CAJ | 학술논문

目的观察RE-1元件辅助抑制因子(CoREST)在骨关节炎(OA)软骨细胞中的差异表达并探讨CoREST对OA软骨细胞分化的调控作用.方法取材接受膝关节表面置换的OA患者和正常对照组软骨并提取软骨细胞总蛋白,以Western blot对比CoREST在OA(n=8)和正常软骨(n=8)细胞中表达水平的差异.培养正常原代软骨细胞,采用针对CoREST设计的敲除siRNACoREST的表达至OA相当水平.应用Realtime聚合酶链反应(PCR)技术监测敲除CoREST前后软骨细胞(n=6)表型基因(Ⅰ型胶原、Ⅱ型胶原、aggrecan和X型胶原)表达水平的变化.结果 CoREST表达水平较正常软骨细胞下降69.5%(P＜0.01).将正常软骨细胞中的CoREST表达knock-down64.8%后,软骨细胞终末分化表型基因X型胶原的表达水平升高40.0%(P＜0.05),而正常分化表型Ⅱ型胶原和aggrecan的基因表达水平分别降低71.4%(P＜0.01)和57.6%(P＜0.01).结论 CoREST在OA软骨细胞中表达下降,诱发软骨细胞表型基因表达变化,提示CoREST参与了OA软骨细胞分化的调控.
목적 관찰RE-1원건보조억제인자(CoREST)재골관절염(OA)연골세포중적차이표체병탐토CoREST대OA연골세포분화적조공작용.방법 취재접수슬관절표면치환적OA환자화정상대조조연골병제취연골세포총단백,이Western blot대비CoREST재OA(n=8)화정상연골(n=8)세포중표체수평적차이.배양정상원대연골세포,채용침대CoREST설계적고제siRNACoREST적표체지OA상당수평.응용Realtime취합매련반응(PCR)기술감측고제CoREST전후연골세포(n=6)표형기인(Ⅰ형효원、Ⅱ형효원、aggrecan화X형효원)표체수평적변화.결과 CoREST표체수평교정상연골세포하강69.5%(P＜0.01).장정상연골세포중적CoREST표체knock-down64.8%후,연골세포종말분화표형기인X형효원적표체수평승고40.0%(P＜0.05),이정상분화표형Ⅱ형효원화aggrecan적기인표체수평분별강저71.4%(P＜0.01)화57.6%(P＜0.01).결론 CoREST재OA연골세포중표체하강,유발연골세포표형기인표체변화,제시CoREST삼여료OA연골세포분화적조공.
Objective To compare the differential expression of RE-1 silencing transcription factor (REST) compressor (CoREST) in normal and osteoarthritic chondrocytes and investigate the role of CoREST in regulating osteoarthritic chondrocyte differentiation. Methods Osteoarthritic knee cartilage samples were obtained from patients with osteoarthritis undergoing total knee replacement and normal controls.Chonrocytes were isolated and total protein was extracted for Western blotting analysis to compare the differential expression of CoREST between normal controls ( n = 8 ) and osteoarthritic ( n = 8 ) chondrocytes. Primarily cultured normal chondrocytes ( n = 6 ) were treated with specific siRNA to mimic CoREST repression in osteoarthritis. Realtime polymerase chain reaction (PCR) was used to compare the differential expression of chondrocyte phynotypic genes ( collagen Ⅰ, collagen Ⅱ, collagen X and aggrecan) before and after CoREST knock-down. Results CoREST expression level was down-regulated by 69. 5% (P ＜ 0. 01 ) in osteoarthritic chondrocytes in contrast to that of normal controls. In response to CoREST knock-down by 64. 8% (P ＜0. 01 ) in the primarily cultured chondrocytes, the gene expression level of the chondrocyte terminal differentiation marker gene, collagen X was up-regulated by 40. 0% ( P ＜ O. 05 ), whereas the differentiation phenotypic genes, collagen Ⅱ and aggrecan were down-regulated by 71.4% (P ＜0. 01 ) and 57.6% (P ＜0. 01 ), respectively. Conclusion CoREST down-regulation triggered the expression regulation of phenotypic genes towards terminal differentiation, which suggested CoREST is involved in the differentiation status modulation of osteoarthritic chodnrocytes.