小鼠髓核特异性标志物的鉴定和表达分析  

Identification and expression analysis of mouse nucleus pulposus specific markers

作　　者：张磊 修春美 倪莉[2] 陈建权 Zhang Lei;Xiu Chunmei;Ni Li;Chen Jianquan(Institute of Orthopedics,Medical College of Soochow University,Suzhou 215008,Jiangsu Province,China;Department of Orthopedics,The First Affiliated Hospital of Soochow University,Suzhou 215006,Jiangsu Province,China)

机构地区：[1]苏州大学医学部骨科研究所,江苏省苏州市215008 [2]苏州大学附属第一医院骨科,江苏省苏州市215006

出　　处：《中国组织工程研究》2022年第11期1669-1674,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81902248),项目负责人:倪莉。

摘　　要：背景:椎间盘退变是下腰痛的主要诱因,而髓核细胞在椎间盘退变中起着至关重要的作用。小鼠模型是研究椎间盘稳态和退变的重要动物模型,然而对小鼠髓核特异性标志物目前还缺乏较为系统的分析。目的:验证细胞角蛋白19、葡萄糖转运蛋白1和磷脂酰肌醇蛋白聚糖3这3种蛋白在不同时期小鼠椎间盘内的表达情况及其细胞特异性;探讨上述蛋白作为小鼠髓核特异性标志物的可行性。方法:10只6周龄C57BL/6雄性小鼠随机分为2月龄组及12月龄组(n=5),分别饲养至相应月龄时收取椎间盘样本进行后续分析。采用苏木精-伊红染色和番红固绿染色评估椎间盘组织学完整形态;利用免疫荧光或免疫组化染色检测细胞角蛋白19、葡萄糖转运蛋白1和磷脂酰肌醇蛋白聚糖3这3种蛋白在两组小鼠椎间盘冠状位切片上的表达情况,并通过统计学方法分析各蛋白在不同年龄小鼠椎间盘中的表达变化。结果与结论:①在2月龄小鼠椎间盘内,上述3种标志物均可明显区分髓核与其他组织;②在12月龄老年小鼠椎间盘内,只有细胞角蛋白19仍旧特异性表达于髓核组织,而葡萄糖转运蛋白1和磷脂酰肌醇蛋白聚糖3除在髓核细胞表达外,在软骨终板的钙化处亦有出现;③提示细胞角蛋白19、葡萄糖转运蛋白1和磷脂酰肌醇蛋白聚糖3均在小鼠髓核细胞表达,其中细胞角蛋白19可作为年轻及老龄小鼠的髓核特异性标志物。BACKGROUND:Intervertebral disc degeneration is the main cause of low back pain,and nucleus pulposus cells play a vital role in intervertebral disc degeneration.A mouse model is an important animal model for studying the homeostasis and degeneration of the intervertebral disc.However,there is still a lack of systematic analysis of mouse nucleus pulposus specific markers.OBJECTIVE:To verify the expression and cell specificity of cytokeratin 19,glucose transporter 1,and glypican 3 in the intervertebral discs of mice at different stages and to explore the feasibility of the above proteins used as specific markers of the mouse nucleus pulposus.METHODS:Ten 6-week-old C57 BL/6 male mice were randomly divided into a 2-month-old group and a 12-month-old group(n=5),and intervertebral disc samples were collected for subsequent analysis when the mice were raised to the corresponding months.Hematoxylin-eosin staining and Safranin O&Fast Green staining were used to evaluate the histological integrity of the intervertebral discs.Immunofluorescence or immunohistochemical staining was used to detect the protein expression of cytokeratin 19,glucose transporter 1,and glypican 3 on the coronal slices of the intervertebral discs,and protein expression changes in mice of different ages were statistically analyzed.RESULTS AND CONCLUSION:Cytokeratin 19,glucose transporter 1,and glypican 3 were specifically expressed in the nucleus pulposus tissues of 2-month-old mice.Only cytokeratin 19 was specially expressed in the nucleus pulposus tissues of 12-month-old mice,whereas glucose transporter 1 and glypican 3 appeared in the nucleus pulposus cells as well as in the endplate.Therefore,cytokeratin 19,glucose transporter 1,and glypican 3 can all be expressed in the nucleus pulposus tissues from 2-and 12-month-old mice,but only cytokeratin 19 can serve as the nucleus pulposus-specific marker for both young and aged mice.

关 键 词：髓核 细胞角蛋白19 葡萄糖转运蛋白1 磷脂酰肌醇蛋白聚糖3 椎间盘退变 小鼠 髓核标志物 

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