南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (02): 213-.

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内毒素血症大鼠心肌组织miRNA差异表达

丁桃,李阳,唐睿珠,张旋,云雨,李健,王殿华   

  • 出版日期:2015-02-20 发布日期:2015-02-20

Differential expression of miRNAs in myocardial tissues of rats with lipopolysaccharideinduced
endotoxemia

  • Online:2015-02-20 Published:2015-02-20

摘要: 目的分析内毒素又称脂多糖(lipopolysaccharide, LPS)血症大鼠心肌组织miRNA表达谱变化,探讨miRNA在内毒素血
症心肌损伤中的作用。方法20只雄性SD大鼠随机分为对照组(n=10)和LPS组(n=10)。LPS组腹腔注射LPS 10 mg/kg,对照
组腹腔注射等量生理盐水。24 h后颈脱臼处死大鼠,取心肌组织。Real-time PCR检测TLR4、TNF-α、IL-1β表达水平,透射电镜
观察其超微结构变化。miRNA芯片筛选心肌组织中差异表达的miRNA,Real-time PCR验证候选miRNA实际表达水平。结
果LPS组大鼠心肌组织TLR4、TNF-α、IL-1β表达明显升高,心肌细胞发生胞质空泡,线粒体水肿、结构破坏等超微结构改变。
芯片技术及Real-time PCR 证实LPS 组大鼠心肌组织中miR-194-3p,miR-344a-3p,miR-465-3p,miR-501-5p,miR-3596c,
miR-185-3p,miR-877显著上调,miR-208b-3p,miR-547-3p,miR-141-3p,miR-28-5p,miR-3585-5p显著下调。结论内毒素血症
大鼠心肌组织中发现的这些差异表达miRNA可能与其心肌损伤发生有关。

Abstract: Objective To analyze the differential miRNA expression profile in the myocardium of rats with lipopolysaccharide
(LPS)-induced endotoxemia and explore the role of miRNA in endotoxin-induced myocardial injury. Methods Twenty male
SD rats received intraperitoneal injection of 10 mg/kg LPS (n=10) or an equivalent amount of saline solution (n=10). At 24 h
after LPS injection, the rats were sacrificed to detect myocardial expressions of TLR4, TNF-α and IL-1β using real-time PCR
and for observing myocardial ultrastructures under transmission electron microscopy. The differentially expressed miRNA in
the myocardium were detected using a miRNA array, and the common differentially expressed miRNAs were selected for
verifying their actual expressions using real-time PCR. Results TLR4, TNF-α and IL-1β were over-activated in the myocardium
of LPS-treated rats, in which mitochondria swelling, structural damaged and cytoplasmic vacuoles were observed. In
LPS-challenged rats, miR-194-3p, miR-344a-3p, miR-465-3p, miR-501-5p, miR-3596c, miR-185-3p, and miR-877 were found
up-regulated significantly, whereas miR-208b-3p, miR-547-3p, miR-141-3p, miR-28-5p, and miR-3585-5p down-regulated in
the myocardium. Conclusion Significant differential expression of the miRNAs occurs in the myocardium of LPS-treated rats,
suggesting their involvement in endotoxin-induced myocardial injury.