实验性自身免疫性脑脊髓炎大鼠脑组织中NF-κB p65,TNF-α表达及雷公藤多甙的干预研究

目的 观察Wistar大鼠实验性自身免疫性脑脊髓炎(EAE)脑组织中核转录因子-κBp65(NF-κB p65)、肿瘤坏死因子-α(TNF-a)的表达情况,用雷公藤多甙(TWP)进行干预,探讨TWP对EAE的防治效果及作用机制.方法 建立Wistar大鼠EAE模型,并选用TWP、泼尼松(PRD)干预治疗.65只Wistar大鼠随机分成5组:对照组5只,EAE组、TWPⅠ组(EAE+TWP 10 mg·kg-1·d-1)、TWPⅡ组(EAE+TWP 30mg·kg-1·d-1)和PRD组(EAE+PRD 5 mg·kg-1·d-1)各15只.观察各组大鼠发病情况并进行神经功能评分.大鼠脑和脊髓腰膨大段组织切片:苏木素-伊红(HE)染色+罗克沙尔坚牢蓝(LFB)髓鞘染色,光镜下进行炎性病灶计数,并免疫组化方法检测大鼠脑组织内NF-κB p65、TNF-α的表达.结果 EAE组及干预组大鼠脑组织内NF-κB p65、TNF-α表达水平随病情加重而升高,随病情缓解而降低.与EAE组比较,干预组大鼠发病率减低,潜伏期延长,神经功能评分及体重损失明显降低,脑组织炎性病灶数明显减少;同时脑组织NF-κB p65和TNF-α阳性细胞数量明显减少.TWP大剂量组与PRD治疗效果相当,且优于TWP小剂量组.结论 NF-κB p65、TNF-α过表达可能参与EAE发生及进展;TWP有明显防治大鼠EAE作用,在一定程度上呈剂量相关性,其机制可能与下调NF-κB p65、TNF-α表达有关.     

核转录因子-κB在糖尿病大鼠穹窿下器细胞的表达

目的观察核转录因子-κB(NF-κB)在不同时期糖尿病大鼠穹窿下器(SFO)中的表达并探讨其表达的意义。方法雄性Wistar大鼠,模型组每只大鼠给予链脲佐菌素60mg·kg-1,腹腔一次性注射,建立1型糖尿病大鼠模型。用免疫组织化学染色方法观察NF-κB在正常大鼠及2、4、8 w糖尿病大鼠SFO细胞内的表达。结果对照组NF-κB免疫阳性细胞稀少,胞核呈浅棕黄色,平均吸光度为156.42±7.25。2w组NF-κB免疫强阳性细胞SFO内均匀分布,核呈棕褐色,高度表达,为强阳性,平均吸光度为183.72±8.35。4w组SFO NF-κB的表达水平有所下降但仍高于正常对照组,平均吸光度为169.55±5.84。8w组NF-κB免疫强阳性细胞的表达与正常对照组无明显区别,平均吸光度为158.32±8.72。平均吸光度测定:2w组>4w组>8w组,P﹤0.05,8w组与对照组比较P﹥0.05。结论 NF-κB在糖尿病大鼠SFO细胞内呈短暂一过性的表达增强,NF-κB途经可能在糖尿病大鼠SFO细胞损伤中起着重要的作用。     

rhEPO对大鼠颅脑损伤后的损伤脑组织NF-κB表达和血清TNF-α水平的影响

目的探讨重组人促红细胞生成素（rhEPO）对大鼠颅脑损伤后损伤脑组织核因子-κB（NF-κB）表达和血清肿瘤坏死因子-α（TNF-α）水平的影响。方法将90只雄性成年Wistar大鼠分为假损伤组、颅脑损伤组及rhEPO组。采用改进Feeney法制作大鼠颅脑损伤模型,rhEPO组伤后即刻腹腔注射rhEPO（3000IU／kg）;伤后3h、12h、24h、48h、72h、5d和7d免疫组织化学染色法检测NF-κB的表达、双抗体夹心酶联免疫吸附法检测血清TNF-α水平。结果伤后3h,损伤脑组织NF-κB表达明显升高,伤后24h达最高峰,随后其表达水平逐渐下降,至伤后7d仍明显升高（P〈0．05）;rhEPO治疗后,每个时间点损伤脑组织NF-κB表达水平均明显降低（P〈0.05）。伤后3h,血清TNF-α水平也明显升高,伤后12h迅速达到最高峰,随后其水平逐渐下降;伤后3d,其水平又再次升高,随后又逐渐下降,至伤后7d仍明显升高（P〈0．05）。rhEPO治疗后,每个时间点血清TNF-α水平均明显降低（P〈0．05）。结论NF-κB和TNF-α可能在颅脑损伤后的炎症反应中发挥重要作用,而rhEPO可能通过抑制NF-κB和TNF-α的表达而减少损伤脑组织的炎症反应,发挥脑保护作用。     

大鼠脑出血后核因子-κB的表达及黄芪多糖的干预作用

目的研究黄芪多糖（astragalus polysaccharide，APS）对大鼠脑出血（ICH）后血肿周围核因子-κB（nuclear factor-kappa B，NF-κB）蛋白表达的影响，并探讨APS抗炎的脑保护机制。方法采用Ⅶ型胶原酶注射至大鼠右侧苍白球诱导ICH模型并于造模术后2h行APS腹腔注射，分别于6h和1、3、5d4个时间点进行大鼠神经行为学评分，采用免疫组化方法检测血肿周围组织NF-κB表达的动态变化，采用透射电镜观察术后3d血肿周围神经元超微结构的变化。结果与模型组比较，APS干预后3d和5d大鼠神经行为学评分明显减少（P〈0．05），干预6h及1、3、5d时NF-κB阳性细胞数明显减少（P〈0．05），神经元超微结构改变亦比模型组减轻。结论APS可抑制NF—κB激活，减轻炎症反应，改善神经功能缺损症状。     

Nrf2-ARE信号通路对脑出血灶周神经保护作用的机制研究

目的研究大鼠脑出血后Nrf2-ARE通路HO-1对出血灶周神经保护作用及其相关机制探讨。方法采用SD大鼠基底节自体股动脉血注射法建立脑出血模型,分4组:单纯脑出血(ICH)组、莱菔硫烷(SFN)组、维甲酸(RA)组、对照组,观察不同时间点大鼠神经功能评分,取脑组织行免疫荧光检查、Western blot、RT-PCR检测Nrf2、HO-1、NF-κB、TNF-α表达。对所得数据进行统计分析,观察这些指标的变化。结果 SFN组与ICH组比较,神经功能障碍明显减轻,出血灶周Nrf2因子、HO-1抗氧化蛋白表达均升高,于3 d达高峰值,持续表达至7 d;SFN组NF-κB、TNF-α表达下降;RA组大鼠死亡率最高,神经功能障碍较ICH组及SFN组均严重,Nrf2因子、HO-1抗氧化蛋白表达均较SFN组和ICH组低,持续性抑制作用可能持续7 d左右,RA处理组NF-κB、TNF-α表达升高,炎性反应持续时间可能超过7 d。结论 RA抑制Nrf2解离及转入核,抑制Nrf2-ARE信号通路的抗炎性作用,SFN激活Nrf2-ARE信号通路可提高HO-1抗氧化酶表达,减轻脑出血后灶周炎性反应,具有神经保护作用,提示Nrf2-ARE信号通路将是治疗脑出血灶周炎性损伤的新方向。     

黄芪多糖对脑出血大鼠脑组织含铁血红素氧合酶-1蛋白表达及含水量、超微结构的影响

目的 研究黄芪多糖对脑出血大鼠脑组织含铁血红素氧合酶-1(HO-1)蛋白表达及含水量、超微结构的影响.方法 139只雄性SD大鼠随机分为假手术组、脑出血组与黄芪多糖治疗组.采用Ⅶ型胶原酶诱导法制作SD大鼠脑出血模型,假手术组以等量生理盐水代替Ⅶ型胶原酶;黄芪多糖治疗组给予黄芪多糖25 mg/kg腹腔注射,每天1次,直至处死.分别于脑出血后12 h、24 h、2 d、4 d、7 d 5个时间点对各组大鼠进行神经行为学评分,测量脑组织含水量和用免疫组化法检测HO-1蛋白表达,透射电镜观察脑出血后4 d 血肿周围神经元的超微结构.结果 脑出血组与黄芪多糖治疗组大鼠脑出血后脑组织 HO-1 表达以及脑组织含水量较假手术组显著增高(均P＜0. 01);脑出血后12 h HO-1蛋白即有表达, 2 d 达高峰, 尔后逐渐下降. 黄芪多糖治疗组与脑出血组比较, HO-1表达的变化相同,但各时间点表达均高于脑出血组(P＜0.05～0.01);脑组织含水量各时间点均显著降低(P＜0.05～0.01);4 d、7 d时大鼠神经行为学评分明显减少(均P＜0.01),神经元超微结构病理改变轻.结论 黄芪多糖可促进脑出血后脑组织HO-1的表达, 这可能是其减轻脑出血后脑水肿和脑组织损伤的机制之一.     

雌激素抑制实验性自身免疫性脑脊髓炎大鼠的NF-κB和Rho激酶表达

目的 探讨雌激素对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)大鼠中枢神经系统NF-κB和Rho激酶表达的影响及相关作用机制.方法 60只雌性Wistar大鼠行卵巢摘除术后建立EAE模型,随机分为模型组、对照组、雌激素组和NF-κB特异性抑制剂吡咯烷二硫代氨基甲酸盐(pyrro-lidine dithocarbamate,PDTC)组,每组15 只.自免疫日起,雌激素组给予17β-雌二醇20 μg/(kg·d),连续10 d;PDTC组注射PDTC 50 mg/(kg·d),连续10 d.免疫后16天统一处死取脑组织,HE染色观察炎性细胞浸润情况,免疫组化和Western blot 观察NF-κB p65和Rho激酶表达的变化.结果 与模型组比较,雌激素组发病率无统计学差异(P>0.05),但发病潜伏期延长(P<0.05),神经功能评分较低(P<0.05);HE染色示雌激素组和PDTC组脑组织炎性细胞浸润程度较模型组轻;免疫组化和Western blot 结果示,与模型组相比,雌激素组和PDTC 组NF-κB和Rho激酶表达均较低(P<0.01).结论 雌激素能抑制EAE大鼠NF-κB和Rho激酶的表达,其对Rho激酶的抑制作用可能部分通过下调NF-κB的水平实现的.     

异氟烷预处理对偏头痛大鼠行为学症状及三叉神经节相关蛋白表达的影响

目的探讨异氟烷(ISO)对硝酸甘油所致偏头痛大鼠行为学症状及三叉神经节内相关蛋白表达的影响。方法采用随机数字表法将SD大鼠分为对照组(S组)、模型组(M组)、ISO预处理组(包括低剂量和高剂量组,即L组和H组),每组10只,并采用皮下注射硝酸甘油法制备偏头痛模型。L组和H组于造模前30 min分别给予1%和2%ISO吸入麻醉30 min。观察并比较各组大鼠造模后耳红出现和消失的时间及每30 min时间(T_(1-7))段内的挠头、爬笼次数。采用Western blot测定三叉神经节内白介素-1β(IL-1β)、环氧合酶-2(COX2)、降钙素基因相关肽(CGRP)及核转录因子-κB p65(NF-κB p65)的蛋白表达水平。结果与S组相比,M组和ISO预处理组大鼠造模后均出现双耳发红及挠头、爬笼次数均增多等现象,三叉神经节内IL-1β、COX-2、CGRP及胞核NF-κB p65蛋白表达水平明显升高,胞浆NF-κB p65蛋白表达水平明显降低(P 0. 05);与M组相比,ISO预处理组大鼠耳红消失的时间明显缩短,T_(2-7)时挠头次数均明显减少,T_(2-6)时爬笼次数均明显减少,三叉神经节内IL-1β、COX-2、CGRP及胞核NF-κB p65蛋白表达水平明显降低,胞浆NF-κB p65蛋白表达水平明显升高(P 0. 05),且H组较L组更明显(除胞浆NF-κB p65蛋白外)。结论异氟烷能改善偏头痛大鼠的行为学症状,其机制可能与下调三叉神经节内IL-1β、COX-2、CGRP蛋白的表达水平及抑制NF-κB的激活有关。     

脑心通对缺血性大鼠脑组织核转录因子-κB、基质金属蛋白酶-9和肿瘤坏死因子-α的影响

目的探讨脑心通对缺血性大鼠脑组织核转录因子-κB(NF-κB)、基质金属蛋白酶-9(MMP-9)和TNF-α的影响。方法将110只健康雄性SD大鼠随机分为假手术组(30只),模型对照组(40只),脑心通治疗组(40只),每组再分为缺血再灌注后12 h、1 d、3 d、5 d、7 d五个时间点进行处理。采用Zea Longa法制备大脑中动脉闭塞缺血再灌注模型,利用MRI检查和Bederson评分筛选成功模型;每组五个时间点分别选取6只大鼠行MRI检查,动态观察使用脑心通胶囊后缺血脑组织变化过程,采用western blot、实时定量PCR分别检测梗死区脑组织NF-κB、MMP-9和TNF-α的蛋白和mRNA表达水平。结果脑心通治疗组脑梗死区域体积较模型对照组减少。脑心通治疗组NF-κB、MMP-9、TNF-α蛋白及其mRNA水平均明显低于模型对照组(均P0.05);假手术组MMP-9、TNF-α和NF-κB蛋白及其mRNA水平与脑心通治疗组差异无统计学意义。结论脑心通通过改变脑梗死区周围炎症因子表达水平来降低炎症反应,发挥神经保护作用。     

β-七叶皂甙钠对大鼠局灶性脑缺血再灌注后NF-κB、ICAM-1、VCAM-1表达的影响

目的 探讨大鼠局灶性脑缺血再灌注脑组织缺血区不同时间点NF-κB、ICAM-1、VCAM-1蛋白表达的变化,及β-七叶皂甙钠干预效果.方法 采用大鼠大脑中动脉闭塞法(MCAO)制作局灶性脑缺血再灌注模型,用免疫组织化学方法观察大鼠脑缺血再灌注不同时间段,NF-κB、ICAM-1、VCAM-1蛋白的表达.并在大鼠于脑缺血前24h、1h及再灌注即刻分别腹腔给予β-七叶皂甙钠5mg/kg,2h MCAO,再灌注24h、48h后取脑,运用TTC染色测算脑梗死体积,免疫组化染色检测NF-κB、ICAM-1、VCAM-1蛋白表达,分析β-七叶皂甙钠的干预效应.结果 (1)脑缺血后缺血区脑组织NF-κB及ICAM-1、VCAM-1表达均增加,NF-κB于再灌注后12～24h表达达高峰,ICAM-1于再灌注后24h表达达高峰,VCAM-1于再灌注后24～48h表达达高峰.(2)NF-κB的表达与血管内皮ICAM-1、VCAM-1的表达呈正相关.(3)β-七叶皂甙钠能显著降低脑缺血再灌注后24h和48h缺血区NF-κB、ICAM-1及VCAM-1的表达增加.(4)β-七叶皂甙钠能明显减轻脑缺血再灌注后的脑组织损伤,再灌注24h脑梗死体积减少41.8%.结论 (1)脑缺血再灌注后NF-κB、ICAM-1、VCAM-1大量表达,这可能是脑缺血再灌注损伤机制之一.(2)脑缺血后NF-κB的活化可能与微血管内皮细胞ICAM-1、VCAM-1蛋白表达调控有关.(3)β-七叶皂甙钠能够减轻脑缺血后的脑组织损伤,有神经保护作用.     

Lesional accumulation of RhoA(+) cells in brains of experimental autoimmune encephalomyelitis and multiple sclerosis

Aim: Infiltration of autoantigen-specific T cells and monocytes into the central nervous system is essential for the development of both experimental autoimmune encephalomyelitis (EAE) and multiple sclerosis (MS). RhoA is one of the best-known members of Rho GTPases, and inhibition of RhoA has been shown to attenuate the progression of EAE. The aim of this study was to investigate the expression of RhoA in brains of EAE rats and MS tissue. Methods: EAE was induced by immunization with the synthetic peptide gpMBP68-84 in rats, and clinical severity was scored. RhoA expression pattern was investigated in brains of EAE rats at different time points and in different lesions of brain tissue specimens from six MS brains and five neuropathologically unaffected controls by immunohistochemistry. Methods: In EAE rat brains, accumulation of RhoA⁺ cells reached maximal levels around Day 13, correlating to the clinical severity of EAE, and up-regulation lasted until the recovery stage of the disease. Double-labelling experiments showed that the major cellular sources of RhoA were reactive macrophages/microglia. While RhoA⁺ cells in normal human brain parenchyma were rarely observed, RhoA expression was found to be spatially associated with MS lesions, showing a marked decrease from active lesions via chronic stages to its near absence in normal-appearing white matter. In addition, major RhoA⁺ cells in brain parenchyma of MS were identified to be activated macrophages/microglia. Conclusion: Our present data indicated that RhoA may play an important role during the effector phase of EAE and MS. Therefore, RhoA inhibitors might be a therapeutic option for MS patients.     

Expression of citrullinated proteins in murine experimental autoimmune encephalomyelitis

In this study, we demonstrate for the first time the immunohistochemical expression of citrullinated proteins in the central nervous system (CNS) of mice with myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE). By using an established monoclonal antibody (F95) against natural and synthetic citrullinated proteins (Nicholas and Whitaker [2002] Glia 37:328-336), numerous, small, previously unrecognized "patches" of citrullinated proteins were discovered throughout EAE brains, whereas EAE spinal cords showed similar but much larger lesions. On dual color immunofluorescence, these lesions were found to contain citrullinated myelin basic protein (MBP) and were surrounded by astrocytes immunoreactive for both glial fibrillary acidic protein (GFAP) and F95. These lesions became evident about the time when EAE mice became symptomatic and increased in size and number with increasing disease severity. In some sections of spinal cord but not brains of severely debilitated EAE mice, a widespread gliotic response was seen, with astrocytes containing citrullinated GFAP spread throughout the gray and white matter. Western blot analysis of acidic proteins from the brains and spinal cords of EAE mice had higher levels of multiple citrullinated GFAP isoforms compared with controls, with more F95-positive bands in the EAE brains vs. spinal cords. These results raise the possibility that citrullination of both GFAP and MBP may contribute to the pathophysiology of EAE and that the brains of EAE mice may contain more pathology than previously realized.     

Heme oxygenase-1 plays an important protective role in experimental autoimmune encephalomyelitis.

Increasing evidence shows that oxidative stress plays an important role in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), an animal model of the human disease, multiple sclerosis (MS). Heme oxygenase-1 (HO-1) is a heat shock protein induced by oxidative stress. HO-1 metabolizes heme to the antioxidant bilirubin and carbon monoxide, and represents a powerful endogenous defensive mechanism against free radicals in many diseases. However, the role of this important enzyme in EAE remains unknown. In this study, we showed high expression of HO-1 in lesions of EAE, and demonstrated that hemin, an inducer of HO-1, inhibited EAE effectively. In contrast, tin mesoporphyrin, an inhibitor of HO-1, markedly exacerbated EAE. Our results suggest that endogenous HO-1 plays an important protective role in EAE, and that targeted induction of HO-1 overexpression may represent a new therapy for the treatment of multiple sclerosis.     

Differential upregulation of heme oxygenase-1 (HSP32) in glial cells after oxidative stress and in demyelinating disorders

Oxidative stress is implicated in the pathogenesis of demyelinating disorders and inflammatory responses. Heme oxygenase-1 (HO-1; HSP32) is a small heat shock protein (HSP) with enzymatic activity, which is inducible by oxidative stress. In this study we analyzed autopsy and biopsy brain samples of patients with multiple sclerosis (MS) and ADEM (acute disseminated leucoencephalomyelits) and spinal cord lesions of mouse EAE (experimental autoimmune encephalomyelitis), which was actively induced by immunization with myelin oligodendrocyte glycoprotein (MOG_35–55) peptide, for the presence of HO-1. HO-1 was observed in glial cells during different stages: (1) during acute phases of mainly inflammatory diseases (EAE and ADEM) expression of HO-1 was prominent in microglia/macrophages and astrocytes, and upregulation correlated with inflammation, and (2) in early MS lesions HO-1 was expressed in oligodendrocytes. Furthermore, in glial cell cultures, we can show that upregulation of HO-1 in oligodendrocytes was paralleled by severe morphological damage. Oligodendrocytes underwent apoptotic cell death at a concentration of hydrogen peroxide (50–200 μM) which did not affect astrocytes or microglia. Using oligodendroglial OLN-93 cells, we demonstrate that oxidative stress led to mitochondrial impairment and the disorganization of the microtubule network. Zinc protoporphyrin, an inhibitor of HO-1, augmented the cytotoxic consequences of hydrogen peroxide in OLN-93 cells. Hence, the presence of HO-1 in EAE, ADEM, and MS points to the involvement of oxidative stress and a role of HO-1 in the pathogenesis of the diseases. The data suggest that stress-induced HO-1 initially plays a protective role, while its chronic upregulation, might contribute to oligodendroglial cell death rather than providing protection.     

NF-KB p65及环氧化酶-2在实验性变态反应性脑脊髓炎中的表达及其作用机制

目的 研究核转录因子NF-kB p65及环氧化酶-2(COX-2)在实验性变态反应性脑脊髓炎(EAE)中的表达,探讨其参与EAE的作用机制. 方法 健康新西兰兔30只根据采用不同免疫原制作模型的方式分为三组:A组,正常对照组(未注射免疫原)10只;B组,新西兰兔脊髓匀浆(R-SCH)EAE模型组,10只;C组,牛脑匀浆(C-SCH)EAE模型组,10只.比较每组动物临床病理特点;分别采用免疫荧光、免疫组化对其进行形态观察,应用ELISA法对NF-KBp65进行活性测定.结果 B、C组均可见NF-kB p65及COX-2明显增加,阳性细胞百分率与A组比较.差异有统计学意义(P<0.05);其中B组NF-kB p65活性明显增高.结论 在EAE模型中存在NF-kBp65及COX-2表达,它们均在EAE发病过程中发挥了重要作用.     

神经妥乐平通过Nrf2/HO-1/NQO1通路发挥对帕金森病大鼠的神经保护作用

目的 探讨神经妥乐平对帕金森病(PD)大鼠的神经保护作用及其相关机制.方法 SD大鼠分为对照组、PD组(PD造模)、神经妥乐平低剂量组(PD造模+腹腔注射0.6 Nu·kg-1神经妥乐平溶液)、神经妥乐平高剂量组(PD造模+腹腔注射1.2 Nu·kg-1 神经妥乐平溶液)和通路抑制组(PD造模+腹腔注射1.2 Nu·k...     

Expression of allograft inflammatory factor-1 and haeme oxygenase-1 in brains of rats infected with the neurotropic Borna disease virus

Experimental infection of Lewis rats with Borna disease virus (BDV) causes an immune-mediated nonpurulent meningoencephalitis. Viral persistence in the central nervous system is accompanied by mononuclear infiltrates, activated monocytic/microglial cells and reactive astrocytes. The immune-mediated process was further characterized by expression analysis of allograft inflammatory factor-1 (AIF-1), a novel marker of monocyte/microglial activation and of glial fibrillary acid protein (GFAP) between day 3 and day 50 post infection (p.i.). Potential neuroprotective effects of these cells were studied by the induction of haeme oxygenase-1 (HO-1), a defensive molecule against oxidative stress in various brain insults. In BDV-infected rat brains, mononuclear infiltrates and AIF-1 expression increased up to day 28 p.i. During early time points p.i., AIF-1 expression was mainly found in inflammatory lesions and adjacent brain parenchyma. Already 24 days p.i., a widespread upregulation of AIF-1 was observed which declined only moderately beyond day 28 p.i. HO-1 induction was maximal between days 18 and 28 p.i. Increased amounts of GFAP-positive astrocytes were present beyond 24 days p.i. Viral antigen expression increased simultaneously to the inflammatory reaction and persisted up to 50 days p.i. Widespread upregulation of AIF-1 indicates an early, long-lasting microglial activation, which might be involved in the immunesurveillance of the immune-mediated inflammatory events. The early peak of HO-1 most likely represents a neuroprotective, anti-inflammatory response by invading monocytes, microglial cells and astrocytes during the formation of encephalitic lesions and acute viral replication.     

实验性变态反应性脑脊髓炎大鼠穹窿下器凋亡细胞的观察

目的通过观察实验性变态反应性脑脊髓炎(EAE)大鼠不同时期下穹窿下器(SFO)凋亡细胞的变化,进一步探讨该室周器官在EAE发病过程中的意义。方法建立大鼠EAE模型,利用TUNEL技术来观察大鼠SFO在EAE不同时期细胞凋亡的改变。结果SFO的凋亡细胞在EAE 6小时、EAE 7天、EAE 14天、EAE 21天分别为(13.67±2.31)、(19.33±2.08)、(24.67±6.79)、(9.33±1.76),而对照组为(5.33±2.52)。与对照组比较,EAE 6小时、7天、14天组细胞凋亡数明显增加(P<0.05),EAE 21天组无显著差异(P>0.05)。结论SFO在EAE的不同时期凋亡细胞的变化与临床进展呈一致性,可能是血携免疫分子早期入脑的位点。     

盐酸法舒地尔治疗实验性自身免疫性脑脊髓炎的潜能与抗炎作用

目的：探讨盐酸法舒地尔对实验性自身免疫性脑脊髓炎（EAE）的治疗效果及机制。方法：雌性C57BL/6小鼠,随机分为EAE对照组、盐酸法舒地尔干预组和盐酸法舒地尔治疗组。采用髓鞘少突胶质细胞糖蛋白多肽诱导慢性EAE模型。干预和治疗分别在免疫后第3天和症状出现时予以腹腔注射盐酸法舒地尔,观察EAE模型小鼠体重变化和临床症状,进行苏木精-伊红和CD4＋T细胞染色,同时检测磷酸化肌球蛋白磷酸酶（p-MYPT1）和核因子（NF-κB）。结果：盐酸法舒地尔可推迟并改善EAE小鼠症状,减轻中枢神经系统炎细胞浸润,抑制脊髓和脑p-MYPT1及脊髓NF-κB的表达。     

缺氧预处理对颅脑损伤大鼠脑组织HIF-1α及其HO-1表达的影响

目的探讨缺氧预处理对颅脑损伤大鼠脑组织缺氧诱导因子（HIF-1α）及血红素氧合酶-1（HO-1）表达的影响。方法 Sprague-Dawley大鼠102只,随机分为对照组（n=6）、创伤组（n=48）和预处理组（n=48）。创伤组按照改进的Feeney自由落体撞击法建立大鼠颅脑损伤模型,预处理组给予缺氧预处理后,同法造模。采用RT-PCR和Western blotting观察伤后1 h、4 h、8 h、12 h和1 d、3 d、7 d、14 d挫伤周围脑组织HIF-1α、HO-1表达变化。结果创伤组与对照组比较,HIF-1α和HO-1在伤后4 h、8 h、12 h和1、3 d表达上调（P〈0.05）。预处理组与创伤组比较,HIF-1α和HO-1在伤后1 h逐渐上调,伤后4 h、8 h、12 h和1、3 d表达显著上调,直至伤后7 d（P〈0.05）。结论缺氧预处理可增加颅脑损伤后挫伤区周围脑组织HIF-1α的表达,进而促进HO-1 m RNA及蛋白表达。其机制可能是缺氧预处理提高颅脑损伤对缺氧的适应性,减轻挫伤周围脑组织氧自由基对神经细胞伤害。