神经节苷酯对大鼠脑缺血神经细胞凋亡的影响

目的研究神经节苷脂(GM)对大鼠脑缺血神经细胞凋亡的抑制作用及可能机制。方法线栓法制作大鼠大脑中动脉栓塞模型,♂SD大鼠,随机分成假手术组、模型对照组、GM低、中、高剂量治疗组,缺血48 h后各组大鼠神经功能缺陷评分,测定脑组织一氧化氮(NO)的含量,检测脑细胞凋亡率,分析脑缺血区诱生型一氧化氮合酶(iNOS)和Caspase-3 mRNA的表达。结果与假手术组比较,模型对照组、GM低、中、高剂量治疗组神经功能缺陷评分、NO含量、凋亡率、iNOS和Caspase-3表达量均增加,差异有统计学意义(P<0.05);与模型对照组和GM低剂量治疗组比较,GM中、高剂量治疗组均明显下降,差异有统计学意义(P<0.05);模型对照组和GM低剂量治疗组之间差异无统计学意义(P>0.05),GM中剂量治疗组和高剂量治疗组之间差异也无统计学意义(P>0.05)。结论神经节苷酯减少iN-OS生成,抑制细胞凋亡,对缺血神经细胞产生保护作用。     

GD1a对基质金属蛋白酶-9在不同细胞类型中表达的调控作用

目的在不同的细胞类型中,检测神经节苷脂GD1a是否抑制基质金属蛋白酶-9(MMP-9)的表达。方法采用逆转录PCR法和酶谱法检测MMP-9及MMP-2的表达。结果在鼠黑色素瘤B16细胞中,神经节苷脂GD1a促进MMP-9的表达,而在小鼠肺癌Lewis细胞、人单核细胞THP-1及人子宫颈癌HeLa细胞中,神经节苷脂GD1a降低MMP-9的表达。在上述细胞中,神经节苷脂GD1a不影响基质金属蛋白酶MMP-2的表达;肿瘤坏死因子TNF-α在这几种细胞中均能促进MMP-9的表达,但对MMP-2无影响;在B16细胞中,GD1a通过胞窖膜介导信号传递,并且主要通过ERK促进MMP-9的表达,而在Lewis细胞中,GD1a不通过胞窖膜介导信号传递,但部分通过p38、ERK和JNK抑制MMP-9表达。结论在不同的细胞类型中,神经节苷脂GD1a能够激活不同的信号转导途径。     

单唾液酸神经节苷脂对体外循环后大鼠脑组织氧化性应激的影响

目的：探讨单唾液酸神经节苷脂（GM1）对大鼠体外循环（CPB）后脑组织的氧化应激反应的变化及GM1干预后对其的影响。方法：选用雄性SD大鼠,经右颈静脉插管引流,尾动脉插管灌注建立CPB,转流时间1 h,建立CPB动物模型。随机分为CPB模型组、CPB模型＋GM1组、输血组和假手术组,每组10只。脑组织匀浆测定乳酸脱氢酶、丙二醛、超氧化物歧化酶和谷胱甘肽过氧化物酶等酶的活性。结果：CPB后脑组织中,乳酸脱氢酶由正常的（38.21±3.61）U/mg增加到（62.17±6.71）U/mg（P〈0.01）,给予GM1后下降到（51.63±5.92）U/mg（P〈0.05）,与CPB组比较有显著差异（P〈0.05）;MDA由正常的（2.14±0.81）nmol/mg升高到（6.76±1.31）nmol/mg（P〈0.01）,给予GM1后下降到（4.53±0.93）nmol/mg（P〈0.05）,与CPB组比较有显著差异（P〈0.05）;而SOD、GSH-Px的活性分别由正常的（1173.72±268.75）mmol/h·mg^-1、（128.32±42.74）mmol/h·mg^-1降低到（586.74±133.46）mmol/h·mg^-1、（73.19±18.67）mmol/h·mg^-1（P〈0.01）,给予GM1后分别回升到（723.19±189.65）mmol/h·mg^-1、（89.41±22.16）mmol/h·mg^-1,与CPB组比较有显著差异（P〈0.05）。结论：提示CPB脑损伤的过程中氧自由基是重要的致病因素,GM1可以明显减少氧自由基的产生和促进SOD、GSH-Px的产生。     

神经生长因子和神经节苷脂GM1联合应用对鼠基底前脑胆碱能神经元的保护作用

目的 探讨神经生长因子(NGF)和神经节苷脂GM1联合应用对基底前脑胆碱能神经元是否具有延缓退变的作用.方法 实验分为NGF组、NGF+神经节苷脂GM1组和单纯对照组.取孕20 d SD大鼠胚基底前脑原基制成细胞悬液接种子24孔培养板中,按分组加入NGF、NGF+神经节苷脂GMI和不含神经营养因子的DMEM培养液,分别于体外培养12、18、24、30 d后进行乙酰胆碱酯酶组织化学反应.显微镜下计数各孔中乙酰胆碱酯酶阳性神经元数,每孔随机测量和计数20个乙酰胆碱酯酶阳性神经元的平均胞体直径、发出的突起数和最长突起长度.数据用方差分析和SNK检验进行统计学处理.结果 在培养的4个时期,NGF组和联合组的各项数据均明显优于单纯对照组,神经节苷脂GM1+NGF组的各项数据,特别是最长突起长度优于单独使用NGF组.结论 NGF和神经节苷脂GM1联合应用及NGF单独应用不仅对体外培养的胚胆碱能神经元发育生长具有促进作用,而且还可延缓体外长期培养的人鼠胚基底前脑胆碱能神经元的退变;联合应用效果更佳.     

单唾液酸四己糖神经节苷脂钠注射液不良事件及其救护措施

目的探讨单唾液酸四己糖神经节苷脂钠注射液( GM1)致药品不良事件( ADE)的相关因素及其救护措施,为临床合理用药提供依据。方法对湖南中医药大学第一附属医院 2019年 1月至 2020年 12月发生的 12例 GM1致 ADE报告进行回顾性分析。结果 GM1致 ADE多在用药后 5 min~12 d内,主要表现为寒战、发热等全身性损害 7例次;其余为皮肤损害 3例次、呼吸系统与心血管系统损害各 2例次、神经系统损害 1例次,以及出现说明书未提及的视物模糊 1例次,经过停药并对症处理后上述症状均明显缓解。结论临床在使用 GM1前应详细询问病人过敏史,优先选择 5%葡萄糖注射液做溶媒,缓慢滴注;在用药过程中应加强 ADE监测,一旦发生立即停药,并考虑使用地塞米松注射剂 5 mg或 10 mg静滴等对症治疗。     

Retinal cholinergic and dopaminergic deficits of aged rats are improved following treatment with GM1 ganglioside

Selected cholinergic and dopaminergic markers were compared in the retina of aged (20–22-months-old) and young (3-months-old) rats before and after treatment with GM1 ganglioside. The dopaminergic markers, tyrosine hydroxylase, aromatic L-amino acid decarboxylase, dopamine, 3,4-dihydroxyphenylacetic acid and homovanillic acid were comparable in the young and aged animals and GM1 treatment did not alter them. In contrast, mazindol binding, a marker for the dopamine transporter, was diminished in the aged retina and treatment with GM1 restored binding to values found in the young animals. The cholinergic markers choline acetyltransferase and hemicholinium-3 binding, a marker for the high-affinity choline transport, were depressed in aged rats and GM1 corrected the deficits.     

抗神经节苷脂GM1抗体对重症多神经病变患者的诊断价值

目的对重症多神经病变（CIP）高危人群血清抗GM1 IgG抗体的测定，探讨其对CIP的诊断价值。方法以综合ICU收治的机械通气时间≥7d的患者于第7天行肌电图检查，按检查结果将患者分为CIP组和非CIP组。肌电图检查当日抽取空腹静脉血，测定抗GM1 IgG抗体，比较两组间的差异，并分析与肌电图神经损伤的关系。结果CIP组抗GM1 IgG抗体水平高于非CIP组，两组差异有统计学意义（P〈0．05）；肌电图受累神经部位〉t3的CIP患者抗GM1抗体水平高于受累神经部位≤2的CIP患者，差异亦有统计学意义（P〈0．05）。结论抗GM1 IgG抗体可以作为CIP诊断的血清学参考指标，其抗体水平与肌电图显示的神经损伤数目有关联，损伤程度越重抗GM1 IgG抗体水平越高。     

Sphingomyelin accumulation provides a favorable milieu for GM1 ganglioside-induced assembly of amyloid β-protein

The assembly of amyloid β-protein into fibrils is an initial event of Alzheimer's disease (AD). Previous studies suggest that ganglioside-bound amyloid β-protein (Aβ), GAβ, is an endogenous seed for amyloid in Alzheimer's disease (AD) brain and that GAβ is generated in the membrane microdomains, comprising cholesterol, sphingomyelin (SM) and GM1 ganglioside. In this study, we showed that the GAβ-dependent amyloidogenesis was accelerated on the surface of PC12 cells that had been pretreated with a sphingomyelinase inhibitor. Conversely, the enhanced GAβ-dependent amyloidogenesis under the endocytic dysfunction, which is one of the cell-pathological features of AD, was suppressed by pretreatment with a SM synthase inhibitor. These suggest that SM is one of the key molecules for GAβ generation and further imply that the interaction of Aβ with membrane lipids is critical in amyloid fibrillization in the brain.     

Targeting of G(D2)-positive tumor cells by human T lymphocytes engineered to express chimeric T-cell receptor genes.

Genetic engineering of human T lymphocytes to express tumor antigen-specific chimeric immune receptors is an attractive means for providing large numbers of effector cells for adoptive immunotherapy while bypassing major mechanisms of tumor escape from immune recognition. We have applied this strategy to the targeting of a G(D2)-positive tumor, neuroblastoma, which is the commonest extracranial solid tumor of childhood. Chimeric immune receptors were generated by joining an extracellular antigen-binding domain derived from either of the 2 ganglioside G(D2)-specific antibodies sc7A4 and sc14.G2a to a cytoplasmic signaling domain. The variable domains of hybridoma antibody 14.G2a were cloned and selected using a phage display approach. Upon coincubation with G(D2)-expressing tumor cell targets, human T lymphocytes transduced with recombinant retroviruses encoding chimeric receptors based on sc14.G2a, but not sc7A4, secreted significant levels of cytokines in a pattern comparable to the cytokine response obtained by engagement of the CD3 receptor. T cells transduced with the sc14.G2a-based chimeric T-cell receptors also displayed specific lysis of G(D2)-positive neuroblastoma cells, which was blocked in the presence of monoclonal antibody 14.G2a. In the absence of nonspecific stimulation of transduced cells, their functionality declined over time and antigenic stimulation of the chimeric receptor alone did not induce commitment to proliferation. These results support the feasibility of redirecting human T lymphocytes to a tumor-associated ganglioside epitope but emphasize that successful chimeric receptor-mediated adoptive immunotherapy will require additional strategies that overcome functional inactivation of gene-modified primary T lymphocytes.     

神经节苷脂治疗吉兰-巴雷综合征疗效及肌电图变化的临床评价

目的:评价神经节苷脂在促进吉兰-巴雷患者恢复方面上的临床疗效及肌电图变化。方法:将符合入选标准的36例患者随机分为2组,分别采用神经节苷脂和普通B族维生素治疗。治疗前后对两组患者定期进行MRC、Hughes评分以及肌电图检查,测定运动神经传导速度及F波平均潜伏期。结果:两组MRC、Hughes评分与治疗前相比均有显著或极显著差异(P<0.05-0.01),半年后两组MRC、Hughes评分比较有显著差异(P<0.05),半年后两组运动神经传导速度、F波潜伏期比较有显著差异(P<0.05),两组无明显不良反应。结论:神经节苷脂是促进吉兰-巴雷患者恢复的有效药物,无论是临床评分还是肌电图改善均优于普通B族维生素。