摘要
目的通过终板下注射无水乙醇阻碍椎体-终板营养,建立一种新型兔腰椎椎间盘退行性变模型,并观察终板退行性变过程中内皮素1(ET-1)的表达情况。方法健康4月龄新西兰兔32只,随机分成4组,每组8只,选取L5,6椎体(对应L4/L5及L5/L6椎间盘)注射300μL无水乙醇,选取L4椎体(对应L3/L4椎间盘)注射磷酸盐缓冲液(PBS)作为实验对照,L7椎体(对应L6/L7椎间盘)未注入任何物质作为正常对照。其中1组造模后1个月提取软骨终板细胞,行免疫细胞化学染色检测ET-1表达;余3组分别于造模后1、3和5个月进行椎间盘X线和MRI检查,取椎间盘组织行HE染色观察形态学改变,免疫组织化学染色观察ET-1表达。结果注射无水乙醇后,随着时间进展,X线片显示椎间隙高度显著下降、椎间隙变窄、边缘骨赘增生,MRI T2WI显示椎间盘低信号;苏木精-伊红染色(HE)显示终板的生长板厚度变薄,终板结构破损,同时软骨终板细胞退化、直至消失,髓核中细胞发生转化(由空泡细胞转变为软骨样细胞,进而形成纤维软骨样细胞)造成髓核纤维化,纤维环结构排列紊乱、纤维化程度逐步加重;免疫组织化学染色显示,发生退行性变的终板组织内有ET-1表达,但随着退行性变加剧,ET-1表达强度下降;提取的退行性变软骨终板细胞(造模后1个月)也显示细胞质内ET-1强表达。结论通过注射无水乙醇阻碍椎体-终板营养途径可成功建立兔椎间盘退行性变模型,终板退行性变过程中伴随ET-1的表达。
Objective To establish a new model of lumbar disc degeneration by injecting absolute ethanol into the vertebrae body adjacent to the endplate to affect the nutritional supply of the intervertebral disc,and to evaluate the expression of endothelin-1(ET-1) in the process of endplate degeneration.Methods Thirty-two New Zealand white rabbits aged 16 weeks were included in the study,which were divided into 4 groups at random(n=8).300 μL absolute ethanol was injected into the L5,6 vertebral body close to sub-endplate,respectively,so the corresponding intervertebral discs were L4/L5 and L5/L6 discs.Similarly,L4 vertebral body was injected with 300 μL of phosphate-buffered saline(PBS),and corresponding to L3/L4 disc as experimental control.The L7 vertebra body(corresponding to L6/L7 disc) was not injected with any substance as the normal control.One group was used to extract cartilaginous endplate cells(CECs) at 1 month after surgery.Intervertebral disc degeneration was analyzed by X-ray,MRI and HE staining,and ET-1 expression was detected by immunohistochemical staining after the operation at the time point of 1,3 and 5 months in the other 3 groups.Results With the progress of time after absolute ethanol injection,roentgenograph showed a significant decrease in height of intervertebral disc accompanied by intervertebral space narrow,and osteophyte formation close to vertebral body edge in the absolute ethanol injected group.The MRI T2WI showed low signals and gradually worsening disc degeneration.Hematoxylin-Eosin(HE) staining showed that the extent of endplate degeneration increased gradually over time including decreased thickness of growth plate,and structural damage of the endplate.At the same time,CECs degenerated and eventually disappeared.In the nucleus pulposus,cells underwent transformation mainly from vacuolar cells to chondroid cells,and eventually formed fibrocartilage-like cells along with nucleus pulposus fibrosis.With the progression of degeneration,the annulus fibrosus demonstrated disordered structura
作者
袁维
周小小
蔡攀
陈诚
郭胜洋
吴佳俊
钱明
YUAN Wei;ZHOU Xiao-xiao;CAI Pan;CHEN Cheng;GUO Sheng-yang;WUJia-jun;QIAN Ming(Department of Orthopaedics,Shanghai Fourth People's Hospital,Tongji University School of Medicine,Shanghai 200081,China;Department of Orthopaedics,Zhoupu Hospital,Shanghai University of Medicine&Health Sciences,Shanghai 201318,China;Department of Orthopaedics,Changzheng Hospital,Navy Medical University,Shanghai 200003,China)
出处
《脊柱外科杂志》
2020年第2期118-124,共7页
Journal of Spinal Surgery
基金
上海市自然科学基金项目(14ZR1442300)
上海市卫生和计划生育委员会科研课题面上项目(201440387)
上海市浦东新区卫生系统学科带头人培养计划(PWRd201502)
上海健康医学院师资人才百人库培养计划(ZPBRK-18-09)。
关键词
腰椎
椎间盘退行性变
内皮素-1
模型
动物
兔
Lumbar vertebrae
Intervertebral disc degeneration
Endothelin-1
Models,animal
Rabbits
