BACKGROUND:Olfactory ensheathing cell transplantation can activate axonal regeneration and enhance myelin repair,which are beneficial for treating demyelinating diseases. OBJECTIVE:To explore the effects of olfactory ...BACKGROUND:Olfactory ensheathing cell transplantation can activate axonal regeneration and enhance myelin repair,which are beneficial for treating demyelinating diseases. OBJECTIVE:To explore the effects of olfactory ensheathing cell transplantation on myelin repair, synaptophysin expression,and motor function in a rat model of experimental allergic encephalomyelitis. DESIGN,TIME AND SETTING:A randomized,controlled experiment was performed at the Laboratory of Provincial Hospital affiliated to Shandong University between August 2006 and September 2007. MATERIALS:Dibenzylamine(Hoechst 33342),luxol fast blue,and rabbit anti-rat synaptophysin antibody were provided by Sigma,USA. METHODS:Olfactory ensheathing cells extracted from neonatal Wistar rats were cultured for 10-14 days and labeled with dibenzylamine.Spinal cord extracted from a healthy guinea pig was homogenized and equally mixed with complete Freund's adjuvant;thereafter,the mixture was intracutaneously injected into two posterior voix pedis of healthy male Wistar rats to establish models of experimental allergic encephalomyelitis.Rats were randomly divided into a control encephalomyelitis group and an olfactory ensheathing cell transplantation group,36 rats in each group.Physiological saline(2μL) or an olfactory ensheathing cell suspension(2μL) was separately injected along lateral cerebral ventricle at day 7 post-model induction. MAIN OUTCOME MEASURES:The migration and distribution of olfactory ensheathing cells were observed under fluorescence microscopy;myelin repair was detected using hematoxylin-eosin staining and luxol fast blue staining;synaptophysin expression was measured using immunohistochemical staining;motor function was evaluated using a motor function scale. RESULTS:Olfactory ensheathing cells could survive in vivo and migrate to the distal end of the transplant focus and spinal cord,and survived 21 days.Hematoxylin-eosin staining and luxol fast blue staining indicated that myelin in the transplantation group was intact,and the inflammatory focus gradually disappeared.Transplantation increased synaptophysin expression(P<0.05 versus control) and motor function(P<0.05). CONCLUSION:Olfactory ensheathing cell transplantation can promote myelin repair,increase synaptophysin protein expression,and ameliorate motor function in a rat model of experimental allergic encephalomyelitis.展开更多
Olfactory ensheathing cells(OECs) can promote axonal regeneration and remyelination for the treatment of spinal cord injury.OECs can also treat experimental allergic encephalomyelitis(EAE),but it remains unclear wheth...Olfactory ensheathing cells(OECs) can promote axonal regeneration and remyelination for the treatment of spinal cord injury.OECs can also treat experimental allergic encephalomyelitis(EAE),but it remains unclear whether OECs might be rejected by the immune system in the brain including the destruction of the blood-brain barrier under inflammation,the release of inflammatory factors,the activation of local antigen-presenting cells(e.g.,microglia cells) and antigen drainage.We found that OECs expressed major histocompatibility complex(MHC)-I molecules on the cell surface,barely expressed MHC-Ⅱ,but MHC-Ⅱ could be induced by interferon-γ,suggesting that OECs have certain immunogenicity.When OECs were transplanted into normal animal brains,no OECs were phagocytosed by dendritic cells in the cervical lymph node,and OECs did not induce lymphocyte proliferation,which indicates that OECs share some immune privilege under normal conditions.However,OECs in the rat EAE brain were phagocytosed by dendritic cells in the cervical lymph node and enhanced lymphocyte proliferation.These findings suggest that OECs are rejected because of increased immunogenicity in EAE brain,and that brain inflammation,in particular activated dendritic cells,may be a prerequisite for rejecting OECs.展开更多
Experimental allergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis.Th1 cells play an important role in the pathogenesis of experimental allergic encephalomyeli...Experimental allergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis.Th1 cells play an important role in the pathogenesis of experimental allergic encephalomyelitis.This study determined the potential effect of programmed cell death 1ligand 1 in the pathogenesis of experimental allergic encephalomyelitis induced by injecting myelin oligodendrocyte glycoprotein,complete Freund’s adjuvant and Bordetella pertussis toxin into C57BL/6J mice.Experimental allergic encephalomyelitis mice developed disease and showed inflammatory changes in the central nervous system by hematoxylin-eosin staining of spinal cord pathological sections,demyelination by Luxol fast-blue staining and clinical manifestations.The expression of programmed cell death 1 ligand 1 in mice was detected by immunohistochemistry,flow cytometry and western blot analysis.The expression of programmed cell death 1 ligand 1 in the spinal cord and splenocytes of mice was significantly increased compared with normal mice.Our findings suggest the involvement of programmed cell death 1 ligand 1 in the pathogenesis of experimental allergic encephalomyelitis and suggest this should be studied in multiple sclerosis.展开更多
BACKGROUND: Neuropeptide Y (NPY) may influence differentiation of Th cells. It is assumed that the immunological pathology of experimental allergic encephalomyelitis (EAE) is related to abnormal differentiation of Th ...BACKGROUND: Neuropeptide Y (NPY) may influence differentiation of Th cells. It is assumed that the immunological pathology of experimental allergic encephalomyelitis (EAE) is related to abnormal differentiation of Th cells OBJECTIVE: To investigate the effect of NPY on white matter demyelination, the serum levels interleukin-4 (IL-4) and gamma-interferon (IFN-γ), as well as EAE pathogenesis in an EAE guinea pig model following NPY injection into the lateral cerebral ventricle. DESIGN, TIME AND SETTING: A randomized controlled animal study, which was performed in the Infection Immunity Animal Laboratory, Affiliated Hospital of Luzhou Medical College, China, from October 2005 to April 2006. MATERIALS: Thirty healthy female guinea pigs of 8–12 weeks of age, and 10 healthy female rats of three months of age were used. NPY was provided by Sigma Company, USA. NPY kit was provided by Beijing Huaying Biotechnology Institute, China. METHODS: Thirty guinea pigs were randomly divided into three groups: normal control group, EAE model group, and NPY intervention group (n =10 per group). Normal control group and EAE model group: Saline (10 μL, once) was injected into the lateral cerebral ventricle. After one week, the same volume of Freund’s adjuvant complete was either injected subcutaneously into two post-palms or EAE was modeled. NPY intervention group: EAE was modeled after one week and NPY was injected (10 μL of 6 nmol NPY, once) into the lateral cerebral ventricle. Myelin basic protein (MBP) antigen made from rat spinal cord homogenate and Freund’s adjuvant complete were injected subcutaneously into both post-palms (0.2 mL per palm) to establish the EAE model. MAIN OUTCOME MEASURES: White matter demyelination of the cerebrum, cerebellum, brain stem, and spinal cord were observed by light microscopy after HE staining. Levels of serum IFN-γ and IL-4 were detected by the double antibody sandwich ABC-ELISA technique. NPY content was detected by radioimmunoassay. RESULTS: Pathological alterations in the NPY intervention groups were reduced compared to those in the EAE model group, suggesting a reduction and remission of white matter demyelination with NPY treatment. When compared to the model group, the serum IL-4 level was increased in the NPY intervention group during the high-frequent EAE stage (P < 0.01), but the serum IFN-γ level was decreased (P < 0.01). Furthermore, the EAE latency was prolonged (P < 0.01), the neurological scores were decreased in the high-frequent EAE stage (P < 0.01), and the death rate was decreased (P < 0.05). NPY content and the serum IL-4 level at the peak stage were positively correlated with those in the latent phase (r =0.863-0.900, P < 0.01), but negatively correlated with neurological scores at the peak stage (r= -0.068 to –0.863, P < 0.05–0.01). The IFN-γ level at the peak stage was negatively correlated to that in the latent phase (r = -0.683–0.650, P < 0.05), but positively correlated to neurological scores at the peak stage (r =0.975, 0.845, P < 0.05). CONCLUSION: NPY injection into the lateral cerebral ventricle can promote the secretion of IL-4, inhibit the production of IFN-γ, relieve white matter demyelination, and inhibit EAE attack in an experimental model of EAE.展开更多
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
To detect the function of proteolipid protein (PLP) peptide (residues 56-70) sp ecific CD 4 + T cells in experimental allergic encephalomyelitis (EAE) in Bioz zi AB/H mice (H 2A g7 ) Methods Biozzi AB/H mice were immu...To detect the function of proteolipid protein (PLP) peptide (residues 56-70) sp ecific CD 4 + T cells in experimental allergic encephalomyelitis (EAE) in Bioz zi AB/H mice (H 2A g7 ) Methods Biozzi AB/H mice were immunized by synthetic PLP 56 70 peptide (DYEYLINVIH AFQYV) which was emulsified by sonication with complete Freund’s adjuvant, a EAE model proven histologically and clinically Murine splenocytes and spinal cord infiltrated (SCI) T cells were stimulated by PLP 56 70 , then the CD 4 + T cells were isolated by Dynabeads, and confirmed by staining with anti CD 4 antibody Finally, the IL2 bioassay and IFN γ/IL4 ELISA were done to detect T cell proliferation and cytokine secretion after PLP 56 70 stimulation Results The histology of murine spinal cord showed a great number of lymphocytes infiltr ated the spinal cord; the clinical signs showed high scores (4 3) on the peak, as well as a good EAE model After being isolated by Dynabea ds , CD 4 + T cells showed high purification (>99%) by staining with anti CD 4 antibody IL2 bioassay showed that those T cells were PLP 56 70 specific T cells ELISA showed that those T cells had high IFN γ/IL4 ratio , indicating that they are T helper 1 (Th1) cells Conclusions PLP 56 70 specific splenocytes and SCI CD 4 + T cells in EAE from Bioz zi AB/H mice were detected and showed that both of them were P LP 56 70 specific Th1 cells It is beneficial to understand what kind o f role these T cells play in the development of展开更多
BACKGROUND:Studies have demonstrated that experimental autoimmune encephalomyelitis(EAE) onset correlates with increased interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α) expression.Oxymatrine(OM) has been sh...BACKGROUND:Studies have demonstrated that experimental autoimmune encephalomyelitis(EAE) onset correlates with increased interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α) expression.Oxymatrine(OM) has been shown to inhibit autoimmune responses,but there are no reports showing that it could prevent the development of EAE.OBJECTIVE:To observe the effect of OM on serum levels of IFN-γ and TNF-α in a rat model of EAE.DESIGN,TIME AND SETTING:A randomized,controlled,animal study was performed at the Experimental Animal Center of Henan Academy of Chinese Medicine and at the Key Disciplines Laboratory Clinical Medicine of Henan Province between July and December 2008.MATERIALS:OM was purchased from Chia-tai Tianqing Pharmaceutical,China;complete Freund's adjuvant was purchased from Sigma,USA.METHODS:Forty female Wistar rats were randomly assigned to four groups:EAE model(M),low-dose OM treatment(OM-L),high-dose OM treatment(OM-H),and normal control(N,no immunization),with 10 rats in each group.EAE was established in the M,OM-L,and OM-H groups following immunization with Guinea pig spinal cord homogenate and complete Freund's adjuvant.The M and N groups were intraperitoneally injected with normal saline(6.7 mL/kg per day),the OM-L group received an intraperitoneal injection of OM(100 mg/kg per day),and the OM-H group received OM(150 mg/kg per day).MAIN OUTCOME MEASURES:At 16 days after immunization,the degree of histopathological changes in the spinal cord was assessed by hematoxylin-eosin stanining.Enzyme-linked immunosorbent assay was used to detect serum levels of IFN-γ,and radioimmunoassay was utilized to determine serum TNF-α level.Neurological scores were measured on a daily basis according to a 0-5 scale.RESULTS:Daily injections of OM,both high and low doses,resulted in decreased neurological scores in EAE rats(P < 0.01),as well as reduced cellular infiltration in the spinal cord and decreased levels of serum IFN-γ and TNF-α(P < 0.01).CONCLUSION:OM reduced the onset and severity of EAE,which correlated with decreased IFN-γ and TNF-α expression.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
AIM: To assess the efficacy of topical Semaphorin 3A(SEMA3A) in the treatment of allergic conjunctivitis.METHODS: Experimental allergic conjunctivitis(EAC)mice model induced by short ragweed pollen(SRW) in 4-week-old ...AIM: To assess the efficacy of topical Semaphorin 3A(SEMA3A) in the treatment of allergic conjunctivitis.METHODS: Experimental allergic conjunctivitis(EAC)mice model induced by short ragweed pollen(SRW) in 4-week-old of BALB/c mice, mice were evaluated using haematoxylin and eosin(H&E) staining, immunofluor-escence and light microscope photographs. Early phase took the samples in 24 h after instillation and late phase took the samples between 4 to 14 d after the start of treatment. The study use of topical SEMA3A(10 U, 100 U,1000 U) eye drops and subconjunctival injection of SEMA3 A with same concentration. For comparison, five types of allergy eyedrops were quantified using clinical characteristics.· RESULTS: Clinical score of composite ocular symptoms of the mice treated with SEMA3 A were significantly decreased both in the immediate phase and the late phase compared to those treated with commercial ophthalmic formulations and non-treatment mice. SEMA3 A treatment attenuates infiltration of eosinophils entering into conjunctiva in EAC mice. The score of eosinophil infiltration in the conjunctiva of SEMA3 A 1000 U-treated group were significantly lower than low-concentration of SEMA3 A treated groups and non-treated group. SEMA3 A treatment also suppressed T-cell proliferation in vitro and decreased serum total Ig E levels in EAC mice. Moreover, treatment of SEMA3 A suppressed Th2-related cytokines(IL-5, IL-13 and IL-4)and pro-inflammatory cytokines(IFN-γ, IL-17 and TNF-α)release, but increased regulatory cytokine IL-10 concentration in the conjunctiva of EAC mice.CONCLUSION: SEMA3 A as a biological agent, showed the beneficial activity in ocular allergic processes with the less damage to the intraocular tissue. It is expected that SEMA3 A may be contributed in patients with a more severe spectrum of refractory ocular allergic diseases including allergic conjunctivitis in the near future.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
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
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
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
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
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
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
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
基金Supported by:the National Natural Science Foundation of China,No.30770751the Foundation for Youth of Shandong Bureau of Public Health,No.2007QZ002the Doctoral Foundation of Shandong Scientific and Technological Bureau,No. 2008BS03004
文摘BACKGROUND:Olfactory ensheathing cell transplantation can activate axonal regeneration and enhance myelin repair,which are beneficial for treating demyelinating diseases. OBJECTIVE:To explore the effects of olfactory ensheathing cell transplantation on myelin repair, synaptophysin expression,and motor function in a rat model of experimental allergic encephalomyelitis. DESIGN,TIME AND SETTING:A randomized,controlled experiment was performed at the Laboratory of Provincial Hospital affiliated to Shandong University between August 2006 and September 2007. MATERIALS:Dibenzylamine(Hoechst 33342),luxol fast blue,and rabbit anti-rat synaptophysin antibody were provided by Sigma,USA. METHODS:Olfactory ensheathing cells extracted from neonatal Wistar rats were cultured for 10-14 days and labeled with dibenzylamine.Spinal cord extracted from a healthy guinea pig was homogenized and equally mixed with complete Freund's adjuvant;thereafter,the mixture was intracutaneously injected into two posterior voix pedis of healthy male Wistar rats to establish models of experimental allergic encephalomyelitis.Rats were randomly divided into a control encephalomyelitis group and an olfactory ensheathing cell transplantation group,36 rats in each group.Physiological saline(2μL) or an olfactory ensheathing cell suspension(2μL) was separately injected along lateral cerebral ventricle at day 7 post-model induction. MAIN OUTCOME MEASURES:The migration and distribution of olfactory ensheathing cells were observed under fluorescence microscopy;myelin repair was detected using hematoxylin-eosin staining and luxol fast blue staining;synaptophysin expression was measured using immunohistochemical staining;motor function was evaluated using a motor function scale. RESULTS:Olfactory ensheathing cells could survive in vivo and migrate to the distal end of the transplant focus and spinal cord,and survived 21 days.Hematoxylin-eosin staining and luxol fast blue staining indicated that myelin in the transplantation group was intact,and the inflammatory focus gradually disappeared.Transplantation increased synaptophysin expression(P<0.05 versus control) and motor function(P<0.05). CONCLUSION:Olfactory ensheathing cell transplantation can promote myelin repair,increase synaptophysin protein expression,and ameliorate motor function in a rat model of experimental allergic encephalomyelitis.
基金Young Talent Innovation Program of Fujian Province Department of Science and Technology,No.2006F3032
文摘Olfactory ensheathing cells(OECs) can promote axonal regeneration and remyelination for the treatment of spinal cord injury.OECs can also treat experimental allergic encephalomyelitis(EAE),but it remains unclear whether OECs might be rejected by the immune system in the brain including the destruction of the blood-brain barrier under inflammation,the release of inflammatory factors,the activation of local antigen-presenting cells(e.g.,microglia cells) and antigen drainage.We found that OECs expressed major histocompatibility complex(MHC)-I molecules on the cell surface,barely expressed MHC-Ⅱ,but MHC-Ⅱ could be induced by interferon-γ,suggesting that OECs have certain immunogenicity.When OECs were transplanted into normal animal brains,no OECs were phagocytosed by dendritic cells in the cervical lymph node,and OECs did not induce lymphocyte proliferation,which indicates that OECs share some immune privilege under normal conditions.However,OECs in the rat EAE brain were phagocytosed by dendritic cells in the cervical lymph node and enhanced lymphocyte proliferation.These findings suggest that OECs are rejected because of increased immunogenicity in EAE brain,and that brain inflammation,in particular activated dendritic cells,may be a prerequisite for rejecting OECs.
基金financially sponsored by the Natural Science Foundation of Jiangsu Province in China,(General Program),No.BK2011267
文摘Experimental allergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis.Th1 cells play an important role in the pathogenesis of experimental allergic encephalomyelitis.This study determined the potential effect of programmed cell death 1ligand 1 in the pathogenesis of experimental allergic encephalomyelitis induced by injecting myelin oligodendrocyte glycoprotein,complete Freund’s adjuvant and Bordetella pertussis toxin into C57BL/6J mice.Experimental allergic encephalomyelitis mice developed disease and showed inflammatory changes in the central nervous system by hematoxylin-eosin staining of spinal cord pathological sections,demyelination by Luxol fast-blue staining and clinical manifestations.The expression of programmed cell death 1 ligand 1 in mice was detected by immunohistochemistry,flow cytometry and western blot analysis.The expression of programmed cell death 1 ligand 1 in the spinal cord and splenocytes of mice was significantly increased compared with normal mice.Our findings suggest the involvement of programmed cell death 1 ligand 1 in the pathogenesis of experimental allergic encephalomyelitis and suggest this should be studied in multiple sclerosis.
文摘BACKGROUND: Neuropeptide Y (NPY) may influence differentiation of Th cells. It is assumed that the immunological pathology of experimental allergic encephalomyelitis (EAE) is related to abnormal differentiation of Th cells OBJECTIVE: To investigate the effect of NPY on white matter demyelination, the serum levels interleukin-4 (IL-4) and gamma-interferon (IFN-γ), as well as EAE pathogenesis in an EAE guinea pig model following NPY injection into the lateral cerebral ventricle. DESIGN, TIME AND SETTING: A randomized controlled animal study, which was performed in the Infection Immunity Animal Laboratory, Affiliated Hospital of Luzhou Medical College, China, from October 2005 to April 2006. MATERIALS: Thirty healthy female guinea pigs of 8–12 weeks of age, and 10 healthy female rats of three months of age were used. NPY was provided by Sigma Company, USA. NPY kit was provided by Beijing Huaying Biotechnology Institute, China. METHODS: Thirty guinea pigs were randomly divided into three groups: normal control group, EAE model group, and NPY intervention group (n =10 per group). Normal control group and EAE model group: Saline (10 μL, once) was injected into the lateral cerebral ventricle. After one week, the same volume of Freund’s adjuvant complete was either injected subcutaneously into two post-palms or EAE was modeled. NPY intervention group: EAE was modeled after one week and NPY was injected (10 μL of 6 nmol NPY, once) into the lateral cerebral ventricle. Myelin basic protein (MBP) antigen made from rat spinal cord homogenate and Freund’s adjuvant complete were injected subcutaneously into both post-palms (0.2 mL per palm) to establish the EAE model. MAIN OUTCOME MEASURES: White matter demyelination of the cerebrum, cerebellum, brain stem, and spinal cord were observed by light microscopy after HE staining. Levels of serum IFN-γ and IL-4 were detected by the double antibody sandwich ABC-ELISA technique. NPY content was detected by radioimmunoassay. RESULTS: Pathological alterations in the NPY intervention groups were reduced compared to those in the EAE model group, suggesting a reduction and remission of white matter demyelination with NPY treatment. When compared to the model group, the serum IL-4 level was increased in the NPY intervention group during the high-frequent EAE stage (P < 0.01), but the serum IFN-γ level was decreased (P < 0.01). Furthermore, the EAE latency was prolonged (P < 0.01), the neurological scores were decreased in the high-frequent EAE stage (P < 0.01), and the death rate was decreased (P < 0.05). NPY content and the serum IL-4 level at the peak stage were positively correlated with those in the latent phase (r =0.863-0.900, P < 0.01), but negatively correlated with neurological scores at the peak stage (r= -0.068 to –0.863, P < 0.05–0.01). The IFN-γ level at the peak stage was negatively correlated to that in the latent phase (r = -0.683–0.650, P < 0.05), but positively correlated to neurological scores at the peak stage (r =0.975, 0.845, P < 0.05). CONCLUSION: NPY injection into the lateral cerebral ventricle can promote the secretion of IL-4, inhibit the production of IFN-γ, relieve white matter demyelination, and inhibit EAE attack in an experimental model of EAE.
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To detect the function of proteolipid protein (PLP) peptide (residues 56-70) sp ecific CD 4 + T cells in experimental allergic encephalomyelitis (EAE) in Bioz zi AB/H mice (H 2A g7 ) Methods Biozzi AB/H mice were immunized by synthetic PLP 56 70 peptide (DYEYLINVIH AFQYV) which was emulsified by sonication with complete Freund’s adjuvant, a EAE model proven histologically and clinically Murine splenocytes and spinal cord infiltrated (SCI) T cells were stimulated by PLP 56 70 , then the CD 4 + T cells were isolated by Dynabeads, and confirmed by staining with anti CD 4 antibody Finally, the IL2 bioassay and IFN γ/IL4 ELISA were done to detect T cell proliferation and cytokine secretion after PLP 56 70 stimulation Results The histology of murine spinal cord showed a great number of lymphocytes infiltr ated the spinal cord; the clinical signs showed high scores (4 3) on the peak, as well as a good EAE model After being isolated by Dynabea ds , CD 4 + T cells showed high purification (>99%) by staining with anti CD 4 antibody IL2 bioassay showed that those T cells were PLP 56 70 specific T cells ELISA showed that those T cells had high IFN γ/IL4 ratio , indicating that they are T helper 1 (Th1) cells Conclusions PLP 56 70 specific splenocytes and SCI CD 4 + T cells in EAE from Bioz zi AB/H mice were detected and showed that both of them were P LP 56 70 specific Th1 cells It is beneficial to understand what kind o f role these T cells play in the development of
基金a Grant from the Natural Scientific Research Project of the Education Department of Henan Province,No. 2009A350009
文摘BACKGROUND:Studies have demonstrated that experimental autoimmune encephalomyelitis(EAE) onset correlates with increased interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α) expression.Oxymatrine(OM) has been shown to inhibit autoimmune responses,but there are no reports showing that it could prevent the development of EAE.OBJECTIVE:To observe the effect of OM on serum levels of IFN-γ and TNF-α in a rat model of EAE.DESIGN,TIME AND SETTING:A randomized,controlled,animal study was performed at the Experimental Animal Center of Henan Academy of Chinese Medicine and at the Key Disciplines Laboratory Clinical Medicine of Henan Province between July and December 2008.MATERIALS:OM was purchased from Chia-tai Tianqing Pharmaceutical,China;complete Freund's adjuvant was purchased from Sigma,USA.METHODS:Forty female Wistar rats were randomly assigned to four groups:EAE model(M),low-dose OM treatment(OM-L),high-dose OM treatment(OM-H),and normal control(N,no immunization),with 10 rats in each group.EAE was established in the M,OM-L,and OM-H groups following immunization with Guinea pig spinal cord homogenate and complete Freund's adjuvant.The M and N groups were intraperitoneally injected with normal saline(6.7 mL/kg per day),the OM-L group received an intraperitoneal injection of OM(100 mg/kg per day),and the OM-H group received OM(150 mg/kg per day).MAIN OUTCOME MEASURES:At 16 days after immunization,the degree of histopathological changes in the spinal cord was assessed by hematoxylin-eosin stanining.Enzyme-linked immunosorbent assay was used to detect serum levels of IFN-γ,and radioimmunoassay was utilized to determine serum TNF-α level.Neurological scores were measured on a daily basis according to a 0-5 scale.RESULTS:Daily injections of OM,both high and low doses,resulted in decreased neurological scores in EAE rats(P < 0.01),as well as reduced cellular infiltration in the spinal cord and decreased levels of serum IFN-γ and TNF-α(P < 0.01).CONCLUSION:OM reduced the onset and severity of EAE,which correlated with decreased IFN-γ and TNF-α expression.
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A grant-in-aid for Scientific Research from the Japanese Ministry of Education,Culture,Sports,Science and Technology
文摘AIM: To assess the efficacy of topical Semaphorin 3A(SEMA3A) in the treatment of allergic conjunctivitis.METHODS: Experimental allergic conjunctivitis(EAC)mice model induced by short ragweed pollen(SRW) in 4-week-old of BALB/c mice, mice were evaluated using haematoxylin and eosin(H&E) staining, immunofluor-escence and light microscope photographs. Early phase took the samples in 24 h after instillation and late phase took the samples between 4 to 14 d after the start of treatment. The study use of topical SEMA3A(10 U, 100 U,1000 U) eye drops and subconjunctival injection of SEMA3 A with same concentration. For comparison, five types of allergy eyedrops were quantified using clinical characteristics.· RESULTS: Clinical score of composite ocular symptoms of the mice treated with SEMA3 A were significantly decreased both in the immediate phase and the late phase compared to those treated with commercial ophthalmic formulations and non-treatment mice. SEMA3 A treatment attenuates infiltration of eosinophils entering into conjunctiva in EAC mice. The score of eosinophil infiltration in the conjunctiva of SEMA3 A 1000 U-treated group were significantly lower than low-concentration of SEMA3 A treated groups and non-treated group. SEMA3 A treatment also suppressed T-cell proliferation in vitro and decreased serum total Ig E levels in EAC mice. Moreover, treatment of SEMA3 A suppressed Th2-related cytokines(IL-5, IL-13 and IL-4)and pro-inflammatory cytokines(IFN-γ, IL-17 and TNF-α)release, but increased regulatory cytokine IL-10 concentration in the conjunctiva of EAC mice.CONCLUSION: SEMA3 A as a biological agent, showed the beneficial activity in ocular allergic processes with the less damage to the intraocular tissue. It is expected that SEMA3 A may be contributed in patients with a more severe spectrum of refractory ocular allergic diseases including allergic conjunctivitis in the near future.
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.
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 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.
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 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.
