上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (10): 1194-.doi: 10.3969/j.issn.1674-8115.2010.10.003

• 论著（基础研究） • 上一篇    下一篇

糖尿病小鼠心肌组织microRNA表达谱分析

刁雪红, 申 锷, 胡 兵, 张跃力, 吴作辉, 魏 聪   

  1. 上海交通大学附属第六人民医院超声医学科 上海超声医学研究所心血管病实验室, 上海 200233
  • 出版日期:2010-10-25 发布日期:2010-10-27
  • 通讯作者: 申 锷, 电子信箱: shene1001@hotmail.com。
  • 作者简介:刁雪红（1983—）, 女, 博士生；电子信箱: xuehong819@yahoo.com.cn。
  • 基金资助:

    上海市浦江人才计划（09PJ1408400）

Analysis of microRNA expression profile in cardiac muscle tissues of diabetic mice

DIAO Xue-hong, SHEN E, HU Bing, ZHANG Yue-li, WU Zuo-hui, WEI Cong   

  1. Department of Ultrasound in Medicine, Cardiovascular Disease Laboratory, Shanghai Institute of Ultrasound in Medicine, The Sixth People's Hospital, Shanghai Jiaotong University, Shanghai 200233, China
  • Online:2010-10-25 Published:2010-10-27
  • Supported by:

    Shanghai Pujiang Program, China, 09PJ1408400

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摘要：

目的 观察中晚期糖尿病模型小鼠心肌组织microRNA（miRNA）表达，对差异miRNA调控的靶基因进行初步预测。方法 15只C57小鼠经腹腔一次性注射链脲佐菌素（STZ）建立糖尿病小鼠模型（模型组，n=15），以10只未建模小鼠作为对照组。建模后8周末，超声心动图检测小鼠心脏功能指标，包括左室射血分数（EF）、左室短轴缩短率（FS）和左心室质量指数（LVWI）。动物处死后留取并制备心肌组织标本，HE染色光学显微镜观察心肌细胞形态，并结合相关软件定量分析心肌细胞面积改变；采用微距阵基因芯片技术筛选糖尿病模型小鼠心肌组织差异表达的miRNA，Real-Time PCR验证结果，并对差异miRNA调控的靶基因进行生物信息学分析。结果 建模8周末，超声心动图检测显示，模型组小鼠EF和FS明显小于对照组，LVWI显著大于对照组；差异均有统计学意义（P<0.05）。组织学观察发现，模型组小鼠心肌细胞肥大明显。基因芯片检测并经Real-Time PCR验证发现，模型组小鼠心肌组织中有16个差异表达的miRNA，其中miR-195、miR-199a-3p、miR-700、miR-142-3p、miR-24、miR-21、miR-221、miR-499-3p、miR-208a、miR-705表达上调，miR-29a、miR-1、miR-373、miR-143、miR-20a、miR-220b表达下调。生物信息学分析发现，差异miRNA调控的靶基因与细胞增殖、凋亡、糖代谢及血管生成等生物学功能相关。结论 STZ诱导的中晚期糖尿病模型小鼠心肌组织miRNA表达谱发生明显改变，提示miRNA可能参与糖尿病心肌损伤过程。

关键词: microRNA, 糖尿病, 心肌损伤, 微距阵基因芯片

Abstract:

Objective To observe expression of microRNA (miRNA) in cardiac muscle tissues of advanced diabetic mice and to make an initial prediction of target gene regulated by difference miRNA. Methods Fifteen C57 mice were given single intraperitoneal injection of streptozotocin (STZ) to establish diabetic models (model group). Another 10 normal mice were as control group. At the end of 8 weeks after injection, left ventricular functions, including ejection fraction (EF), fractional shortening (FS), and left ventricular weight index (LVWI), were detected by using echocardiography. The mice were sacrificed for making cardiac muscle tissue samples. Then, cell morphology was observed by HE staining under optical microscope, and change of cell size was analyzed by quantitative software. Differential expressions of miRNA were performed with microarray analysis and further confirmed by quantitative real-time RT-PCR. The target gene regulated by difference miRNA was analyzed with bioinformatics. Results At the end of 8 weeks after injection, EF and FS in model group were significantly lower than those of control group, but LVWI significantly higher (P<0.05). Under histological observation, cardiac hypertrophy showed obviously in model group. There were 16 microRNAs differential expressions detected by microarray analysis in model group, including 10 up-regulated (miR-195, miR-199a-3p, miR-700, miR-142-3p, miR-24, miR-21, miRNA-221, miR-499-3p, miR-208a, and miR-705) and 6 down-regulated (miR-29a, miR-1, miR-373, miR-143, miR-20a, and miR-220b). The bioinformatics analysis indicated that the target gene regulated by miRNA involved in cell proliferation, apoptosis, glycometabolism, and angiogenesis. Conclusion MiRNA expression profile in cardiac muscle tissues of STZ induced advanced diabetic mice has significantly changes, which may contribute toward the process of diabetic cardiomyopathy.

Key words: microRNA, diabetes, cardiomyopathy, microarray