神经节苷脂对体外培养一氧化碳损伤少突胶质细胞Nogo-A的影响

背景:Nogo-A蛋白是表达于少突胶质细胞的神经元轴突生长抑制因子,推测其可能与一氧化碳中毒后迟发性脑病关系密切。单唾液酸四己糖神经节苷脂可改善一氧化碳中毒所致的神经系统损害,其作用是否与少突胶质细胞上Nogo-A蛋白有关目前尚无报道。目的:从少突胶质细胞Nogo-A的角度探讨神经节苷脂对神经细胞一氧化碳损伤后保护的机制。方法:体外培养大鼠视神经少突胶质细胞。实验分为3组:对照组、一氧化碳组、单唾液酸四己糖神经节苷脂组。后两组在含有体积分数1%一氧化碳的密室中培养,单唾液酸四己糖神经节苷脂组预先在培养液中加入5 mg/L单唾液酸四己糖神经节苷脂。采用RT-PCR及细胞免疫组织化学观察6,24,48 h少突胶质细胞Nogo-A mR NA及其蛋白质的表达。结果与结论:一氧化碳组6,24及48 h各间点Nogo-A mR NA积分吸光度比值、Nogo-A蛋白质表达的累计吸光度值均显著高于对照组(P<0.05);一氧化碳处理后24 h,单唾液酸四己糖神经节苷脂组Nogo-A mR NA积分吸光度比值、蛋白质表达的累计吸光度值均显著高于对照组(P<0.05),但低于一氧化碳组(P<0.05);Nogo-A mR NA水平与蛋白质表达具有线性相关性(r=0.95)。提示一氧化碳本身可使少突胶质细胞进入活跃的功能状态,提高Nogo-A的转录和翻译水平;Nogo-A蛋白质表达的增加为其mR NA水平水平提高所致;单唾液酸四己糖神经节苷脂对神经细胞急性一氧化碳损伤的保护作用与抑制少突胶质细胞Nogo-A表达有关。     

大鼠视神经少突胶质细胞培养方法的改良

目的改良既往新生大鼠视神经组织块培养法体外培养少突胶质细胞。方法新生2 d SD大鼠,无菌条件下取双侧视神经,置于预先涂有多聚赖胺酸的培养皿中（直径35 cm）,加入基础培养液约400 μL;培养第4天时,更换为含05%胎牛血清化学限定培养液约400 μL;约第10天时,更换为无胎牛血清化学限定培养液约600 μL;第11天行髓鞘碱性蛋白（myelin basic protein,MBP）细胞免疫化学鉴定,计算阳性细胞百分率。重复培养3次,方差分析方法的稳定性。结果视神经培养至第11天,每皿细胞数可达(6~8)×105个,90％以上为MBP阳性细胞;3次培养细胞的MBP免疫细胞化学染色阳性细胞百分率差异无统计学意义（P＞005）。结论本实验方法所得成熟少突胶质细胞纯度高,数量足够一般细胞生物学实验使用,且方法简便、稳定。     

血红素氧合酶系统对一氧化碳中毒后体外培养少突胶质细胞Nogo-A的影响

BACKGROUND: Cerebral white matter demyelination is outstanding in the images of delayed encephalopathy after acute carbon monoxide (CO) poisoning. Since Nogo-A and Nogo-receptor are expressed in onoligodendrocytes and neurons respectively, we infer that Nogo-A system is involved in brain injury after acute CO poisoning and related to delayed encephalopathy after acute CO poisoning. Endogenous CO is a gaseous messenger, which is the metabolic product of hemeoxygenase. There is no report about the CO effect on Nogo-A system till now.OBIECTIVE:To in vitro culture oligodendrocytes using endogenous CO, inhibit the activity of hemeoxygenase system using zinc protoporphyrin-IX (ZnPPIX) and observe the variation of Nogo-A in oligodendrocytes at mRNA and protein levels.METHODS:Rat oligodendrocytes cultured in vitro were divided into control, CO, ZnPPIX groups. Cells in the CO and ZnPPIX groups were treated with 1% CO directly, In the ZnPPIX group, 10 μmol/L ZnPPIX was added into the culture medium before CO treatment. The expressions of Nogo-A mRNA and protein at 6, 24, 48 hours after culture were compared. Differences in the peak levels of Nogo-A mRNA and protein between CO and ZnPPIX groups were detected using RT-PCR and immunohistochemistry respectively.RESULTS:The expression levels of Nogo-A mRNA and protein were significantly higher in the CO group than the control group and reached the peak at 24 hours of culture. Compared with the CO group, oligodendrocytes cultured with ZnPPIX showed higher expressions of Nogo-A mRNA and protein at 24 hours of culture. These findings suggest that except the influence of hypoxia occurring in CO poisoning, exogenous CO increases the expression of Nogo-A in cultured oligodendrocytes in vitro, and the heme oxygenase system can inhibit the expression of Nogo-A mRNA and protein.     

神经节苷脂对体外培养一氧化碳损伤少突胶质细胞Nogo-A的影响

背景：Nogo-A蛋白是表达于少突胶质细胞的神经元轴突生长抑制因子,推测其可能与一氧化碳中毒后迟发性脑病关系密切。单唾液酸四己糖神经节苷脂可改善一氧化碳中毒所致的神经系统损害,其作用是否与少突胶质细胞上Nogo-A蛋白有关目前尚无报道。 目的：从少突胶质细胞Nogo-A的角度探讨神经节苷脂对神经细胞一氧化碳损伤后保护的机制。 方法：体外培养大鼠视神经少突胶质细胞。实验分为3组：对照组、一氧化碳组、单唾液酸四己糖神经节苷脂组。后两组在含有体积分数1%一氧化碳的密室中培养,单唾液酸四己糖神经节苷脂组预先在培养液中加入5 mg/L单唾液酸四己糖神经节苷脂。采用RT-PCR及细胞免疫组织化学观察6,24,48 h少突胶质细胞Nogo-A mRNA及其蛋白质的表达。 结果与结论：一氧化碳组6,24及48 h各间点Nogo-A mRNA积分吸光度比值、Nogo-A蛋白质表达的累计吸光度值均显著高于对照组(P < 0.05);一氧化碳处理后24 h,单唾液酸四己糖神经节苷脂组Nogo-A mRNA积分吸光度比值、蛋白质表达的累计吸光度值均显著高于对照组(P < 0.05),但低于一氧化碳组(P < 0.05);Nogo-A mRNA水平与蛋白质表达具有线性相关性(r = 0.95)。提示一氧化碳本身可使少突胶质细胞进入活跃的功能状态,提高Nogo-A的转录和翻译水平;Nogo-A蛋白质表达的增加为其mRNA水平水平提高所致;单唾液酸四己糖神经节苷脂对神经细胞急性一氧化碳损伤的保护作用与抑制少突胶质细胞Nogo-A表达有关。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：