CAJ | 학술논문

背景:软骨细胞具有修复受损软骨组织和维持软骨完整性的重要作用.线粒体功能障碍与细胞凋亡、老化和骨关节炎的病理过程密切相关.目的:通过线粒体 PCR 芯片技术研究骨性关节炎软骨细胞中线粒体基因的差异表达.方法:收集骨性关节炎患者及正常成人车祸后截肢者的关节软骨细胞,经过细胞的提取和培养、RNA 提取和质量检测、mRNA 提取和 cDNA 合成等处理后进行实时定量 PCR 检测.结果与结论:所检测到的84个与线粒体相关的待检测基因中有18个线粒体基因在骨性关节炎中发生了显著改变.以骨性关节炎患者软骨细胞线粒体基因相对正常成人软骨细胞线粒体基因的表达倍数表示基因表达的改变情况,其中倍数改变和倍数调节均大于2的上调基因有 BBC3,BCL2,SLC25A37等15个,倍数改变小于0.5和倍数调节小于-2的下调基因有 CPT1B,SLC25A16,SLC25A24共3个.18个线粒体差异基因功能分类如下:膜极化和电位:BCL2, BCL2L1, TP53, UCP1, UCP3;线粒体转运功能:BCL2, BCL2L1, CPT1B, FXC1(TIMM10B), MFN2, STARD3, TP53, UCP1, UCP3;小分子转运功能:SLC25A16, SLC25A2, SLC25A24, SLC25A31, SLC25A37;靶蛋白:FXC1(TIMM10B),MFN2;线粒体蛋白转入:COX18, FXC1(TIMM10B);内膜转运:FXC1(TIMM10B), TIMM17B;线粒体裂变和融合:COX18, MFN2;线粒体局限化:MFN2;凋亡基因:BBC3, BCL2, BCL2L1, SOD2, P53.结果表明,骨性关节炎软骨细胞中线粒体发生了明显的能量代谢功能障碍.
배경:연골세포구유수복수손연골조직화유지연골완정성적중요작용.선립체공능장애여세포조망、노화화골관절염적병리과정밀절상관.목적:통과선립체 PCR 심편기술연구골성관절염연골세포중선립체기인적차이표체.방법:수집골성관절염환자급정상성인차화후절지자적관절연골세포,경과세포적제취화배양、RNA 제취화질량검측、mRNA 제취화 cDNA 합성등처리후진행실시정량 PCR 검측.결과여결론:소검측도적84개여선립체상관적대검측기인중유18개선립체기인재골성관절염중발생료현저개변.이골성관절염환자연골세포선립체기인상대정상성인연골세포선립체기인적표체배수표시기인표체적개변정황,기중배수개변화배수조절균대우2적상조기인유 BBC3,BCL2,SLC25A37등15개,배수개변소우0.5화배수조절소우-2적하조기인유 CPT1B,SLC25A16,SLC25A24공3개.18개선립체차이기인공능분류여하:막겁화화전위:BCL2, BCL2L1, TP53, UCP1, UCP3;선립체전운공능:BCL2, BCL2L1, CPT1B, FXC1(TIMM10B), MFN2, STARD3, TP53, UCP1, UCP3;소분자전운공능:SLC25A16, SLC25A2, SLC25A24, SLC25A31, SLC25A37;파단백:FXC1(TIMM10B),MFN2;선립체단백전입:COX18, FXC1(TIMM10B);내막전운:FXC1(TIMM10B), TIMM17B;선립체렬변화융합:COX18, MFN2;선립체국한화:MFN2;조망기인:BBC3, BCL2, BCL2L1, SOD2, P53.결과표명,골성관절염연골세포중선립체발생료명현적능량대사공능장애.
BACKGROUND: Chondrocytes play a significant role in repairing damaged cartilage tissue as wel as in maintaining the integrity of the cartilage. Mitochondria are involved in a large amount of biochemical processes, and mitochondrial impairment has a closed relationship with cel apoptosis, senescence and pathological process of osteoarthritis. OBJECTIVE: To detect the differential expression of mitochondrial genes by using gene chip based mitochondrial gene analyses. METHODS: Articular chondrocytes were col ected from healthy people and osteoarthritis patients, then extracted and cultured fol owed by RNA isolation and quality assessment, mRNA isolation and strand cDNA synthesis. After al , real-time quantitative PCR was performed. RESULTS AND CONCLUSION: Among 84 mitochondrial genes, 18 genes were unambiguously identified as significantly altered in osteoarthritis: 15 of them (BBC3, BCL2, SLC25A37, etc.) were up-regulated at both fold changes and fold regulation > 2, and three of them (CPT1B, SLC25A16, SLC25A24) were down-regulated at fold change < 0.5 and fold regulation < 2. The grouping of 18 functional genes is as fol ows: membrane polarization & potential: BCL2, BCL2L1, TP53, UCP1, UCP3; mitochondrial transport: BCL2, BCL2L1, CPT1B, FXC1 (TIMM10B), MFN2, STARD3, TP53, UCP1, UCP3; smal molecule transport: SLC25A16, SLC25A2, SLC25A24, SLC25A31, SLC25A37; targeting proteins to mitochondria: FXC1 (TIMM10B), MFN2; mitochondrion protein import: COX18, FXC1 (TIMM10B); inner membrane translocation: FXC1 (TIMM10B), TIMM17B;mitochondrial fission & fusion: COX18, MFN2; mitochondrial localization: MFN2; apoptotic genes: BBC3, BCL2, BCL2L1, SOD2, P53. These findings indicate that mitochondrial energy metabolism dysfunction occurs obviously in osteoarthritis chondrocytes.