中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (46): 8575-8582.doi: 10.3969/j.issn.2095-4344.2012.46.006

• 骨组织构建 bone tissue construction • 上一篇    下一篇

基于线粒体PCR基因芯片的骨关节炎发病机制

潘剑英，沈 俊，张荣凯，李智富，蔡道章   

  1. 中山大学附属第三医院关节外科，广东省广州市 510630
  • 收稿日期:2012-08-08 修回日期:2012-10-15 出版日期:2012-11-11 发布日期:2012-11-11
  • 通讯作者: 蔡道章，博士，博士生导师，教授，主任医师，中山大学附属第三医院关节外科，广东省广州市 510630 daozhangcai@medmail.com
  • 作者简介:潘剑英★，男，1983年生，汉族，湖南省人，2012年中山大学毕业，硕士，医师，主要从事关节外科临床工作。 storm0132002@163.com

A mitochondrial PCR gene chip based study on the pathogenesis of osteoarthritis

Pan Jian-ying, Shen Jun, Zhang Rong-kai, Li Zhi-fu, Cai Dao-zhang   

  1. Department of Bone and Joint Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China
  • Received:2012-08-08 Revised:2012-10-15 Online:2012-11-11 Published:2012-11-11
  • Contact: Cai Dao-zhang, Doctor, Doctoral supervisor, Professor, Chief physician, Department of Bone and Joint Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China daozhangcai@medmail.com
  • About author:Pan Jian-ying★, Master, Physician, Department of Bone and Joint Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China storm0132002@163.com

PDF

559

被引次数

6

摘要：

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。

关键词: 骨性关节炎, 线粒体PCR芯片, 线粒体, 基因, 凋亡, 发病机制, 代谢, 组织构建

Abstract:

BACKGROUND: Chondrocytes play a significant role in repairing damaged cartilage tissue as well 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 cell 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 collected from healthy people and osteoarthritis patients, then extracted and cultured followed by RNA isolation and quality assessment, mRNA isolation and strand cDNA synthesis. After all, 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 follows: membrane polarization & potential: BCL2, BCL2L1, TP53, UCP1, UCP3; mitochondrial transport: BCL2, BCL2L1, CPT1B, FXC1 (TIMM10B), MFN2, STARD3, TP53, UCP1, UCP3; small 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.

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