双上肢去势联合椎间盘刺破诱导建立大鼠椎间盘退变模型  

Establishing a rat model of intervertebral disc degeneration by castration of both upper limbs combined with intervertebral disc puncture

作　　者：孙孝先 白雪 刘孟敏 郭杨[1] 林顺 吴雯轩 马勇[1,2] 刘锦涛[3] Sun Xiaoxian;Bai Xue;Liu Mengmin;Guo Yang;Lin Shun;Wu Wenxuan;Ma Yong;Liu Jintao(Laboratory of New Techniques of Restoration&Reconstruction of Orthopedics and Traumatology,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu Province,China;School of Chinese Medicine/School of Integrated Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu Province,China;Suzhou Hospital of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Suzhou 215008,Jiangsu Province,China)

机构地区：[1]南京中医药大学骨伤修复与重建新技术实验室,江苏省南京市210023 [2]南京中医药大学中医学院·中西医结合学院,江苏省南京市210023 [3]南京中医药大学附属苏州市中医医院,江苏省苏州市215008

出　　处：《中国组织工程研究》2023年第35期5616-5621,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(82074467),项目负责人:刘锦涛;2022年大学生创新创业训练计划省级重点项目(202210315108Z),项目负责人:吴雯轩。

摘　　要：背景:寻求一种相对合适的椎间盘退变动物模型,对深入研究椎间盘退变的发病机制有着重要意义。目的:探索一种新的椎间盘退变动物模型,希望为椎间盘退变疾病的基础研究提供新的造模方式选择。方法:将20只雄性SD大鼠随机分为假手术组与模型组。采用双上肢去势联合椎间盘刺破的方式制备椎间盘退变模型,于造模当天及造模后4周、8周进行MRI检查,观察造模前后椎间盘形态变化并评估Pfirrmann分级;造模4,8周后取出大鼠L4-L5及L5-L6椎间盘,运用苏木精-伊红染色、Masson染色法观察椎间盘病理学变化及免疫组化观察Ⅱ型胶原蛋白表达量。结果与结论:①模型组在造模当天仅呈现轻微退变;②在造模第4周时与假手术组相比,模型组MRI上T2加权信号降低,在Pfirrmann分级上可见明显差异(P<0.05),在造模第8周时与假手术组相比,模型组MRI上T2加权信号明显降低,与纤维环边界模糊不清,且髓核向后纵韧带膨出,在Pfirrmann分级上可见显著差异(P<0.01);③病理染色结果显示,在造模第4周时,与假手术组相比,模型组髓核细胞减少,纤维环排列紊乱,Ⅱ型胶原表达量下降;在造模第8周时,与假手术组相比,模型组髓核细胞皱缩、基质凝结,纤维环破裂,结构遭到破坏且与髓核界限模糊,髓核部分Ⅱ型胶原表达量下降更为明显;④此方法可成功建立大鼠椎间盘退变模型,且动物模型退变明显,为研究椎间盘退变模型提供一种新思路。BACKGROUND:Seeking a relatively suitable animal model of intervertebral disc degeneration is of great significance for an in-depth study of the pathogenesis of intervertebral disc degeneration.OBJECTIVE:To seek a relatively suitable animal model of intervertebral disc degeneration,providing a new modeling option for basic research into degenerative disc disease.METHODS:Twenty male Sprague-Dawley rats were randomly divided into sham operation group and model group.The intervertebral disc degeneration model was prepared by castration of both upper limbs combined with intervertebral disc puncturing.MRI examination was performed on the day of modeling and at 4 and 8 weeks after modeling to observe the morphological changes of the intervertebral discs and evaluate the Pfirrmann grade before and after modeling.The L4-L5 and L5-L6 intervertebral discs were removed from the rats at 4 and 8 weeks after modeling.The pathological changes of the intervertebral discs were observed by hematoxylin-eosin and Masson staining and the expression of type II collagen was observed by immunohistochemistry.RESULTS AND CONCLUSION:The model group showed only slight degeneration on the day of modeling.Compared with the sham operation group,the T2-weighted signal on MRI was decreased in the model group,and there was a significant difference in the Pfirrmann grading at 4 weeks after modeling(P<0.05).Compared with the sham operation group,the T2-weighted signal on MRI was significantly decreased and the boundary with the annulus fibrosa was blurred in the model group at 8 weeks after modeling.The nucleus pulposus in the model group was expanded in the posterior longitudinal ligament and there was a significant difference in the Pfirrmann grading between the two groups(P<0.01).The results of pathological staining showed that compared with the sham operation group,the model group had a reduction in the number of nucleus pulposus cells,disordered annulus fibrosus,and decreased expression of type II collagen at the 4th week after modeling.While a

关 键 词：椎间盘退变 动物模型 大鼠 双上肢去势 纤维环穿刺 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R-332