环指蛋白RING1在大鼠脊髓损伤后的表达变化  

Spatiotemporal patterns of RING1 expression after rat spinal cord injury

作　　者：刘汉章[1] 计巍[2] 刘春[3] 刘晓娟[4] 张冬梅[4] 朱顺星[3] LIU Hanzhang JI Wei LIU Chun LIU Xiaojuan ZHANG Dongmei ZHU Shunxing(Morphology Laboratory Key Laboratory of Neuroregeneration, Nantong University Experimental Animal Center Key Laboratory for Inflammation and Molecular Drug Target, Medical College of Nantong University, Nantong 226001)

机构地区：[1]南通大学医学院形态学实验室,南通226001 [2]南通大学神经再生重点实验室,南通226001 [3]南通大学医学院实验动物中心,南通226001 [4]南通大学炎症与分子药靶重点实验室,南通226001

出　　处：《南通大学学报（医学版）》2016年第6期518-521,共4页Journal of Nantong University(Medical sciences)

基　　金：国家自然科学基金资助项目(31500647);南通大学自然科学研究基金资助项目(13040510)

摘　　要：目的:研究环指蛋白RING1在脊髓损伤(spinal cord injury,SCI)中的变化和作用。方法:用Basso Beattie Bresnahan(BBB)运动功能评分来判断急性SCI后的运动功能恢复情况;免疫蛋白印迹试验研究RING1在SCI后的蛋白表达变化;逆转录-聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)检测RING1在SCI后的m RNA表达情况;免疫荧光染色检测SCI后的RING1和神经细胞特异性标志物:神经元(Neu N),星形胶质细胞[胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)],小胶质细胞[分化群分子11b(cluster of differentiation molecule 11b,CD11b)]共定位细胞的变化。结果:所有动物在SCI后出现松弛的后肢运动,随着时间的推移运动功能慢慢恢复;损伤后1 d RING1的m RNA水平有显著增高,然后逐渐下降到正常水平;在损伤后3 d RING1的蛋白量表达显著升高,随后表达下降;损伤后3 d RING1在星型胶质细胞中表达有明显增加,而在神经元、小胶质细胞中的表达并无明显的变化。结论:RING1在SCI后的星形胶质细胞中表达明显增加,表示其可能在SCI中扮演重要角色。Objective： To explore the changes and effects of RING1 after spinal cord injury（SCI）. Metholds： The spontaneous recovery of locomotor function after SCI was examined using the Basso, Beattie and Bresnahan（BBB） rating scale. RING1 protein level was detected by Western Blot and mRNA level was investigated by reverse transcription-polymerase chain reaction （RT-PCR） after SCI. Immunofluorescence staining was performed to observe the changes of double-labeled cells for RING1 and the nerve cell phenotypic markers： NeuN for neurons, glial fibrillary acidic protein （GFAP） for astrocytes, cluster of differentiation molecule l lb（CDllb） for microglias. Results： All animals slacked hind limb locomotion after contusion. Some recoveries of spontaneous function showed up with time after SCI. RING1 mRNA level was strongly up-regulated after contusion and peaked at day 1 following SCI, and reduced gradually to the normal level. Furthermore, RING1 protein level was peaked at day 3 and then gradually reduced. The expression of RING1 increased more significantly in astrocytes, while less significantly in neurons and microglia at day 3 after injury. Conclusion： The expression of RING1 after spinal cord injury was significantly increased in astrocytes, which indicates that it may play an important role in spinal cord injury.

关 键 词：脊髓损伤 RING1 星形胶质细胞 免疫蛋白印迹试验 免疫荧光 大鼠 

分 类 号：R683.2[医药卫生—骨科学]