人颈椎椎体终板软骨细胞退变中的Sirtl基因表达及意义  

Significance and expression of Sirtl gene in human cervical endplate chondrocyte in vitro degeneration

作　　者：胡斌[1] 徐宏光[1] 宋俊兴[1] 章平治[1] 张小海 程加峰 

机构地区：[1]皖南医学院弋矶山医院脊柱外科,安徽省芜湖市241001 [2]芜湖市第二人民医院骨三科 [3]芜湖市第一人民医院骨科

出　　处：《中华医学杂志》2012年第47期3341-3344,共4页National Medical Journal of China

基　　金：国家自然科学基金（30973025,81272048）;安徽省教育厅自然科学基金（KJ2010A320）;卫生部公益性行业专项基金（201002018）

摘　　要：目的探讨Sirtl基因在人颈椎终板软骨细胞退变过程中的作用及意义。方法选取2012年1月至2012年7月于皖南医学院弋矶山医院脊柱外科住院接受颈前路椎体次全切除手术的颈椎退行性疾病患者及外伤性颈椎骨折、脱位或脊髓损伤患者30例。分为颈椎病组（14例）和对照组（16例），建立人颈椎椎体终板软骨细胞退变模型。采用酶消化法分离及培养终板软骨细胞，显微镜下观察细胞形态，利用HE染色及甲苯胺蓝染色对终板软骨细胞进行观察及鉴定。运用实时逆转录聚合酶链反应及免疫印迹法检测终板软骨细胞退变过程中Sirtl、11型胶原及蛋白多糖基因的表达变化。结果退变组与正常组终板软骨细胞相比较，细胞出现梭形变趋势。Ⅱ型胶原基因（P=0．0072）及蛋白多糖基因（P=0．0063）mRNA的表达均相应减低。退变组Sirtl基因表达较正常组明显降低（P=0．034）。结论人颈终板软骨细胞发生退变后，Sirtl基因表达显著下降。Objective To observe the expression changes of Sirtl gene and examine the role and significance of degenerative process in human cervical endplate chondrocytes through a degeneration model of human cervical vertebral endplate chondrocyte. Methods Cartilage endplates of 30 patients were divided into control group （ n = 16） with cervical vertebral fracture or dislocation and cervical spondylosis group （n = 14） with cervical spondylotic myelopathy. Endplate chondrocytes were isolated by enzyme digestion and cultured in vitro for 10 days. The differences of endplate chondrocytes from normal and degenerative cartilage endplates were observed by inverted phase-contrast microscope, hematoxylin and eosin staining and toluidine blue staining. Real-time reverse transcription-polymerase chain reaction （RT-PCR） and Western blot were used to detect the mRNA expressions of Sirtl, collagen I1 and aggrecan. Results Compared with the normal group, the cellular morphology of degenerative group showed spindle-shaped changes. The mRNA expression of Sirtl （ P = 0. 034 ） significantly decreased. Aggrecan （ P = 0. 0063 ） and collagen 1] （ P = 0. 0072 ） decreased also markedly. Conclusion Sirtl gene expression is significantly down-regulated in degenerative human cervical endplate chondrocytes. Regulating the expression of Sirtl gene may block or delay the occurrence of human cervical endplate cartilage degeneration.

关 键 词：板椎间盘 颈椎 软骨细胞 基因表达 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]