4^(^(+))T cells in the central nervous system and decreased myeloid cell IL-1βproduction.Further,administration of SAR405,a selective PIK3C3 inhibitor,delayed disease progression.Collectively,our studies establish PIK3C3 as an important regulator of macrophage functions and myeloid cell-mediated regulation of EAE.Our findings also have important implications for the development of small-molecule inhibitors of PIK3C3 as therapeutic modulators of MS and other autoimmune diseases.
文摘Objective To investigate the effets of nordihydroguaiaretic acid(NDGA)on the expression of IL-6,IL-17and TGF-βin mice with experimental autoimmune encephalomyelitis(EAE).Methods C57BL/6 mice were immunized with MOG35-55 to induce EAE.The 54 mice were randomly and equally divided into control group,model group and treatment group.Mice in treatment
基金supported by the National Natural Science Foundation of China(82074043,82104425,82374065,and 81673626)the China Postdoctoral Science Foundation(2021M702217).
文摘Early growth response protein 1(Egr-1)triggers the transcription of many genes involved in cell growth,differentiation,synaptic plasticity,and neurogenesis.However,its mechanism in neuronal survival and degeneration is still poorly understood.This study demonstrated that Egr-1 was down-regulated at mRNA and protein levels in the central nervous system(CNS)of experimental autoimmune encephalomyelitis(EAE)mice.Egr-1 knockout exacerbated EAE progression in mice,as shown by increased disease severity and incidence;it also aggravated neuronal apoptosis,which was associated with weakened activation of the BDNF/TGFβ1/MAPK/Akt signaling pathways in the CNS of EAE mice.Consistently,Egr-1 siRNA promoted apoptosis but mitigated the activation of BDNF/TGFβ1/MAPK/Akt signaling in SH-SY5Y cells.Our results revealed that Egr-1 is a crucial regulator of neuronal survival in EAE by regulating TGFβ1-mediated signaling activation,implicating the important role of Egr-1 in the pathogenesis of multiple sclerosis as a potential novel therapy target.
基金supported by a grant from the National Research Foundation(NRF)of Korea funded by the Korean Government,No.NRF-2022R1A2C1004022(to CM)。
文摘Multiple sclerosis is a chronic autoimmune disease of the central nervous system and is generally considered to be a non-traumatic,physically debilitating neurological disorder.In addition to experiencing motor disability,patients with multiple sclerosis also experience a variety of nonmotor symptoms,including cognitive deficits,anxiety,depression,sensory impairments,and pain.However,the pathogenesis and treatment of such non-motor symptoms in multiple scle rosis are still under research.Preclinical studies for multiple sclerosis benefit from the use of disease-appropriate animal models,including experimental autoimmune encephalomyelitis.Prior to understanding the pathophysiology and developing treatments for non-motor symptoms,it is critical to chara cterize the animal model in terms of its ability to replicate certain non-motor features of multiple sclerosis.As such,no single animal model can mimic the entire spectrum of symptoms.This review focuses on the non-motor symptoms that have been investigated in animal models of multiple sclerosis as well as possible underlying mechanisms.Further,we highlighted gaps in the literature to explain the nonmotor aspects of multiple sclerosis in expe rimental animal models,which will serve as the basis for future studies.
基金supported by the National Research Foundation (NRF)of Korea Grant funded by the Korean Government (NRF-2022R1A2C100402212RS-2023-00219517)。
文摘Structural plasticity is critical for the functional diversity of neurons in the brain.Experimental autoimmune encephalomyelitis(EAE)is the most commonly used model for multiple sclerosis(MS),successfully mimicking its key pathological features(inflammation,demyelination,axonal loss,and gliosis)and clinical symptoms(motor and non-motordysfunctions).Recentstudieshave demonstrated the importance of synaptic plasticity in EAE pathogenesis.In the present study,we investigated the features of behavioral alteration and hippocampal structural plasticity in EAE-affected mice in the early phase(11 days post-immunization,DPI)and chronic phase(28DPI).EAE-affected mice exhibited hippocampus-related behavioral dysfunction in the open field test during both early and chronic phases.Dendritic complexity was largely affected in the cornu ammonis 1(CA1)and CA3 apical and dentate gyrus(DG)subregions of the hippocampus during the chronic phase,while this effect was only noted in the CA1 apical subregion in the early phase.Moreover,dendritic spine density was reduced in the hippocampal CA1 and CA3 apical/basal and DG subregions in the early phase of EAE,but only reduced in the DG subregion during the chronic phase.Furthermore,mRNA levels of proinflammatory cytokines(Il1β,Tnfα,and Ifnγ)and glial cell markers(Gfap and Cd68)were significantly increased,whereas the expression of activity-regulated cytoskeletonassociated protein(ARC)was reduced during the chronic phase.Similarly,exposure to the aforementioned cytokines in primary cultures of hippocampal neurons reduced dendritic complexity and ARC expression.Primary cultures of hippocampal neurons also showed significantly reduced extracellular signal-regulated kinase(ERK)phosphorylation upon treatment with proinflammatory cytokines.Collectively,these results suggest that autoimmune neuroinflammation alters structural plasticity in the hippocampus,possibly through the ERK-ARC pathway,indicating that this alteration may be associated with hippocampal dysfunctions in EAE.
基金supported by Vice Chancellor for Research,Alborz University of Medical Sciences,No.1394-01-01-1050
文摘Hydroxycitric acid(HCA) is derived primarily from the Garcinia plant and is widely used for its anti-inflammatory effects. Multiple sclerosis can cause an inflammatory demyelination and axonal damage. In this study, to validate the hypothesis that HCA exhibits therapeutic effects on multiple sclerosis, we established female C57BL/6 mouse models of multiple sclerosis, i.e., experimental autoimmune encephalomyelitis,using Complete Freund's Adjuvant(CFA) emulsion containing myelin oligodendrocyte glycoprotein(35–55). Treatment with HCA at 2 g/kg/d for 3 weeks obviously improved the symptoms of nerve injury of experimental autoimmune encephalomyelitis mice, decreased serum interleulin-6, tumor necrosis factor alpha, nitric oxide, and malondialdehyde levels, and increased superoxide dismutase and glutathione reductase activities. These findings suggest that HCA exhibits neuroprotective effects on multiple sclerosis-caused nerve injury through ameliorating inflammation and oxidative stress.
