Y-39983在实验性自身免疫性脑脊髓炎中的作用研究

目的 探讨Rho-ROCK信号途径与实验性自身免疫性脑脊髓炎(EAE)的关系及Y-39983在EAE中的作用.方法 动物被分为对照组(健康Lewis鼠)、EAE模型组及干预组,每组各30只.EAE模型组及干预组建立EAE动物模型.干预组于Lewis鼠发病第1天起连续3d给予腹腔注射Y-39983(20 μg/d),模型组同样于Lewis鼠发病第1天起连续3d给予腹腔注射生理盐水(20 μg/d).比较各组平均临床评分、发病高峰期持续时间及复发率;采用电镜观察法、HE染色、搔洛花青髓鞘染色、APC免疫荧光染色法观察各组脑组织病理变化;采用蛋白免疫印迹法检测各组蛋白脂蛋白(PLP)、髓磷脂碱性蛋白(MBP)及3-环腺苷酸-3-磷酸二酯酶(CNPase)等髓鞘蛋白分子及肌球蛋白轻链磷酸酶(p-MLC)、脑衰反应调节蛋白(CRMP-2)及LIM激酶2(LIMK2)等Rho激酶(ROCK)底物蛋白含量的变化.结果 干预组平均临床评分(1.076±0.682)、发病高峰期持续时间[(3.7±1.35)d]、疾病复发率(37.5％)均低于EAE模型组[分别为4.229±1.017、(5.3±1.46)d、46.67％](P＜0.05).干预组中脑组织中炎性反应及髓鞘脱失现象均较EAE模型组改善;脑组织中ROCK底物蛋白p-MLC、CRMP-2及LIMK2含量干预组较EAE模型组降低,髓鞘相关蛋白PLP、MBP及CNPase含量干预组较EAE模型组升高(均P＜0.05).结论 Y-39983可改善EAE临床症状,其可能作用机制为抑制ROCK激活,减少破坏轴突生长的底物蛋白生成,促进髓鞘蛋白形成及轴突再生.     

雄激素在实验性自身免疫性脑脊髓炎中的保护作用

多发性硬化(multiple sclerosis, MS)及其动物模型即实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)都具有性别差异,但其确切机制仍不清楚.近年来,大量研究发现雄激素与EAE的发病和病程有一定关系.此文就EAE存在的性别差异及雄激素在其中的作用做一综述.     

实验性变态反应性脑脊髓炎模型研究进展

实验性变态反应性脑脊髓炎是人类多发性硬化的理想动物模型,在临床神经免疫学研究中具有重要意义。本文主要介绍了实验性变态反应性脑脊髓炎模型制备的有关问题,包括敏感动物品系、性别、年龄的选择,抗原的种类及选择,免疫方法,模型的制备及病情评分等。     

实验性变态反应性脑脊髓炎与脑屏障变化

脑屏障改变是实验性变态反应性脑脊髓炎 (EAE)较早期的病理改变 ,它促使免疫细胞及免疫信息分子大量入脑 ,从而引起脑组织的免疫炎性反应。本文对脑屏障概念 ,EAE时脑屏障变化情况及其可能的机理作一综述。     

雌激素在实验性自身免疫性脑脊髓炎中的保护作用

多发性硬化(multiple sclerosis,MS)与其他多种自身免疫性疾病一样存在着性别差异,表现为:与男性相比,女性发病率较高,且初发时间也较早。女性患者在妊娠期伴随着体内雌激素水平的波动,MS患者复发次数和严重程度也出现明显变化[1]。在MS动物模型即实验性自身免疫性脑脊髓炎(exp     

实验性变态反应性脑脊髓炎的病理研究

目的　研究实验性变态反应性脑脊髓炎 (EAE)的病理变化。方法　利用光镜、电镜观察豚鼠 EAE模型的组织学改变。结果　光镜下可见脑实质及珠网膜下腔小血管充血 ,血管周围炎细胞浸润 ,呈“袖套”状改变 ,侧脑室旁白质及脊髓后联合、侧角出现髓鞘肿胀、崩解或消失。电镜下可见血管周围明显脱髓鞘 ,髓鞘呈松散状 ,部分区域斑块状脱落 ,明暗相间结构消失 ,轴突细胞器消失 ,神经元细胞尼 (氏 )小体消失 ,内质网扩张脱颗粒。结论　 EAE的病理改变主要为血管周围炎性浸润及白质脱髓鞘。     

雄激素在实验性自身免疫性脑脊髓炎中的保护作用（综述）

多发性硬化(multiple sclerosis，MS)及其动物模型即实验性自身免疫性脑脊髓炎(experiment alautimmune encephalomyelitis，EAE)都具有性别差异，但其确切机制仍不清楚。近年来，大量研究发现雄激素与EAE的发病和病程有一定关系。此文就EAE存在的性别差异及雄激素在其中的作用做一综述。     

小鼠实验性自身免疫性脑脊髓炎的病理变化

目的用髓鞘少突胶质细胞糖蛋白多肽(MOG35-55)诱发实验性自身免疫性脑脊髓炎(experi-m ental autoimmune encephalomyelitis,EAE)小鼠模型。方法应用MOG35-55抗原加完全弗氏佐剂免疫C57BL/6小鼠,利用光镜、电镜观察小鼠组织学改变。结果光镜下可见小血管周围炎细胞浸润,呈袖套状改变、血管周围明显脱髓鞘及神经元变性,B ieschowsky银染显示大量轴索肿胀和轴索卵形体的形成,电镜下可见髓鞘结构松散、断裂或融合,包括不同程度的髓鞘重建,脊髓病变广泛,程度重于脑部。结论EAE的病理改变为血管周围炎性细胞浸润、白质脱髓鞘及髓鞘重建。     

实验性变态反应性脑脊髓炎与细胞凋亡

实验性变态反应性脑脊髓炎 (EAE)是研究人类多发性硬化 (MS)的经典动物模型 ,EAE发病和愈复过程中细胞凋亡的情况也是目前神经生物学的研究热点。研究者们发现 ,EAE时CNS损伤区有自身反应性T淋巴细胞的凋亡 ,而且这种细胞凋亡促进了EAE的愈复和耐受状态的形成 ,这就为EAE乃至人类MS的防治提供了一条新思路。有关EAE其他细胞凋亡和影响细胞凋亡的因素也有一些研究。本综述对近年来这些方面的研究进展做了较为全面的回顾     

实验性变态反应性脑脊髓炎几种细胞因子的检测

目的和方法　为研究细胞因子在实验性变态反应性脑脊髓炎 (EAE)及多发性硬化 (MS)发病机制中的作用 ,用 CTL L细胞株滴定法检测了 EAE动物脾细胞培养上清中白细胞介素 - 2 (IL- 2 )的生物学活性 ,用EL ISA分别检测了白细胞介素 - 4 (IL - 4 )及干扰素 -γ(IFN-γ)水平。结果　 EAE组 IL - 2活性及 IFN-γ水平均显著高于正常对照组 (均为 P<0 .0 5) ,而其 IL- 4水平显著低于对照组 (P<0 .0 1)。结论　 IL- 2、IFN- γ等细胞因子参与了 EAE的发病 ,并对 IL- 4的产生有抑制作用。     

Chronic progressive experimental allergic encephalomyelitis (EAE) in adult guinea pigs

Adult Hartley guinea pigs were inoculated with an increased dose of whole central nervous system tissue in complete Freund's adjuvant in order to determine the effects of high dose of neuroantigens on the clinical and pathological course of experimental allergic encephalomyelitis (EAE). An increased survival rate during the first attack and a change in the course of the disease from an acute fatal one to chronic progressive EAE were observed. The main clinicopathological features of the disease included a delayed onset of mild neurological signs and the presence of large demyelinated plaques in the brain and spinal cord. The relationship of the chronic progressive course of the disease and the induction of high-zone tolerance is discussed.     

甲基泼尼松龙对实验性变态反应性脑脊髓炎大鼠脑组织 Nogo-A 及其受体 NgR 蛋白表达的影响

目的 探讨甲基泼尼松龙(MP)对实验性变态反应性脑脊髓炎(EAE)大鼠脑组织Nogo-A及其受体(NgR)蛋白表达的影响.方法 将60只Wistar大鼠随机分为正常对照组(10只)和实验组(50只).实验组大鼠通过皮下注射粗制髓磷脂碱性蛋白建立EAE模型,24只造模成功大鼠随机分为EAE组、大剂量MP(MP-H)组和小...     

On the role of peripheral macrophages during active experimental allergic encephalomyelitis (EAE)

Experimental allergic encephalitis (EAE) is an experimental autoimmune inflammatory condition of the central nervous system (CNS) that serves as a disease model for multiple sclerosis (MS). The primary effector mechanisms of the immune system leading to tissue destruction during EAE remain still controversial. T-cells, microglia, and macrophages infiltrating the brain parenchyma are suggested to be involved. To clarify the role of these cells during disease Lewis rats were immunised with different immunisation protocols: Immunisation with myelin basic protein (MBP) in complete Freunds adjuvant (CFA) containing high dose of mycobacterial components induced severe disease, whereas immunisation with low dose of mycobacterial components induced only mild disease. Severely and mildly diseased animals were analysed with respect to infiltration of T-cells, macrophages and upregulation of MHC class II molecules on microglia in the brain. All immunised rats showed high T-cell infiltration accompanied by microglia activation. The degree of disease and the infiltration of macrophages varied with dose of adjuvant. Lowering the dose of adjuvant prevented the development of disease but also the influx of peripheral macrophages into the brain without affecting the peripheral T-cell response to the autoantigen. Thus, appearance of (autoreactive) T-cells in the brain and microglia activation were probably not sufficient for development of disease. It can be concluded that peripheral macrophages play an essential or even key role in the pathogenesis of active EAE.     

实验性自身免疫性脑脊髓炎大鼠听神经损害研究

目的建立实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)大鼠模型,研究其脑干听觉诱发电位(brainstem auditory evoked potential,BAEP)Ⅰ波和听神经组织学改变。方法40只Wistar大鼠分为实验组和对照组,实验组(EAE组)用含卡介苗的豚鼠全脊髓匀浆为抗原免疫Wistar大鼠建立EAE动物模型,对照组用生理盐水混合完全福氏佐剂注射,采用诱发电位仪检测EAE组急性发病的大鼠和对照组大鼠的BAEP,通过透射电镜观察发病大鼠听神经外周段组织学改变。结果急性发病实验组大鼠BAEP的Ⅰ波潜伏期较对照组显著延长(P<0.01)。透射电镜下观察听神经外周段髓鞘松散,板层分离,局部变薄,呈不完全性脱髓鞘改变,轴突完整。对照组大鼠未见组织学改变。结论急性发病EAE大鼠听神经外周段脱髓鞘可能引起脑干听觉诱发电位Ⅰ波的改变。     

Anti-endothelial cell antibody and immune complexes in the sera of animals with acute experimental allergic encephalomyelitis and chronic relapsing experimental allergic encephalomyelitis

Blood-brain barrier (BBB) injury occurs in both acute and chronic relapsing experimental allergic encephalomyelitis (EAE). Sera from animals in which these forms of EAE had been induced were examined for anti-endothelial cell antibodies and immune complexes by enzyme-linked immunosorbent assay (ELISA) using either cultured endothelial cells or Raji cells. IgG binding to endothelial cells was significantly increased in the sera of animals with acute EAE and chronic relapsing EAE, compared to controls. Increased levels of circulating immune complexes were also detected in the sera of some animals with chronic relapsing EAE, especially those in an exacerbation. It is suggested that the anti-endothelial cell antibody and immune complexes detected may play pathogenetic roles in the destruction of the BBB in EAE.     

The FcRgamma chain is not essential for induction of experimental allergic encephalomyelitis (EAE) or anti-myelin antibody-mediated exacerbation of EAE

Macrophages are considered essential mediators in multiple sclerosis (MS) pathogenesis, presumably through myelin phagocytosis and release of inflammatory mediators. Macrophages and microglia express activating Fcgamma receptors (FcgammaRI and FcgammaRIII), which depend on the FcRgamma chain for surface expression and signaling. In MS lesions, crosslinking of FcgammaR by immunoglobulins (IgG) directed against myelin may enhance myelin phagocytosis and inflammation. We studied the role of FcgammaR and anti-myelin antibodies in MOG35-55-induced experimental allergic encephalomyelitis (EAE) in C57BL/6 mice, a model of MS-like disease. Incidence and severity of EAE were similar in FcRy chain-/- (FcRgamma-/-) and wild-type (wt) mice, albeit with delayed onset in FcRgamma-/- mice. This demonstrates that the FcRy chain is not essential for induction of EAE, but that FcRgamma signaling may contribute to the preclinical phase. The role of FcgammaR in antibody-mediated demyelination was addressed by injection of anti-myelin antibodies (Z12 mAb) at onset of MOG35-55-induced EAE. Injection of Z12 mAb rapidly reduced survival time in both wt and FcRgamma-/- mice, demonstrating that antibody-mediated exacerbation of EAE is independent of the FcRgamma chain. Interestingly, Z12-induced exacerbation of inflammation and demyelination persisted longer in wt than FcRgamma-/- mice, suggesting that IgG-FcgammaR interactions may contribute to a sustained pathologic effect of anti-myelin antibodies in the CNS.     

Morphological changes of the thymus and spleen with acute experimental allergic encephalomyelitis (EAE) in Lewis rats

Nervous and immune systems are thought to have an important role in host defence mechanism. Recent reports have suggested the presence of common antigens between nervous and immune systems. In this paper, we described the interactions between nervous and immune systems observed in an acute EAE model of Lewis rats. Seventeen Lewis rats were immunized by homogenized brain tissue in complete Freund adjuvant. Acute EAE rats were developed in 10 rats, all of which died within 25 days after the immunization. The thymus of these rats showed marked atrophy and histological examination revealed severe de generation of the lymphocytes and infiltration of macrophage showing phagocytosis in the thymus. The weight of spleen was also decreased, but no remarkable changes were observed microscopically except slight reduction of the white pulp. The fact that the immune reactions was induced not only in the nervous system but also in the thymus of EAE animals may suggest the possibility of common antigen between oligodendroglia and lymphocytes. This also suggest the close interaction between nervous system and immune system. However, the reactions of the thymus and spleen were different in this experiment, and further studies are necessary to evaluate the mechanism of these immune reactions.     

Converting relapsing remitting to secondary progressive experimental allergic encephalomyelitis (EAE) by ultraviolet B irradiation

We induced experimental allergic encephalomyelitis (EAE) in SJL/J mice, an animal model for multiple sclerosis (MS), using myelin oligodendrocyte glycoprotein (MOG)_92–106 peptide, following ultraviolet (UV) irradiation. While all control mice developed relapsing–remitting (RR)-EAE, UV irradiation induced secondary progressive (SP)-EAE in some of the mice. Although mild demyelination was observed with T cell infiltration in RR-EAE, large demyelinating lesions developed in SP-EAE with massive macrophage and neutrophil infiltration and immunoglobulin deposition, but with little T cell infiltration. UV irradiation induced higher anti-MOG antibody responses. In SP-EAE, lymphoproliferative responses and interferon-γ production were decreased without alteration of interleukin-4.     

Selective temporal and regional alterations of Nogo-A and small proline-rich repeat protein 1A (SPRR1A) but not Nogo-66 receptor (NgR) occur following traumatic brain injury in the rat

Axons show a poor regenerative capacity following traumatic central nervous system (CNS) injury, partly due to the expression of inhibitors of axonal outgrowth, of which Nogo-A is considered the most important. We evaluated the acute expression of Nogo-A, the Nogo-66 receptor (NgR) and the novel small proline-rich repeat protein 1A (SPRR1A, previously undetected in brain), following experimental lateral fluid percussion (FP) brain injury in rats. Immunofluorescence with antibodies against Nogo-A, NgR and SPRR1A was combined with antibodies against the neuronal markers NeuN and microtubule-associated protein (MAP)-2 and the oligodendrocyte marker RIP, while Western blot analysis was performed for Nogo-A and NgR. Brain injury produced a significant increase in Nogo-A expression in injured cortex, ipsilateral external capsule and reticular thalamus from days 1-7 post-injury (P < 0.05) compared to controls. Increased expression of Nogo-A was observed in both RIP- and NeuN positive (+) cells in the ipsilateral cortex, in NeuN (+) cells in the CA3 region of the hippocampus and reticular thalamus and in RIP (+) cells in white matter tracts. Alterations in NgR expression were not observed following traumatic brain injury (TBI). Brain injury increased the extent of SPRR1A expression in the ipsilateral cortex and the CA3 at all post-injury time-points in NeuN (+) cells. The marked increases in Nogo-A and SPRR1A in several important brain regions suggest that although inhibitors of axonal growth may be upregulated, the injured brain is also capable of expressing proteins promoting axonal outgrowth following TBI.