CAJ | 학술논문

目的观察嗅球成鞘细胞(olfactory ensheathing cells,OECs)在损伤脊髓内的成髓鞘作用及其对髓鞘形成的影响. 方法用NYU-Ⅱ型脊髓撞击器(强度为10 g×25 mm)损伤大鼠T10脊髓后1周,将绿色荧光蛋白(green fluorescence protein,GFP)-OECs植入脊髓损伤部位及其首尾两侧,设为OECs移植组;以同量改良Eagle培养基(Dulbecco's modification of Eagle's medium,DMEM)注射作为对照组.移植后6周,取出脊髓做冰冻切片,行髓鞘碱性蛋白(myelin basic protein,MBP)、0蛋白(protein zero,P0)和S100蛋白(S100 protein,S100)免疫细胞化学染色,定性和半定量观察免疫染色情况;并行塑料包埋后半薄和超薄切片,先于光镜下进行定性和半定量观察,再于电镜下观察髓鞘的超微结构. 结果 OECs移植组S100、MBP、P0所显示的有髓神经纤维由正常组织向损伤区延伸,并可贯穿损伤区,所显示的纤维分别占损伤区面积的12.3％、11.6％和9.3％,且3种蛋白显示的神经纤维数量差异无统计学意义,但均显著高于对照组(2.89％)(P＜0.01).OECs移植组半薄切片上移植组损伤区内有髓神经纤维的数量为(354.67±59.00),较对照组的(167.33 ±42.16)明显增多(P＜0.01),OECs移植组形成的髓鞘较对照组直径小、髓鞘薄. 结论 OECs可以促进再生神经纤维髓鞘形成,部分OECs自身形成髓鞘,但髓鞘结构较薄.
목적 관찰후구성초세포(olfactory ensheathing cells,OECs)재손상척수내적성수초작용급기대수초형성적영향. 방법 용NYU-Ⅱ형척수당격기(강도위10 g×25 mm)손상대서T10척수후1주,장록색형광단백(green fluorescence protein,GFP)-OECs식입척수손상부위급기수미량측,설위OECs이식조;이동량개량Eagle배양기(Dulbecco's modification of Eagle's medium,DMEM)주사작위대조조.이식후6주,취출척수주빙동절편,행수초감성단백(myelin basic protein,MBP)、0단백(protein zero,P0)화S100단백(S100 protein,S100)면역세포화학염색,정성화반정량관찰면역염색정황;병행소료포매후반박화초박절편,선우광경하진행정성화반정량관찰,재우전경하관찰수초적초미결구. 결과 OECs이식조S100、MBP、P0소현시적유수신경섬유유정상조직향손상구연신,병가관천손상구,소현시적섬유분별점손상구면적적12.3％、11.6％화9.3％,차3충단백현시적신경섬유수량차이무통계학의의,단균현저고우대조조(2.89％)(P＜0.01).OECs이식조반박절편상이식조손상구내유수신경섬유적수량위(354.67±59.00),교대조조적(167.33 ±42.16)명현증다(P＜0.01),OECs이식조형성적수초교대조조직경소、수초박. 결론 OECs가이촉진재생신경섬유수초형성,부분OECs자신형성수초,단수초결구교박.
Objective To detect the myelinating role of olfactory ensheathing cells (OECs in the contused spinal cord and their impact on remyelination.Methods The rats were subjected to spinal cord injury at T10(10 g ×25 mm) using a NYU-Ⅱ impactor.One week later,the rats were transplanted with green fluorescence protein (GFP)-OECs (OECs group) or an equal volume of Dulbecco' s modification of Eagle's medium (DMEM) (control group) at epicenter of the injury as well as its rostral and caudal sites.Six weeks after transplantation,the spinal cords were removed for frozen section.Myelin basic protein (MBP),protein zero (P0),and S100 protein (S100) were determined with qualitative and semi-quantitative immunocytochemical assay.Moreover,plastic embedded semithin and ultrathin sections were prepared for qualitative and semi-quantitative examination under light microscopy and electroscopic study of myelin sheath ultrastructure.Results In OECs group,the nerve fibers labeled with S100,MBP,and PO were extended from the normal tissues to the injured region and even grew through the region with space consuming of 12.3％,11.6％,and 9.3％ respectively.Moreover,there were no statistical differences regarding the number of fibers labeled by the three proteins,but all were significantly larger than that in control group (2.89％,P ＜ 0.01).Number of myelinated nerve fibers in injured regions on hemithin sections was increased significantly to 354.67 ± 59.00 in OECs group,with significant difference compared with 167.33 ± 42.16 in control group (P ＜ 0.01).The regenerated myelin sheaths in OECs group were smaller and thicker than those in control group.Conclusions OECs can accelerate regeneration of myelinated nerve fibers.Additionally,some OECs form myelin sheaths themselves,but the sheath structures are relatively thinner.