实验性变态反应性脑脊髓炎发病初期血脑屏障的形态学变化

<正> 多发性硬化(multiple sclerosis,MS)是以脑脊髓白质损伤为主的中枢神经系统炎性脱髓鞘疾病。实验性变态反应性脑脊髓炎(experimental allergic enceph-     

基质金属蛋白酶9在实验性自身免疫性脑脊髓炎大鼠发病过程中的动态表达

目的通过检测基质金属蛋白酶-9(MMP-9)在EAE大鼠发病过程中不同阶段外周血和中枢神经系统中的表达水平及动态变化,以探讨其在多发性硬化发病过程中的作用及机制。方法 Wistar雌性大鼠80只,分为模型组(EAE组)、完全福氏佐剂组(CFA组),分别于免疫后6天、8天、10天、12天、14天、16天、18天及20天取视交叉处脑组织和脊髓腰膨大段行HE染色观察炎性细胞的浸润状况;免疫组化法检测脑和脊髓组织中MMP-9的表达,酶联免疫吸附实验测定血清中MMP-9的含量。结果 EAE组大鼠脊髓和脑组织中MMP-9阳性细胞数及血清中MMP-9水平均显著高于同期CFA组(P<0.05);EAE 10天组与EAE 6天、14天、18天及20天组比较,大鼠脊髓腰膨大处MMP-9阳性细胞数较多(P<0.05);EAE 12天组与EAE 6天组、8天组、14天组、16天组、18天组及20天组比较,大鼠脑组织视交叉处MMP-9阳性细胞数较多(P<0.05);EAE 12天组与EAE 6天组、8天组、10天组、14天组、16天组、18天组及20天组相比较,大鼠血清中MMP-9含量较高(P<0.05)。结论在EAE大鼠发病前期即有中枢组织内MMP-9的高表达,且EAE大鼠脊髓中MMP-9的高表达要早于脑组织和外周血,但MMP-9表达的高峰在第12天与病理变化和疾病进展是同步的。     

T淋巴细胞增殖在实验性变态反应性脑脊髓炎发病中的作用

目的　探讨T淋巴细胞增殖在实验性变态反应性脑脊髓炎 (EAE)发病中的作用。方法　观察Lewis大鼠主动EAE临床症状和T淋巴细胞增殖。结果　EAE临床发病过程中 ,T淋巴细胞增殖旺盛。结论　EAE时 ,活化的T淋巴细胞可以通过BBB进入CNS ,浸入CNS的淋巴细胞能产生破坏性的细胞因子TNF α及某些酶类 ,推测T淋巴细胞增殖活跃 ,在导致EAE发病过程中起着至关重要的作用。     

基质金属蛋白酶-9在实验性自身免疫性脑脊髓炎大鼠的表达

目的 探讨基质金属蛋白酶-9（matrix metalloproteinase-9．MMP-9）在多发性硬化（MS）动物模型-实验性自身免疫性脑脊髓炎（experimental autoimmune encephalomyelitis．EAE）大鼠脑脊髓中的表达和作用。方法 应用豚鼠脑脊髓匀浆与完全福（氏）佐剂（CFA）混匀于Wistar大鼠颈背部皮下注射诱导建立EAE模型．同时将单独注射CFA或NS的大鼠设为对照组。采用半定量免疫组化方法检测Wistar大鼠发病后72h内及7、14d不同时间段脑脊髓中MMP-9的表达．并与对照组比较。结果 经免疫组化方法检测结果显示EAE大鼠脑脊髓内有特异的MMP-9表达．与对照组比较差异有显著性。发病72h内MMP-9表达高于发病后7d（P〈0．05）及发病后14d（P〈0．01）。结论 EAE大鼠脑脊髓的单核淋巴样细胞有特异MMP-9表达．且发病早期MMP-9阳性细胞率高于发病后期、恢复期及对照组．提示MMP-9与EAE发病有关。     

雌激素对实验性变态反应性脑脊髓炎大鼠发病影响的研究

目的探讨雌激素对实验性变态反应性脑脊髓炎(EAE)的影响。方法将30只大鼠随机分为EAE组及大、小剂量雌激素干预组,每组10只,制作EAE模型,大、小剂量雌激素干预组分别给予皮下注射苯甲酸雌二醇1 mg/kg/d、250μg/kg/d,连续10 d,观察各组的发病情况并采用HE染色观察脑和脊髓组织病理变化。结果大、小剂量雌激素干预组的临床症状均较EAE组轻,表现为发病率减少、潜伏期延长、进展期缩短、高峰期神经功能损害较轻。病理切片提示雌激素干预组脑和脊髓炎症细胞浸润明显减少,其中以大剂量组更明显。结论雌激素对多发性硬化动物模型EAE具有保护作用,且与剂量相关。     

Wistar大鼠实验性变态反应性脑脊髓炎的模型建立

目的:建立Wistar大鼠多病程实验性变态反应性脑脊髓炎(EAE)的动物模型,并进行病理学研究,为多发性硬化(MS)的研究提供实验依据。方法:以豚鼠全脊髓匀浆(GPSCH)为抗原免疫Wistar大鼠建立EAE的动物模型,并在光镜下观察不同发病类型EAE的病理改变。结果:根据病理和临床表现可将Wistar大鼠EAE模型分为5种发病形式:急性型、缓解-复发型、持续进展型、良性型和隐匿型。光镜下可见不同发病时期的EAE的病理改变有所不同,但都以血管"袖套"状改变、脑室周围及白质脱髓鞘改变为主,伴有神经元肿胀变性。结论:首次建立了Wistar大鼠多病程EAE,是研究多发性硬化的理想动物模型。     

载脂蛋白E拟肽对实验性变态反应性脑脊髓炎小鼠基质金属蛋白酶-9和基质金属蛋白酶组织抑制因子-1表达的影响

目的探讨载脂蛋白E(Apo E)拟肽对实验性变态反应性脑脊髓炎(EAE)小鼠基质金属蛋白酶-9(MMP-9)和基质金属蛋白酶组织抑制因子-1(TIMP-1)表达的影响。方法将30只雌性C57BL/6J小鼠随机分为Apo E拟肽组、EAE组和正常组,每组10只小鼠。EAE模型通过以髓鞘少突胶质细胞糖蛋白多肽35-55为抗原诱导。Apo E拟肽组在免疫后第2 d到30 d每隔2 d按5 mg/(kg·d)背部皮下注射Apo E拟肽。EAE组和正常组均以等体积生理盐水替代。免疫后第0~35 d每日对小鼠进行神经功能评分。免疫后第35d解剖小鼠,分离大脑和脊髓并行HE染色。采用免疫组化染色法检测各组小鼠大脑、脑干和脊髓的MMP-9和TIMP-1的表达。结果正常组小鼠均未发病。Apo E拟肽组、EAE组的小鼠全部发病,但各有1只小鼠发病后死亡。Apo E拟肽组与EAE组的发病潜伏期差异无统计学意义(P=0.72)。Apo E拟肽组的神经功能评分在峰值和慢性期(第35 d)均明显低于EAE组(均P0.05)。HE染色示,正常组未见炎症细胞浸润;EAE组小鼠大脑、脑干和脊髓均有不同程度的炎性细胞浸润,以脑干和脊髓较为明显;Apo E拟肽组小鼠CNS炎性细胞浸润相对于EAE组明显减少。EAE组小鼠大脑、脑干和脊髓的MMP-9表达均高于正常组(均P0.05)。Apo E拟肽组小鼠大脑和脊髓的MMP-9表达要明显低于EAE组(均P0.05),其中Apo E拟肽组小鼠中脑和脊髓的MMP-9表达与正常组相比无明显差异(均P0.05)。正常组小鼠脊髓TIMP-1的表达明显高于EAE组和Apo E拟肽组(均P0.05)。而Apo E拟肽组与EAE组小鼠大脑、脑干和脊髓TIMP-1表达的差异均无统计学意义(均P0.05)。结论 Apo E拟肽能通过抑制大脑和脊髓MMP-9的表达改善EAE小鼠的症状。     

多病程Wistar大鼠实验性变态反应性脑脊髓炎的病理研究

目的:在建立wistar大鼠多病程实验性变态反应性脑脊髓炎(EAE)的动物模型的基础上,进行病理学研究,探讨不同发病类型EAE的基本病理改变如炎细胞浸润、脱髓鞘和轴索损伤等方面的差别,为多发性硬化(MS)的研究提供实验依据。方法:以豚鼠全脊髓匀浆(GPSCH)为抗原免疫Wistar大鼠建立EAE的动物模型,进行常规HE染色、Weil髓鞘染色和改良的Bielschowsky,并行GFAP免疫组化染色,观察不同发病类型EAE的病理政变。结果:根据病理和临床表现可将Wistar大鼠EAE模型分为5种发病形式:急性型、缓解-复发型、持续进展型、良性型和隐匿型。光镜下可见不同发病时期的EAE的病理改变有所不同,急性型EAE炎症浸润明显,尤脱髓鞘改变,缓解-复发型和持续进展型EAE髓鞘脱失和轴索损伤更明显,且有陈旧病灶周围的星型胶质细胞增生,而新发病灶无此表现,良性型EAE则改变接近正常。结论:首次建立了Wistar大鼠多病程EAE,且病理证实不同类型EAE的炎细胞分布、髓鞘脱失及轴索损伤等基本病理改变是不同的,它具有人类MS的许多发病特点,其中多病程的发病形式和主要病理特点与MS极其相似,是理想的MS动物模型。     

调节性T细胞在实验性变态反应性脑脊髓炎和多发性硬化发病中的作用

调节性T细胞(regulatory T cells,Treg)是一类具有免疫调节功能的T细胞哑群.近年来,Treg细胞在实验性变态反应性脑脊髓炎(experimental allergical encephalomyelitis,EAE)、多发性硬化(multiple sclerosis,MS)发病中的作用越来越受到关注,小鼠Treg细胞缺失可导致特异性自身免疫性疾病,增加Treg细胞的功能可以减轻或抑制EAE.最近的研究结果表明,MS本身也伴随着成熟Treg细胞的受损或功能障碍.     

基质金属蛋白酶与实验性自身免疫性脑脊髓炎的免疫屏障

实验性自身免疫性脑脊髓炎（experimental autoimmune cncephalomyelitis，EAE）是一种主要由T细胞介导的，以中枢神经系统（CNSl内小血管周围单个核细胞浸润及白质髓鞘脱失为特征的自身免疫性疾病。EAE炎性脱髓鞘的过程有许多重要的炎性细胞因子参与。基质金属蛋白酶（matrix metalloproteinase，MMPs）尤其是MMP-9、MMP-2是引起血脑屏障（blood brain barrier，BBB）的免疫屏障破坏，炎性细胞浸润以及介导细胞外基质（ECM）和髓磷脂降解的重要因素。[第一段]     

MMP-9和MMP-2在多病程大鼠EAE发病过程中的变化

目的:研究MMP-2和MMP-9在多病程EAE不同类型间的变化,探讨MMPs在MS发病过程中的作用机制。方法:建立多病程大鼠EAE模型,以免疫组化的方法检测MMP-2、MMP-9在不同类型EAE中的表达及分布。结果:MMP-2、-9在EAE中的表达是一致的,不同类型EAE表达MMP-2、-9是不同的。1急性型EAE:炎细胞、血管内皮细胞及细胞外基质、脑膜细胞均呈阳性表达。2缓解-复发型EAE:活动性病灶呈阳性表达,非活动性病灶呈阴性表达。3持续进展型EAE:与缓解复发型EAE表达类似,但呈阳性表达的血管内皮细胞数目多于缓解-复发型。4良性型EAE:只有部分胶质细胞呈阳性表达。5隐匿型EAE-多数病灶呈轻度表达。结论:1 MMP-2、MMP-9在EAE中均有表达。2不同类型EAE中MMP-2、MMP-9的表达是不同的,与病理改变和疾病进展是一致和同步的。     

Astrogliosis in EAE spinal cord: derivation from radial glia, and relationships to oligodendroglia

A prominent feature of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE) is the accumulation of enlarged, multipolar glial fibrillary acidic protein (GFAP) and brain lipid binding protein (BLBP) immunoreactive astroglia within and at the margins of the inflammatory demyelinative lesions. Whether this astrogliosis is due to both astroglial hyperplasia and hypertrophy or solely to astroglial hypertrophy is controversial. We now report that coincident with the first appearance of inflammation and clinical deficits in mice with myelin oligodendrocyte glycoprotein peptide (MOG peptide)-induced EAE, the radially oriented, bipolar, GFAP, and BLBP positive cells (adult radial glia) present in normal spinal cord white matter undergo mitosis and phenotypic transformation to hypertrophic astroglia. To facilitate visualization of relationships between these hypertrophic astroglia and dying and regenerating oligodendroglia, we used mice that express enhanced green fluorescent protein (EGFP) in cells of the oligodendroglial lineage. During the first week after onset of illness, markedly swollen EGFP+ cells without processes were seen within lesions, whereas EGFP+ cells that expressed immunoreactive cleaved caspase-3 were uncommon. These observations support the hypothesis that necrosis contributes to oligodendroglial loss early in the course of EAE. Later in the illness, EGFP+ cells accumulated amongst hypertrophic astroglia at the margins of the lesions, while the lesions themselves remained depleted of oligodendroglia, suggesting that migration of oligodendroglial lineage cells into the lesions was retarded by the intense perilesional gliosis.     

The distribution of immunoglobulins and albumin in the central nervous system in acute experimental allergic encephalomyelitis

Experimental allergic encephalomyelitis (EAE) was induced in young male Lewis rats. Immunohistochemical visualisation of albumin and IgG in the nervous tissue was performed at intervals after induction. The results were correlated to the histological appearance of the tissue. Albumin appeared in the tissue about day 10, 1-2 days before IgG. Within one day both proteins spread from sharply defined perivascular subpial locations to diffuse distribution throughout the tissue. Cellular inflammation was not seen until 3-4 days after extravasation of proteins. The cells also spread from perivascular locations to a diffuse infiltration of the tissue.     

Serum pro-inflammatory and anti-inflammatory cytokines and the pathogenesis of experimental autoimmune encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) as an experimental model of multiple sclerosis (MS) is characterized by demyelination, infiltration of inflammatory cells into the nervous system and dysregulation of serum inflammatory cytokines. We investigated the correlation of serum cytokines and other inflammatory markers with the EAE pathogenesis. After EAE induction, the levels of different serum cytokine/inflammatory mediators were measured. Furthermore, motor functions, myelination, and lymphocyte infiltration in EAE mice were also assessed. Our results revealed that the serum concentrations of T-helper 1 (Th1) and Th17 cytokines, interleukin (IL)-6, IL-1β, IL-1α and prostaglandin E2 in EAE mice were significantly higher than controls. The ratios of pro- to anti-inflammatory cytokines were different between the EAE and the control group. A statistically significant positive correlation was found between the IL-6/IL-10 ratio and the EAE severity, demyelination rate, and lymphocyte infiltration in EAE mice. Results indicate that the profiles of serum pro- and anti-inflammatory cytokines might be useful as biomarkers for monitoring the pathological manifestation of EAE. Furthermore, evaluating the dynamic interplay of serum cytokine levels and the correlation with pathogenic mechanisms of EAE may provide diagnostic and therapeutic insights for MS and some other inflammatory disorders.     

The role of oxidative stress in the pathogenesis of multiple sclerosis: The need for effective antioxidant therapy

Abstract. Accumulating data indicate that oxidative stress (OS) plays a major role in the pathogenesis of multiple sclerosis (MS). Reactive oxygen species (ROS), leading to OS, generated in excess primarily by macrophages, have been implicated as mediators of demyelination and axonal damage in both MS and experimental autoimmune encephalomyelitis (EAE), its animal model. ROS cause damage to cardinal cellular components such as lipids, proteins and nucleic acids (e. g., RNA, DNA), resulting in cell death by necrosis or apoptosis. In addition, weakened cellular antioxidant defense systems in the central nervous system (CNS) in MS, and its vulnerability to ROS effects may increase damage. Thus, treatment with antioxidants might theoretically prevent propagation of tissue damage and improve both survival and neurological outcome. Indeed, several experimental studies have been performed to see whether dietary intake of several antioxidants prevents or reduces the progression of EAE. Although a few antioxidants showed some efficacy in these studies, little information is available on the effect of treatments with such compounds in patients with MS. Well-designed clinical studies using antioxidant intake, as well as investigations based on larger cohorts studied over a longer periods of time, are needed in order to assess whether antioxidant intake together with other conventional treatments, might be beneficial in treating MS.     

树突状细胞在自身免疫性脑脊髓炎发病及耐受中作用的研究

目的　研究树突状细胞 ( dendritic cells,DCs)在实验性自身免疫性脑脊髓炎 ( experimental allergicencephalomyelitis,EAE)发病及免疫耐受中的作用。方法　诱导 EAE和口服耐受 EAE模型 ,采用流式细胞仪和直接免疫荧光法观察 EAE不同阶段、不同疾病程度外周淋巴结和中枢神经系统 ( CNS)中 DCs数量的变化。结果　免疫后第 4、7、9天外周淋巴结 DCs数量逐渐增多 ,第 9天脑、脊髓中开始有 DCs浸润 ,第 1 5天 DCs数量达高峰 ,第 1 5、2 3天 DCs数量与疾病症状严重程度相一致。口服耐受组与 EAE组差异无显著性。结论　外周淋巴结和 CNS局部 DCs在 EAE的发生、发展、转归尤其在触发发病中起重要作用。 DCs作为免疫反应的始动者 ,其数量变化在口服免疫耐受的产生机制中具次要作用     

Concordance and localization of maximal vascular permeability change and fibrin deposition in the central neuraxis of Lewis rats with cell-transferred experimental allergic encephalomyelitis

This study utilized Lewis rats and the cell-transfer form of experimental allergic encephalomyelitis (EAE) to focus on two central nervous system microvascular alterations known to be intimately associated with early clinical neurological signs of the disease, namely increased vascular permeability and deposition of fibrin. The main objective of the work was to define the degree of concordance and the anatomical localization of maximal vascular permeability change and fibrin deposition within the neuraxis of recipient rats during the earliest expression of clinical manifestations of the disease. Recipients were injected with predetermined doses of activated, myelin basic protein (MBP)-reactive syngeneic donor lymphoid cells and killed 6 or 7 days later when they exhibited the first clinical signs of disease. This experimental design allowed assessment of late occurring immune activities of a single population of MBP-reactive effector cells. Increased vascular permeability, expressed as mean extravascular blood equivalents (EVBE), attained maximum change, i.e., 16.04 +/- 5.07, within the lumbosacrococcygeal segment of the spinal cord (LSC). Maximal density of fibrin depositions, i.e., 192 +/- 94 deposits, also occurred within the LSC cord. The concordance of these two histoimmunopathological events, their tight linkage to early clinical signs of disease and the fact that both events attained maximal values within the LSC cord suggest that increased permeability and activation of the coagulation cascade may be prerequisite events for the expression of neurological signs of EAE.     

B7-2(CD86)协调刺激因子在急性实验性变态反应性脑脊髓炎(AEAE)发挥主导作用

目的　探讨协调刺激分子B7 1(CD80 )和B7 2 (CD86)在急性EAE发病过程中的作用。方法　观察抗B7 1和B7 2抗体在体外对淋巴细胞抗原特异性增殖反应和细胞因子分泌的抑制作用和在体内对EAE发生过程的影响。结果　抗B7 2抗体抑制PLP136 150抗原引起的特异性细胞增殖和IL 2产生 ,抗B7 2抗体处理过的淋巴母细胞诱导轻症被动EAE ;在主动EAE诱导的早期抗B7 2抗体虽能延缓发病时间但加重病情 ,可能与IL 4分泌不足有关 ;当临床EAE首发症状出现时 ,抗B7 2抗体减轻其临床表现。结论　协调刺激分子B7 2在AEAE发生过程中发挥重要作用