地塞米松对实验性变态反应性脑脊髓炎小鼠胸腺细胞凋亡的影响

目的 探讨地塞米松对实验性变态反应性脑脊髓炎（ＥＡＥ）小鼠胸腺细胞凋亡的影响。方法 采用昆明小鼠建立ＥＡＥ模型,于处死前２４小时腹腔注射米松,通过荧光染色法、原位末端标记法和电镜法观察地塞米松处理组,自然病程组和对照组的小鼠胸腺细胞凋亡。结果 地塞米松处理组的胸腺细胞凋亡率明显高于自然病程组和正常对照组（Ｐ〈０．０１）;且电镜发现有典型的凋亡改变。结论 地塞米松可明显增强ＥＡＥ小鼠胸腺细胞凋亡。     

β淀粉样蛋白诱导脑内神经元凋亡的研究

目的 探讨凋亡机制在β－淀粉样蛋白（β－ａｍｙｌｏｉｄ ｐｒｏｔｅｉｎ,Ａβ）脑内致病作用中的意义。方法 用微量注射器将Ａβ１－４０注射到大鼠右侧海马Ｃ工区诱发Ａβ在脑内该区域的沉积。７天后,用ＨＥ染色、ＴＵＮＥＬ法及透射电镜检测该区细胞凋亡,用免疫组化ＳＡＢＣ法检测Ｂａｘ／Ｂｃｌ－２的表达。结果 在Ａβ组右侧少我ＨＥ１区ＨＥ、ＴＵＮＥＬ染色及电镜均发现大量凋亡细胞,而假手术对照组和生理盐水对照组     

正常老年人与阿尔茨海默病脑内生长抑素 神经元分布及其与细胞凋亡的关系

目的探讨正常老年人与阿尔茨海默病（Ａｌｚｈｅｉｍｅｒｓｄｉｓｅａｓｅ,ＡＤ）患者不同区域大脑皮层中生长抑素（Ｓｏｍａｔｏｓｔａｔｉｎ,ＳＯＭ）阳性神经元数量是否存在差异,以及这些神经元是否以细胞凋亡的方式死亡或消失。方法采用免疫组织化学方法观察ＳＯＭ阳性神经元;采用ＴＵＮＥＬ法观察细胞凋亡。结果正常老年人及ＡＤ病人大脑皮层不同区域内的ＳＯＭ阳性神经元的数量未见显著性差异,但可见细胞凋亡。而ＳＯＭ阳性神经元与ＴＵＮＥＬ阳性细胞并不完全重叠出现。结论正常老年人及ＡＤ病人大脑皮层内ＳＯＭ阳性神经元均较少,但两者之间无明显差异。细胞凋亡与ＳＯＭ阳性神经元之间似无明显相关性。     

重症肌无力胸腺和外周血抗原特异性单个核细胞IFN—？…

目的 为了解重症肌无力（ＭＧ）胸腺细胞免疫状态,对１０例行胸腺摘降（Ｔｘ）的ＭＧ患者及４例先天性心脏病并行开胸手术患者的胸腺和外周血单个核细胞经特异性抗原乙酰胆碱受体（ＡＣｈＲ）刺激后,其细胞ＩＦＮ－γ和ＩＬ－４的ｍＲＮＡ转录和细胞培养上清液中的蛋白表达情况进行了检测。方法 采用ＲＴ－ＰＣＲ结合狭缝印迹杂交检测ＩＦＮ－γ,ＩＬ－４ｍＲＮＡ转录,采用ＥＬＩＳＡ检测ＩＦＮ－γ和ＩＬ－４的表达情况。结果     

EAN血清可溶性细胞间粘附分子—1的测定

目的探讨血清可溶性细胞间粘附分子－１（ｓＩＣＡＭ－１）在实验性变态反应性神经炎（ＥＡＮ）中的作用。方法用兔坐骨神经匀浆加完全福氏佐剂（ＣＦＡ）免疫大鼠，建立ＥＡＮ模型；同时用抗细胞间粘附分子－１（ＩＣＡＭ－１）单克隆抗体注入大鼠体内后再诱导ＥＡＮ；观察自然病程组、抗体注射组及对照组的发病情况与病理特点；应用双抗体夹心ＥＬＩＳＡ法检测不同发病程度ＥＡＮ动物血清中ｓＩＣＡＭ－１的浓度。结果抗体注射组发病率及发病程度明显低于自然病程组；自然病程组ｓＩＣＡＭ－１的浓度高于抗体注射组及对照组；ＥＡＮ发病程度与ｓＩＣＡＭ－１的浓度呈正比。结论ｓＩＣＡＭ－１与ＥＡＮ的发病关系密切，ｓＩＣＡＭ－１的测定是观测自身免疫性疾病的一个有用的指标；抗ＩＣＡＭ－１抗体能够减轻或预防ＥＡＮ的发生。     

AChRAb阳性和阴性重症肌无力患者IL—4和IFN—γ…

将行胸腺切除术的重症肌无力（ＭＧ）患者分成乙酰胆碱受体抗体（ＡＣｈＲＡｂ）阳性和阴性２组，采用免疫酶点法检测其外周血、骨髓和胸腺白细胞介素４－（ＩＬ－４）分泌细胞和干扰素－γ（ＩＦＮ－γ）分泌细胞的数量，结果表明ＡＣｈＲＡｂ阳性组其外周血和骨髓中ＩＬ－４和ＩＦＮ－γ分泌细胞数量均显著高于ＡＣｈＲＡｂ阴性组（Ｐ〈０．０５），而胸腺细胞两组间差异无显著性意义（Ｐ〉０．０５）。提示ＩＬ－４和ＩＦＮ－γ在     

β淀粉样蛋白诱导脑内神经元调亡及褪黑素的保护作用

目的 探讨凋亡机制在β－淀粉样蛋白（Ａβ）服内致阿尔茨海默病（ＡＤ）作用中的意义及褪黑素（ＭＴ）对Ａβ脑内神经毒性的干预效果。地将Ａβ１－４０微量注射至大鼠右侧海马ＣＡ１区,７天后,用尼氏染色检测神经元丢失,ＨＥ染色、ＴＵＮＥＬ染色及透射电镜检测细胞凋亡,用免疫组织化ＳＡＢＣ法检测Ｂａｘ／Ｂｃｌ－２表达。结果 Ａβ组右侧海马ＣＡ１区发现大量凋亡细胞,而假手术对照组和生理盐水对照组未发现细胞凋亡;Ａ     

ABC－ELISA法检测重症肌无力患者三种自身抗体的研究

本文利用ＡＢＣ－ＥＬＩＳＡ法检测了９７例ＭＧ病人血清内三种抗体：ＡｃｈＲａｂ、Ｐｒ－Ｍａｂ、ＣＡＥａｂ。结果发现：（１）全身型ＡｃｈＲａｂ、ｐｒ－Ｍａｂ阳性率明显高于眼肌型（Ｐ＜０．０１）；ＡｃｈＲａｂ与Ｐｒ－ＭａｂＰ／Ｎ值呈线性正相关（ｒ＝０．７９７Ｐ＜０．０１）。（２）合并胸腺异常者ＣＡＥａｂ阳性率为８４．２％，明显高于对照组（Ｐ＜０．０１）；此组病人ＡｃｈＲａｂ与ＣＡＥａｂＰ／Ｎ值呈显著正相关，Ｐｒ－Ｍａｂ与ＣＡＥａｂＰ／Ｎ值呈非常显著正相关。（ｒ＝０．５１２和ｒ＝０．５９８Ｐ＜０．０１）。（３）８例病人作了治疗前后抗体检测。激素治疗后或切除异常胸腺后，抗体滴度多数下降或有转阴趋势。     

帕金森病小鼠黑质细胞凋亡的实验研究

目的探讨细胞凋亡在帕金森病发病机制中的作用。方法给Ｃ５７ＢＬ小鼠腹腔注射不同剂量、不同时限的１甲基４苯基１，２，３，６四氢吡啶（ＭＰＴＰ），在用ＭＰＴＰ之前，给Ｃ５７ＢＬ小鼠口服咪多吡（Ｅｌｄｅｐｒｙｌ）或Ｒｉｌｕｚｏｌｅ，分别用ＤＮＡ末端标记法（ＴＵＮＥＬ）和流式细胞术（ＦＡＣＳ）对小鼠黑质细胞检测凋亡，并用同样方法检测分别用等浓度的１甲基４苯基吡啶离子（ＭＰＰ＋）及ＭＰＴＰ处理的ＰＣ１２细胞。结果３０ｍｇ／ｋｇ体重ＭＰＴＰ连续应用３天以上诱发黑质细胞凋亡，而１０ｍｇ／ｋｇ体重ＭＰＴＰ无致凋亡作用。咪多吡预处理可防止ＭＰＴＰ诱发的黑质细胞凋亡；Ｒｉｌｕｚｏｌｅ能减轻ＭＰＴＰ诱发的黑质细胞凋亡程度。２０μｍｏｌ／Ｌ浓度的ＭＰＰ＋使ＰＣ１２细胞发生凋亡，等浓度的ＭＰＴＰ对ＰＣ１２细胞无致凋亡作用。结论帕金森病发病机制可能与细胞凋亡有关。ＭＰＴＰ在体内转化为ＭＰＰ＋导致黑质细胞凋亡。     

重症肌无力患者血清连接素抗体的检测及其临床意义

目的 探讨连接素（ｔｉｔｉｎ或ｃｏｎｎｅｃｔｉｎ）抗体９＝（ｔｉｔｉｎａｂ）与重症肌无力（ＭＧ）的关系。方法 应用基因工程合成重组重症肌无力胸腺瘤特异性３００００抗原（ＭＧＴ－３０蛋白）,并采用酶联免疫吸附法（ＥＬＩＳＡ）检测１４１例不同胸腺病理类型的ＭＧ患者（ＭＧ组）、２６５名健康对照者（ＮＣ组）和３６例非ＭＧ其他疾病患者（ＮＭＧ组）血清中ｔｉｔｉｎａｂ水平,同时检测ＭＧ患者血清中乙酰胆碱受体抗体（ＡＣｈＲａｂ）、突触前膜受体抗体（ＰｒｓｍＲａｂ）水平。结果 ＭＧ组血清中ｔｉｔｉｎａｂ水平明显高于ＮＣ组和ＮＭＧ组（均Ｐ〈０．０１）;阳性率以ＭＧ伴胸腺瘤（ＭＧＴ）组最高（８３．７％）,ＭＧ伴胸腺萎缩（ＭＧＡ）组其次（４３．２％０,而ＭＧ伴胸腺增生（ＭＧＨ）组及ＭＧ胸腺正常（ＭＧＮ）组患者血清中ｔｉｔｉｎａｂ均为     

硫脂致敏豚鼠血清诱导体外培养的施万细胞凋亡及其机制

目的进一步探讨硫脂在周围神经髓鞘损害过程中的作用机制。方法在植块式体外培养的施万细胞（Ｓｃｈｗａｎｎｃｅｌｓ，ＳＣｓ）培养液中，加１０％硫脂致敏的豚鼠血清１２小时后，利用末端标记程序性细胞死亡检测法和扫描、透射电镜观察凋亡的ＳＣｓ数及ＳＣｓ的形态变化，并通过免疫细胞化学和原位杂交的方法观察ＳＣｓ中凋亡基因Ｆａｓ蛋白及其ｍＲＮＡ的表达。结果加硫脂致敏豚鼠血清后，ＳＣｓ胞浆中Ｆａｓ蛋白及其ｍＲＮＡ的表达较对照组明显增强，凋亡的ＳＣｓ数显著增加，扫描、透射电镜检查显示，部分ＳＣｓ出现凋亡的早期病理改变。结论硫脂致敏豚鼠血清具有诱导体外培养的ＳＣｓ凋亡的作用。     

The therapeutic effects of ginkgolides in Guillain-Barré syndrome and experimental autoimmune neuritis

BackgroundGuillain-Barré syndrome (GBS) is an acquired immune-mediated inflammatory peripheral neuropathy. The immune regulation of ginkgolides have been revealed in recent years. We herein investigate the potential therapeutic effects of ginkgolides both on GBS and its animal model, experimental autoimmune neuritis (EAN).MethodsEAN in C57BL/6 mice induced by subcutaneous injection with peripheral nerve myelin P0 protein peptide 180–199 (P0 peptide) were treated with ginkgolides at three different doses. GBS patients were randomly divided into two groups, the experimental group and the control group. The experimental group were treated with ginkgolides as soon as diagnosed.ResultsOur data indicated that ginkgolides administration daily ameliorated the score of EAN and delayed the peak of disease in EAN mice. Ginkgolides also down-regulated the proportions of T helper (Th) 17 cells in EAN spleens. Furthermore, we also found that administration of ginkgolides significantly decreased the levels of interferon (IFN)-γ and interleukin-12 (IL)-12 in GBS patients.ConclusionsOur results suggested that ginkgolides ameliorated the clinical score of EAN through down-regulating the proportions of Th 17 cells. Ginkgolides also suppressed inflammation response by decreasing pro-inflammatory cytokines IFN-γ and IL-12, suggesting ginkgolides had potential therapeutic effects on GBS patients and EAN in the future.     

APOE ε3 attenuates experimental autoimmune neuritis by modulating T cell, macrophage and Schwann cell functions

Human apolipoprotein E (apoE) is a 34.2 kDa glycosylated protein with three isoforms (apoE2, apoE3 and apoE4). Experimental autoimmune neuritis (EAN), an animal model for human Guillain–Barré syndrome, is an immune-mediated experimental disorder of the peripheral nervous system (PNS). Increased susceptibility to EAN in apoE deficient mice has been previously found. To elucidate the isoform-dependent effects of apoE on EAN, we used human apoE2, E3 and E4 transgenic mice (Tg) immunized with P0 peptide 180–199, as well as T cell proliferation test, macrophage and Schwann cell (SC) cultures to investigate the effects of apoE isoforms on the functions of T cells, macrophages and SCs both under naïve conditions and in EAN. Clinical signs of EAN were most severe in wild type (WT) C57BL/6 mice and apoE4 Tg mice, followed by apoE2 Tg mice and apoE3 Tg mice (WT ≈ E4 > E2 > E3, p < 0.01). At the nadir of EAN, spleen weight and lymphocyte proliferation were in line with the clinical severity of the disease. Proliferation tests of purified T cells from naive mice stimulated with phytohemagglutinin or interleukin-12 showed isoform-specific differences (WT ≈ E4 > E3 ≈ E2, p < 0.01). Macrophages from both naïve and EAN mice produced nitric oxide upon inflammatory stimulation with lipopolysaccharide, interferon-γ, polyinosinic:polycytidylic acid or combinations thereof, in an isoform-dependent manner (WT ≈ E4 > E2 > E3, p < 0.01). Generalized intervention with 1400W, a specific inducible nitric oxide synthase inhibitor, significantly suppressed the clinical course of EAN in apoE2, E3 and E4 Tg mice and in WT mice. During the recovery stage of disease, the highest expression of CD178 (FasL) on SCs was found in apoE3 Tg mice. Our data support an isoform-dependent effect of apoE on EAN. This might be due to the isoform-specific effects of apoE on functions of T cells, macrophages and SCs, which contribute to the distinct clinical courses of EAN. ApoE3 might not only inhibit the onset and suppress the clinical severity of EAN, but also enhance the termination of immune responses in the PNS.     

糖皮质激素治疗多发性硬化不同方案的临床应用

目的探讨糖皮质激素治疗多发性硬化(MS)的不同方案的疗效、住院费用、住院时间、复发情况及不良反应等。方法 97例MS急性期患者随机分为鞘内注射甲泼尼龙(MPS)组(30例)、MPS冲击治疗组(36例)及地塞米松治疗组(31例),比较各组治疗后不同时间的Kurtzke扩展致残量表(EDSS)评分减少值、住院费用、费用中药品比例、住院时间、复发情况及不良反应等。结果鞘内注射MPS组治疗后5 d起,MPS冲击治疗组治疗后10 d起EDSS评分减少值明显高于地塞米松治疗组(均P<0.05)。治疗后5 d时鞘内注射MPS组EDSS评分减少值明显高于MPS冲击治疗组(P<0.05)。3组间鞘内注射MPS组总住院费用最少,费用中药品比例最低,日均住院费用较少,住院时间较短(均P<0.05);MPS冲击治疗组日均住院费用最高,地塞米松治疗组住院时间最长(均P<0.05)。MPS冲击治疗组复发率最高,复发时间最短(均P<0.05);地塞米松治疗组和鞘内注射MPS组复发率及复发时间比较差异无统计学意义。各组均无严重不良反应发生。结论鞘内注射MPS治疗MS起效较快,近期疗效显著,住院费用较低,复发率较低,复发时间较长,无严重不良反应,宜在临床推广应用。     

Hyperpolarisation-activated cyclic nucleotide channel 4 (HCN4) involvement in Tourette's syndrome autoimmunity

Previous studies have shown that interferon-gamma (IFN-γ) is a proinflammatory cytokine that contributes to the pathogenesis of Guillain-Barré syndrome and its animal model, experimental autoimmune neuritis (EAN). Treatments with anti-IFN-γ antibodies improve clinical outcome in GBS patients and EAN animals and administration of IFN-γ markedly worsens EAN. Paradoxically, the mice deficient in IFN-γ remain susceptible to experimental autoimmune encephalomyelitis, an analogous disease in the central nervous system. These observations raise a question whether IFN-γ might be protective in autoimmune demyelinating diseases. To clarify the role of IFN-γ in the pathogenesis of autoimmune demyelinating diseases, we used P0 protein peptide 180-199 to induce EAN in IFN-γ knockout (KO) mice. After the acute phase of EAN, the clinical signs of IFN-γ KO mice were significantly more severe than those of wild type (WT) controls. After antigenic stimulation, the proliferation of splenic mononuclear cells was significantly higher in IFN-γ KO than in WT mice with EAN. At the peak of EAN, the proportion of interleukin (IL)-17A expressing cells in cauda equina (CE) infiltrating cells, and the levels of IL-17A in sera were elevated in IFN-γ KO mice when compared with their WT counterparts. The proportions of major histocompatibility complex (MHC) II, macrosialin, and IL-12/IL-23p40 expressing cells, relative to total CE infiltrating cells were correspondingly higher in IFN-γ KO than in WT mice with EAN. However, IFN-γ deficiency reduced the production of NO by cultured macrophages in response to proinflammatory stimuli and induced a systemic Th2-oriented immune response. In conclusion, IFN-γ deficiency exacerbates EAN via upregulating Th17 cells despite a mitigated systemic Th1 immune response.     

黄芩苷对实验性自身免疫性脑脊髓炎大鼠髓鞘的保护作用

目的观察黄芩苷(BAC)对实验性自身免疫性脑脊髓炎(EAE)大鼠髓鞘的保护作用。方法将大鼠随机分为正常对照(NC)组、EAE组、地塞米松(DXM)组和BAC组。抗原免疫1周后分别予以DXM组和BAC组大鼠DXM和BAC治疗7d;观察免疫后14d各组动物发病情况、脊髓病理变化及髓鞘碱性蛋白(MBP)的表达。结果(1)EAE组、DXM组和BAC组大鼠于抗原免疫后8～10d发病,发病潜伏期分别为9.62d、11.0d、9.85d,DXM组较EAE组潜伏期显著延长(P<0.05),BAC组与EAE组间差异无统计学意义;各组发病率分别为75.0%、66.7%、80%,各组间差异无统计学意义。(2)病程中EAE组和DXM组质量较BAC组明显下降(均P<0.05);DXM组和BAC组神经功能评分明显优于EAE组(均P<0.05)。(3)与EAE组比较,DXM组脊髓病灶数明显减少(P<0.05),BAC组病灶数有所减少,但差异无统计学意义。(4)与EAE组比较,DXM组和BAC组脊髓MBP阳性数显著增多(均P<0.05)。结论BAC对缓解EAE大鼠的临床症状、减轻髓鞘脱失的作用与DXM相似,而没有质量降低的不良反应。     

CCR5 deficiency does not prevent P0 peptide 180-199 immunized mice from experimental autoimmune neuritis

Experimental autoimmune neuritis (EAN) is an inflammatory autoimmune demyelinating disease of peripheral nervous system (PNS) and represents an animal model of Guillain-Barré syndrome (GBS) in man. The inflammatory cell infiltrating into the PNS is a prerequisite for developing EAN. To explore the role of CC chemokine receptor 5 (CCR5) in the inflammatory process of EAN, we induced EAN in CCR5-deficient (CCR5(-/-)) mice with P0 protein peptide 180-199. We found that CCR5(-/-) mice showed a similar EAN clinical course and severity as well as profile of infiltrating macrophages and T cells in cauda equina (CE) of EAN and the same levels of spleen mononuclear cell (MNC) response to antigen and mitogen when compared with CCR5(+/+) control mice. However, increased IP-10 and MIP-1beta production in sciatic nerves were seen in CCR5(-/-) mice. These results suggest that CCR5 deficiency does not prevent P0 peptide 180-199-immunized mice from EAN. Increased MIP-1beta and IP-10 in sciatic nerves may compensate the CCR5 deficiency and contribute to inflammatory cells infiltrating to the PNS.     

髓鞘碱性蛋白对急性一氧化碳中毒大鼠最终结局的预测价值及地塞米松干预的作用

目的评估髓鞘碱性蛋白(MBP)对急性一氧化碳(CO)中毒大鼠最终结局的预测价值,并探讨不同剂量地塞米松对急性CO中毒大鼠最终结局的干预作用。方法将130只体质量180²80g、雄性Wistar大鼠随机分成3个实验组(每组n=40)和健康对照组(n=10),即CO中毒组(CO中毒组);CO中毒+10mg·Kg-1·d-1地塞米松组(DXM-10组);CO中毒+30mg·Kg-1·d-1地塞米松组(DXM-30组)和健康对照组(NC组)。观察各组大鼠在染毒后21d内的死亡例数,并在染毒后60min断尾取血检测各组大鼠血清中MBP值。结果中毒后所有大鼠呈现典型急性CO中毒表现。在观察的21d内,CO中毒组中共有15只大鼠死亡,DXM-10组11只,而DXM-30组4只,死亡率分别为37.50%、27.50%和10.00%。而NC组中无大鼠死亡。实验组中所有死亡大鼠与存活大鼠其平均MBP值相比差异有统计学意义(P<0.05)。结论 CO中毒后60min腹腔内注射10mg·Kg-1·d-1地塞米松可降低急性CO中毒大鼠的死亡率,30mg·Kg-1·d-1地塞米松可显著降低其死亡率。MBP对急性CO中毒大鼠的最终结局具有预测价值。     

Aggravation of experimental autoimmune neuritis in TNF-alpha receptor 1 deficient mice

The role of tumor necrosis factor (TNF)-alpha and its receptors in the pathogenesis of experimental autoimmune neuritis (EAN) induced by P0 peptide 180-199 in TNFR1 (p55) deficient (TNFR1-/-) mice was investigated. Compared to wild type EAN mice, TNFR1-/- EAN mice developed significantly more severe clinical signs, in parallel with enhanced numbers of inflammatory infiltrating cells in peripheral nerves and splenic P0-reactive T cell proliferation, as well as increased obviously MHC class II and CCR3 expression on the macrophages in the cauda equina. Our data indicated that TNF-alpha might have anti-inflammatory effect preventing the development of EAN in this mouse model.