CAJ | 학술논문

目的探讨Sirt1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Sirt1、Ⅱ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Sirt1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,Sirt1基因表达显著下降.
목적 탐토Sirt1기인재인경추종판연골세포퇴변과정중적작용급의의.방법 선취2012년1월지2012년7월우환남의학원익기산의원척주외과주원접수경전로추체차전절제수술적경추퇴행성질병환자급외상성경추골절、탈위혹척수손상환자30례.분위경추병조(14례)화대조조(16례),건립인경추추체종판연골세포퇴변모형.채용매소화법분리급배양종판연골세포,현미경하관찰세포형태,이용HE염색급갑분알람염색대종판연골세포진행관찰급감정.운용실시역전록취합매련반응급면역인적법검측종판연골세포퇴변과정중Sirt1、Ⅱ형효원급단백다당기인적표체변화.결과 퇴변조여정상조종판연골세포상비교,세포출현사형변추세.Ⅱ형효원기인(P=0.0072)급단백다당기인(P =0.0063) mRNA적표체균상응감저.퇴변조Sirt1기인표체교정상조명현강저(P =0.034).결론 인경종판연골세포발생퇴변후,Sirt1기인표체현저하강.
Objective To observe the expression changes of Sirt1 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 Sirt1,collagen Ⅱ and aggrecan.Results Compared with the normal group,the cellular morphology of degenerative group showed spindle-shaped changes.The mRNA expression of Sirt1 (P =0.034) significantly decreased.Aggrecan (P =0.0063) and collagen Ⅱ (P =0.0072)decreased also markedly.Conclusion Sirt1 gene expression is significantly down-regulated in degenerative human cervical endplate chondrocytes.Regulating the expression of Sirt1 gene may block or delay the occurrence of human cervical endplate cartilage degeneration.