一种新的基质金属蛋白酶抑制剂ONO-4817对实验性自身免疫性脑脊髓炎的治疗作用

目的　探讨一种新合成的含氧肟酸的基质金属蛋白酶 ( MMP)抑制剂 ONO-481 7对实验性自身免疫性脑脊髓炎 ( EAE)的治疗效果。方法　给 EAE大鼠口服 ONO-481 7,观察临床症状、T淋巴细胞增殖以及血清肿瘤坏死因子 ( TNF) -α水平。结果　 ONO-481 7能显著改善 EAE临床症状 ( P <0 .0 1 ) ,同时明显抑制 T淋巴细胞增殖 ( P <0 .0 1 ) ,显著降低大鼠血清 TNF-α水平 ( P <0 .0 5 )。结论　研究表明 ,ONO-481 7通过抑制 MMPs活性、T淋巴细胞增殖和减少 TNF-α生成 ,进而能显著减轻血脑屏障 ( BBB)的破坏 ,又可抑制炎细胞浸润和髓鞘破坏 ,从而有效缓解 EAE。     

实验性变态反应性脑脊髓炎T淋巴细胞增殖和肿瘤坏死因子—α变化的意义

目的：探讨实验性变态反应性脑脊髓炎（EAE）T淋巴细胞增殖和肿瘤坏死因子（TNF）－α变化的意义。方法：观察Lewis大鼠主动免疫性EAE临床症状,T淋巴细胞增殖以及血清TNF－α水平,结果：EAE临床发病过程中,T淋巴细胞增殖活跃,血清TNF－α水平异常升高,并与临床症状呈正相关。结论：研究表明：EAE时,TI林巴细胞增殖旺盛和TNF－α生成增多。进而导致血脑屏障的破坏,炎细胞浸润,从而诱发EAE。     

TNF-α在实验性变态反应性脑脊髓炎中变化的意义

实验性变态反应性脑脊髓炎(EAE)是一种T淋巴细胞介导的中枢神经系统(CNS)免疫性疾病,与人类CNS疾患多发性硬化(MS)具有很多共同的特征.在MS和EAE发病机制中,血清肿瘤坏死因子-α(TNF-α)一直是人们所关注的因素.本文就EAE血清TNF-α的变化与临床症状关系进行研究,现报告如下.     

甲基强的松龙治疗实验性变态反应性脑脊髓炎的作用机制

目的:研究细胞因子、T细胞凋亡和淋巴细胞增殖在实验性变态反应性脑脊髓炎(EAE)形成中的作用及甲基强的松龙(MP)治疗EAE的作用机制。方法:采用人脑纯化的髓鞘碱性蛋白(MBP)与完全福氏佐剂免疫Lewis大鼠,建立EAE动物模型。用双抗体夹心ELISA法检测各组大鼠血清中IL-10、TNF-α、IFN-γ的含量:流式细胞仪检测外周血T细胞凋亡;3H-TdR释放法检测外周血淋巴细胞转化率。结果:与对照组比较,EAE组的外周血IFN-γ、TNF-α水平明显增高,IL-10水平明显降低,MP治疗后IFN-γ和TNF-α水平下降,IL-10浓度上调。MP还诱导外周血T细胞凋亡和抑制MBP致敏淋巴细胞增殖并呈剂量依赖性。结论:应用人MBP成功建立EAE大鼠模型,MP可能通过调节Th细胞因子格局、促进Th2细胞因子分泌、抑制MBP致敏淋巴细胞增殖及外周血T细胞凋亡而发挥治疗多发性硬化的作用。     

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。     

雌激素对实验性自身免疫性脑脊髓炎小鼠的抗炎机制

目的 探讨雌激素减轻实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)炎症反应的可能机制.方法 用MOG35-55多肽诱发60只EAE小鼠模型,做去卵巢术.分为治疗组(n=30)和对照组(n=30),治疗组予雌激素治疗.比较两组EAE小鼠的临床症状评分.取脑和脊髓,行H-E染色观察病理学改变,实时荧光定量PCR及ELISA检测EAE小鼠CNS中白细胞介素(interleukin,IL)-17、IL-23、肿瘤坏死因子α(tumor necrosis factor α,TNF-α)、干扰素γ(interferon γ,IFN-γ)、IL-4水平.结果 治疗组EAE小鼠与对照组相比临床症状减轻(P＜0.05);H-E染色显示治疗组炎细胞浸润减少(P＜0.05),实时荧光定量PCR及ELISA结果示治疗组CNS中IL-17、IL-23、TNF-α、IFN-γ表达降低而IL-4增加(P＜0.05).结论 雌激素可能通过降低IL-17、IL-23、TNF-α、IFN-γ,增加IL-4,从而减轻EAE小鼠炎症反应.     

胸腺五肽对实验性变态反应性脑脊髓炎的治疗机制探讨

目的:探讨胸腺五肽对实验性变态反应性脑脊髓炎的治疗效果及脑组织内肿瘤坏死因子-α水平的影响,为临床应用胸腺五肽提供理论依据。方法:成年雌性Wistar大鼠48只,随机分为正常对照组,EAE组,胸腺五肽治疗组。动态、对比观察实验各组在EAE诱导后7d、14d、21d时发病时间、发病率、病情程度、脑组织的病理变化及脑组织肿瘤坏死因子-α水平的改变。结果:胸腺五肽治疗组大鼠总发病率、发病程度与EAE组比较均明显降低(P<0.05),发病时间较EAE组比较平均延迟2.14d,但统计学差异不显著(P>0.05);脑组织病理改变较EAE组病灶出现晚且损伤减轻;对照组大鼠脑组织TNF-α蛋白有一定量的表达。EAE组大鼠TNF-α表达量显著高于对照组(P<0.01)和治疗组。胸腺五肽治疗组较对照组有所升高(无统计学差异,P>0.05),但比EAE组大鼠脑组织TNF-α含量明显降低,P<0.05)。结论:胸腺五肽能显著降低EAE大鼠的发病率、病情程度和减轻脑组织损伤,对EAE发病起到了治疗作用;通过脑组织TNF-α的含量变化表明,其发挥疗效的机制在于胸腺五肽显著降低了TNF-α蛋白的表达水平,减低了脑内炎症反应的程度所致。     

血清肌钙蛋白T及生化标志物联合检测对急性脑缺血性卒中的诊断意义

目的探讨肌钙蛋白T(cTnT)、甲状腺T3和肿瘤坏死因子(TNF-α))在急性脑缺血性卒中的作用及临床价值。方法 60例急性脑缺血性卒中患者和50例健康体检者作为研究对象,采用电化学发光免疫诊断仪测定血清cTnT和T3水平,采用酶联免疫吸附(ELISA)测定TNF-α水平。结果观察组患者cTnT、T3、TNF-α水平显著上升,与对照组比较有显著性差异(P0.05)。结论 cTnT、T3、TNF-α与急性脑缺血性卒中心肌损害和预后有关,检测这些指标对急性脑缺血性卒中诊断和预后具有重要价值。     

肿瘤坏死因子-α与抑郁症患者躯体疼痛症状相关性的研究

目的:探讨抑郁症患者血清肿瘤坏死因子-α(TNF-α)水平与伴有的躯体疼痛症状的关系。方法:对42例伴有躯体疼痛症状的抑郁症患者(研究组)和79例不伴有躯体疼痛症状的抑郁症患者(对照组)测定血清TNF-α水平,并采用汉密尔顿抑郁量表(HAMD-24)、汉密尔顿焦虑量表(HAMA)、视觉模拟评分(VAS)分别评估患者的抑郁症状、焦虑症状、躯体疼痛症状的程度,并进行相关性分析。结果:研究组血清TNF-α水平[(29.6±4.8)ng/L]显著高于对照组[(27.5±4.2)ng/L](t=2.491,P=0.027);经协方差分析调整影响因素后,两组血清TNF-α水平差异仍有统计学意义(F=3.855,P=0.036)。相关分析显示,研究组血清TNF-α水平与VAS评分、HAMD评分、HAMA评分呈正相关(r=0.362,P=0.018;r=0.408,P=0.006;r=0.336,P=0.038),对照组血清TNF-α水平与HAMD评分正相关(r=0.307,P=0.029)。结论:伴有躯体疼痛的抑郁症患者血清TNF-α水平增高,并可能与躯体疼痛症状的发生有关。     

急性EAE豚鼠脑白质TNF-α mRNA的表达及其实验性治疗

目的 探讨实验性变应性脑脊髓炎(EAE)豚鼠脑白质肿瘤坏死因子(TNF-α)mRNA的表达以及己酮可可碱、培高利特对其表达的影响。方法 采用免疫诱导方法制备EAE豚鼠模型,应用RT-PCR方法检测:EAE豚鼠脑白质TNF-α mRNA的表达。结果 EAE组豚鼠脑白质TNF-α mRNA的表达明显高于对照组(P<0.01),治疗组较EAE组表达下降(P<0.05)。结论 急性EAE豚鼠脑白质TNF-α mRNA的表达上调,与疾病的触发有关,己酮可可碱、培高利特对TNF-α mRNA的表达具有不同程度的抑制作用。     

甲基强的松龙及联合丹奥治疗缺血性脑血管病的临床研究

目的 :探讨免疫抑制剂甲基强的松龙 (MP)对急性脑梗塞的影响 ,及 MP与血栓素合成酶抑制剂丹奥对急性脑梗塞的交互作用。方法 :采用析因试验设计 ,观察 MP组、丹奥组、 MP联合组、对照组的肿瘤坏死因子α(TNF-α)、淋巴细胞转化率、胞浆酶、神经功能缺损评分及临床疗效的变化。结果 :MP组及 MP联合组的 TNF-α水平及淋巴细胞转化率明显下降 ,与丹奥组及对照组相比有显著性差异 ;MP组、丹奥组及 MP联合组治疗后的神经功能缺损评分下降 ,显效率增高 ,MP与丹奥有交互作用。结论 :免疫抑制剂 MP可减轻缺血性脑损伤 ,联合应用丹奥可显著改善缺血性脑血管病的预后 ,降低致残率     

Human tumor necrosis factor-alpha augments experimental allergic encephalomyelitis in rats

The effect of human recombinant tumor necrosis factor-alpha (TNF) on experimental allergic encephalomyelitis (EAE) was studied in Lewis rats. TNF was injected intraperitoneally at a daily dose of 1 x 10(3) or 2 x 10(4) U for 8 consecutive days from one day after sensitization with guinea-pig spinal cord in complete Freund's adjuvant. All rats in the control group developed clinical signs of EAE but recovered within 8 days after the onset. Injections of 2 x 10(4) U/day of TNF resulted in a significant prolongation of clinical EAE: clinical signs were sustained for up to 15 days after onset. Histologically, rats receiving 2 x 10(4) U/day of TNF had more severe cellular infiltrations in the spinal cord than controls. The augmentation of EAE was not found in rats receiving 1 x 10(3) U/day of TNF or TNF that had been neutralized with anti-TNF monoclonal antibody.     

Lactoferrin ameliorates symptoms of experimental encephalomyelitis in Lewis rats

Lactoferrin (LF) is a multifunctional protein present in secretory fluids of mammals and circulating neutrophils. Beside anti-inflammatory properties, LF was found to inhibit some autoimmune disorders. In this investigation we studied effects of oral administration of LF on experimental autoimmune encephalomyelitis (EAE) in Lewis rats. LF was given in drinking water as 0.25% solution beginning the day of elicitation of EAE or with a seven-day delay. The effects of LF were evaluated by the following criteria: clinical score, lymph node cell number, serum cytokine levels and histopathological changes. We found that LF treatment led to a significant acceleration of the recovery process, particularly on days 16–18 following elicitation of EAE. The delayed administration of LF was less effective in reducing the score of EAE. In addition, cell number of the inguinal lymph nodes of untreated EAE rats, almost 3 times higher as compared with control, naïve rats, was normalized by LF treatment. Furthermore, LF decreased elevated serum concentrations of tumor necrosis factor alpha and transforming growth factor beta. The histological analysis of the spinal cord revealed reduction in the number and size of inflammatory foci in LF-treated rats. In summary, treatment of EAE Lewis rats with LF reduced the clinical symptoms and accelerated the recovery of animals.     

Cytokine production in the central nervous system of Lewis rats with experimental autoimmune encephalomyelitis: dynamics of mRNA expression for interleukin-10, interleukin-12, cytolysin,tumor necrosis factor α and tumor necrosis factor β

The kinetics of mRNA expression in the central nervous system (CNS) for a series of putatively disease-promoting and disease-limiting cytokines during the course of experimental autoimmune encephalomyelitis (EAE) in Lewis rats were studied. Cytokine mRNA-expressing cells were detected in cryosections of spinal cords using in situ hybridization technique with synthetic oligonucleotide probes. Three stages of cytokine mRNA expression could be distinguished: (i) interleukin (IL)-12, tumor necrosis factor (TNF)-β (=lymphotoxin-α) and cytolysin appeared early and before onset of clinical signs of EAE; (ii) TNF-α peaked at height of clinical signs of EAE; (iii) IL-10 appeared increasingly at and after clinical recovery. The early expression of IL-12 prior to the expression of interferon-γ (IFN-γ) mRNA shown previously is consistent with a role of IL-12 in promoting proliferation and activation of T helper 1 (Th1) type cells producing IFN-γ. The TNF-β mRNA expression prior to onset of clinical signs favours a role for this cytokine in disease initiation. A pathogenic effector role of TNF-α was suggested from these observations that TNF-α mRNA expression roughly paralleled the clinical signs of EAE. This may be the case also for cytolysin. IL-10-expressing cells gradually increased to high levels in the recovery phase of EAE, consistent with a function in down-regulating the CNS inflammation. From these data we conclude that there is an ordered appearance of putative disease-promoting and -limiting cytokines in the CNS during acute monophasic EAE.     

Defective CD4T cell priming and resistance to experimental autoimmune encephalomyelitis in TNF-deficient mice due to innate immune hypo-responsiveness

We report here that tumor necrosis factor (TNF) deficiency causes innate hypo-responsiveness to a broad range of bacterial or viral constituents. In vivo hypo-responsiveness of TNF-deficient mice to mycobacteria results in defective CD4+ T cell priming to antigens administered in complete Freund's adjuvant (CFA). This deficiency is restored by supplementary mycobacteria. Furthermore, we show that even when self-reactive CD4+ T cell priming is fully restored, susceptibility of TNF-deficient mice to experimental autoimmune encephalomyelitis (EAE) depends on the co-administered pertussis toxin (PTx). TNF-deficient mice are completely resistant to EAE at sub-optimal doses of PTx, while supplementary PTx restores susceptibility. Therefore, TNF shows distinct functions in linking innate responsiveness to CD4+ T cell priming and to the induction of autoimmune disease.     

Type IV phosphodiesterase inhibition in experimental allergic encephalomyelitis of Lewis rats: sequential gene expression analysis of cytokines, adhesion molecules and the inducible nitric oxide synthase.

Type IV phosphodiesterase inhibitors are able to suppress EAE. To investigate the effects of this therapy in the central nervous system, we serially analyzed from days 7 to 17 postinoculation the gene expression pattern of tumor necrosis factor (TNF), lymphotoxin, interferon-gamma, interleukin-1beta, the inducible nitric oxide synthase (iNOs), interleukin-10, the vascular cell adhesion molecule-1 (VCAM-1) and the intercellular adhesion molecule-1 (ICAM-1) in the spinal cord of Lewis rats with actively induced EAE, treated with Rolipram. Treated rats had a delayed and milder disease, and reduced numbers of infiltrates in the nervous tissue. The gene expression profile was similar to that of untreated rats, although delayed, with no evidence of IL-10 upregulation during the observation period. The delayed inflammation was not associated with changes in the expression of VCAM-1 and ICAM-1. In peripheral blood mononuclear cells, TNF mRNA levels were decreased and interleukin-10 was unchanged. This therapy did not alter the proliferative ability of T lymphocytes against myelin basic protein. The encephalitogenic potential of splenocytes from treated animals was also unaffected. The high levels of both iNOs mRNA and nitric oxide (NO) found before the appearance of clinical signs, suggests that NO generation might be a contributing factor to the therapeutic benefit achieved by Rolipram in the rat.     

Prevention of chronic relapsing experimental autoimmune encephalomyelitis by soluble tumor necrosis factor receptor I

We have evaluated the effect of the type I (p-55, type β) soluble tumor necrosis factor receptor (sTNFrI) in an animal model of multiple sclerosis. Experimental autoimmune encephalomyelitis (EAE) was induced in SJL/J mice by adoptive transfer of T lymphocytes sensitized to myelin basic protein (MBP). sTNFrI completely blocked both clinical signs of disease and pathological changes that included CNS demyelination and inflammatory cell infiltration. Effective inhibition of disease expression was obtained using several different regimens of subcutaneous (s.c.) injection. These included daily doses starting at day 0, every other day injections starting at day 0, daily doses starting on day 4, and two doses separated by 12 h on day 1 and 2. Furthermore, treatment with sTNFrI for 15 days completely protected these animals from the recurrent episodes of disease normally associated with adoptively transferred EAE. These findings suggest that TNF plays a major causative role in EAE and that the sTNFrI may prove to be a useful therapeutic approach in multiple sclerosis.     

Suppressive activity of long-term myelin basic protein-specific SJL T cell lines

Myelin basic protein (MBP)-specific T cell lines derived from SJL mice lose the ability to transfer adoptively experimental allergic encephalomyelitis (EAE) after 5-6 restimulations with antigen in vitro. In order to test whether such lines were suppressive, non-encephalitogenic T cell lines were co-cultured with a freshly derived encephalitogenic T cell line. Following co-culture in the presence of MBP and irradiated syngeneic spleen cells the mixture was transferred adoptively to syngeneic recipients. Severe EAE was observed in recipients of the encephalitogenic cell line alone but not in animals which received the co-culture. A co-culture period was required as mixing the encephalitogenic and non-encephalitogenic T cell lines just prior to transfer was without effect. Not all non-encephalitogenic cell lines were found to be suppressive. Culture fluids from the suppressive, but not the non-suppressive lines were found to inhibit MBP-driven proliferation of T cell clones and encephalitogenic lines in vitro. Nineteen of 55 MBP-specific T cell clones derived from suppressive lines were found to elaborate the suppressive supernatant activity. The suppressive effect was not antigen-specific since the same culture supernatants inhibited proliferation of an ovalbumin-specific SJL T cell clone. The suppressive effect became apparent only after T cell lines had lost encephalitogenicity and was not mediated by tumor necrosis factor, lymphotoxin or prostaglandin.