中国临床解剖学杂志 ›› 2015, Vol. 33 ›› Issue (2): 171-175.doi: 10.13418/j.issn.1001-165x.2015

• 实验研究 • 上一篇    下一篇

急性肌损伤炎症反应及其转录分析

杨同群1,2,   张谦1,   耿喜林1,   胡旭昌1,   丁明聪1,   黄美贤2,   廖华2??   

  1. 1. 兰州大学第二临床医学院骨科, 兰州 730030;   2. 南方医科大学解剖学教研室  广东省组织构建与检测重点实验室, 广州  510515
  • 收稿日期:2014-08-11 出版日期:2015-03-25 发布日期:2015-04-21
  • 通讯作者: 廖华,教授,博士生导师,E-mail:hua-liao@163.com E-mail:yangtongqun609@163.com
  • 作者简介:杨同群(1968-),男, 河北人,副主任医师,研究方向:骨科创伤,Tel:13919195845
  • 基金资助:

    国家自然科学基金(81171724, 81371924)

Transcriptomic analysis of gene alteration related to the regulation of inflammation/immune responses after acute myoinjury

YANG Tong-qun 1,2,   ZHANG Qian 1,   GENG Xi-lin 1,   HU Xu-chang 1,   DING Ming-cong 1,  HUANG Mei-xian 2,  LIAO Hua 2   

  1. 1.Department of Orthopedics, Lanzhou University Second Hospital, Lanzhou 730030, China;  2. Department of Anatomy, Key Laboratory of Construction and Detection of Guangdong Province, Southern Medical University, Guangzhou 510515, China
  • Received:2014-08-11 Online:2015-03-25 Published:2015-04-21

摘要:

目的 RNA芯片转录分析骨骼肌损伤前后炎症/免疫相关基因的表达改变,为肌损伤治疗、肌组织炎性肿胀的预防和处理提供相关的分子信息。 方法   机械挤压法制备B6小鼠腓肠肌损伤模型, 组化及免疫荧光检测肌损伤的程度和过程。RNA芯片转录检测、对比分析肌损伤前后炎症/免疫相关基因的表达改变。 结果   持续2次的挤压处理导致严重的腓肠肌损伤,肌组织肿胀、炎性渗出明显、肌纤维坏死(2~4 d)并逐渐出现再生的中央核细胞(7~10 d)。芯片检测发现在39429个检测探针中,3494个基因因骨骼肌损伤而呈现显著的表达水平改变(P<0.01),319个基因的表达改变超过10倍。富集率最高的gene ontology (GO)主要涉及免疫过程、防御反应和免疫反应。大部分基因在损伤后7~10 d趋于正常水平,41个基因在肌损伤后期仍持续上调,于伤后7 d或10 d达最高水平。 结论   急性肌损伤炎症反应具有瞬时性,属于自限性炎症。再生期持续上调的基因可能参与肌损伤后期的抗炎效应,以避免持续、反复、破环性的骨骼肌炎症。

关键词: 骨骼肌损伤,  , 转录, 基因, 炎症

Abstract:

Objective    To follow change of gene expression over time in crushed muscles of B6 mice using transcriptomic analysis.   Methods   Crush induced myoinjury was performed on gastrocnemius muscle of wide B6 mice. H&E and immunofluorecence staining were used to evaluate the necrosis and regeneration of damaged muscle. A mouse one color GE 4×44K G4846A, V2 microarray kit was used to perform global transcriptal expression analysis using muscle total RNA samples. Feature Extraction Software was used for the analysis of probe signals.  Results   Our microarray data identified 3494 genes (among 39429 probes) differentially expressed in crushed gastrocnemius muscle compared to normal muscle, and about 10% (319 probes) of these genes differed more than ten-fold in expression levels. In general, most of genes encoding proteins with important roles in muscle inflammation and immune up-regulated at day 2 and 4 postinjury, and then rapidly down-regulated and return to normal at day 10, which confirmed the effective but brief inflammatory response after acute myoinjury. 41 inflammatory/immune associated-genes persistently up-regulated at the day 7 and 10 after crush damage, which involved in many anti-inflammatory cytokines, chemokines and growth factor-related genes.  Conclusions   Transcriptome analysis indicates transient behavior of crush-induced muscle immune response. The persistently up-regulated genes maybe have anti-inflammatory effects during the muscle repair.

Key words: Crush, Inflammation, Genes, Transcriptome microarray analysis

中图分类号: