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袁发浒,胡松,黄丽霞,黄念芳,宋文剑,孙宾莲.基于基因芯片的TNF-α 诱导骨关节炎细胞模型生物信息学分析[J].中国比较医学杂志,2019,29(7):53~60.
基于基因芯片的TNF-α 诱导骨关节炎细胞模型生物信息学分析
Bioinformatics analysis of TNF-α-induced osteoarthritis cell model through microarray analysis of gene expression profiles
投稿时间:2018-12-15  
DOI:10.3969/j. issn. 1671 -7856. 2019. 07. 009
中文关键词:  骨关节炎  转录组  软骨细胞  基因芯片  肿瘤坏死因子α  小鼠
英文关键词:osteoarthritis  transcriptome  chondrocytes  gene chips  tumor necrosis factor-α(TNF-α)  mouse
基金项目:
作者单位E-mail
袁发浒 江汉大学,武汉 430056 yuanfh@ jhun.edu.cn 
胡松 江汉大学,武汉 430056 husongjhun@ qq.com 
黄丽霞 江汉大学,武汉 430056  
黄念芳 江汉大学,武汉 430056  
宋文剑 江汉大学,武汉 430056  
孙宾莲 江汉大学,武汉 430056 binlian17@ jhun.edu.cn 
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中文摘要:
      目的 骨关节炎(osteoarthritis,OA)是最常见的退行性慢性关节疾病,然而其具体的基因机制至今尚不完全清楚,通过基因芯片检测OA 细胞模型的基因表达谱变化,为深入研究OA 发病机制提供更多生物学依据?方法 胰酶结合胶原酶分离获得小鼠原代软骨细胞,以50 ng/ mL 的肿瘤坏死因子α(TNF-α)孵育原代软骨细胞24 h 制备OA 细胞模型?收获细胞提取总RNA 用于基因芯片检测,以表达差异倍数(fold change,FC)>2 且P<0. 01 为条件筛选差异表达基因(differentially expressed genes,DEGs),利用生物信息学软件对差异表达结果进行基因功能分类体系(gene ontology,GO)?KEGG 通路注释分析?结果 原代软骨细胞经TNF-α 处理后,共筛选获得8096 个表达上调DEG 和6413 个表达下调DEG,其中有诸如基质金属蛋白酶?炎性因子?基因凋亡及成骨相关基因等已知的OA 相关的差异表达基因?此外,还有Olfml1 等olfactomedin 超家族成员?Nf1 等未见报道的与OA 相关的基因,尤其发现大量细胞色素超家族成员基因的异常表达,提示线粒体相关功能基因及信号途径可能与OA 进程有重要关联?结论 项目从转录组水平整体分析了TNF-α 诱导的OA 软骨细胞模型基因表达谱变化,为进一步深入探究OA 发病机制提供了新思路?
英文摘要:
      Objective Osteoarthritis (OA) is the most common type of degenerative chronic joint disease, butthe exact genetic mechanisms are still unclear. The aim of this study was to analyze the gene expression profile in an OAcell model detected using a gene chip, and to provide a biological basis for the pathogenesis of OA. Methods Primarymouse chondrocytes were isolated using trypsin combined with collagenase, and the cells were incubated with 50 ng/ mLTNF-α for 24 h. Total RNA was extracted from the harvested cells for gene chip detection to identify differentially expressedgenes (DEGs). A fold change (FC) greater than two and P<0. 01 were the conditions required for each DEG. Biologicalinformation software was used to conduct gene ontology (GO) and KEGG pathway annotation analysis of DEGs. Results After TNF-α treatment, a total of 8096 up-regulated DEGs and 6413 down-regulated DEGs were identified, including genesthat are known to be associated with OA, such as matrix metalloproteinase, inflammatory factors, and apoptosis-related andosteoblast-related genes. In addition, Olfml1 and other olfactomedin superfamily members, Nf1 and other previouslyunreported genes related to OA, were also found. In particular, abnormal expression of a large number of genes related tocytochrome C superfamily members was found, suggesting that mitochondria-related functional genes and signaling pathwaysmay be significantly associated with OA. Conclusions The changes in the gene expression profile in a TNF-α induced OAchondrocyte model at the level of the transcriptome are datected in this study, providing new insights into the pathogenesis of OA.
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