一氧化碳损伤后少突胶质细胞及其相关神经营养因子Lingo-1的变化  

The changes of oligodendrocytes and associated neurophic factor of Lingo - 1 after carbonmonoxide injury

作　　者：马英[1] 王天双 肖刚[2] 李林[1] 张德新[1] 张俭[1] 陆莹[1] 

机构地区：[1]哈尔滨医科大学附属第二医院急诊科,黑龙江哈尔滨150080 [2]中国人民解放军第二一一医院,黑龙江哈尔滨150080

出　　处：《中国急救医学》2017年第7期642-645,I0003,共5页Chinese Journal of Critical Care Medicine

基　　金：黑龙江省自然科学基金资助项目（H201422）

摘　　要：目的探讨一氧化碳（carbonmonoxide，CO）损伤后少突胶质细胞及其相关神经营养因子富含亮氨酸重复序列和免疫球蛋白结构域的Nogo受体作用蛋白-1（1eucinerichrepeatandIgdomaincontaining，nogoreceptorinteractingprotein-1，Lingo-1）的变化情况。方法取新生（36h内）SD大鼠脑皮质进行体外培养，采用细胞贴壁时间差分离纯化法和条件限定培养基纯化少突胶质细胞。倒置显微镜结合免疫荧光染色鉴定细胞种类，观察少突胶质细胞的形态变化。培养细胞随机分为对照组与CO损伤组，对照组细胞正常培养；CO损伤组细胞置于含体积分数1％CO的密闭三气培养箱内培养。CO处理后0、12、24、48、72h各时间点逆转录聚合酶链式反应检测两组Lingo-1mRNA的表达量。结果①获取高纯度的少突胶质细胞，MBP及Lingo-1免疫荧光染色阳性。②CO损伤后，少突胶质细胞存活率明显下降，并且随着时间延长存活细胞减少，伴凋亡细胞增多，至72h多数细胞破碎。③与对照组相比较，Lingo-1mRNA在CO损伤组中的表达量高于对照组（P〈0．05），至CO处理后48h达峰值（P〈0．05）。结论CO损伤后，体外培养的少突胶质细胞存活率明显下降，且Lingo-1的表达上调，提示CO中毒过程中Lingo-1与大脑脱髓鞘病变相关。Objective To investigate the changes of oligodendrocytes and associated neurophic factors after carbon monoxide injury. Related neurophic factor investigated in present study was Lingo - 1 （ leucine - rich repeat - and Ig domain - containing nogo receptor - interacting protein - 1 ）. Methods Took new （36 hours） SD rats＇ cortex in vitro and purified oligodendrocytes by cell adherent time purification method and the condition of limited medium purification. Inverted microscope combined with immunofluorescence staining appraisal eel｜ types, observing the morphdtogy changes of oligodendrocytes. Cells were randomly divided into two groups, control group and CO injured group. In the control group ceils were cultured normally. In CO injured group ceils were cultured in the three gas airtight incubated with 0.1% CO. The mRNA expressions of Lingo - 1 were detected by reverse transcription - polymerase chain reaction at 0 h, 12 h, 24 h, 48 h and 72 h after given with CO. Results Theimmunofluorescence staining of Lingo - 1 and MBP were positive. After the treatment with CO the survival rate of oligodendrocytes decreased significantly and ceils apoptosis increased with time. It was until 72 h that most cells were broken. Compared with control group, the mRNA expressions of Lingo - 1 was higher in the CO injured group （ P 〈 0.05 ） with the highest quantity at 48 h （ P 〈 0.05 ）. Conclusion The survival rate of oligodendrocytes after the injury of carbon monoxide in vitro decreased obviously relating with the expressions of Lingo - 1, which meant that Lingo - 1 may be related to the damages of oligodendrocytes and further brain demyelinating changes.

关 键 词：一氧化碳中毒 迟发性脑病 少突胶质细胞 LINGO-1 

分 类 号：R774.6[医药卫生—眼科]