急性冷应激大鼠肝脏中novel-miR-133-5p生物学功能预测

Biological function prediction of novel-miR-133-5p in the liver of acute cold stress rat

本研究旨在检测6种novelmicroRNA（novel-miR-1-5P，novelmiR-6-3P，novel-miR-9-3P，novel-miR-25-5p，no-vel-miR-133-5P和novel-miR-287-3p）在急性冷刺激大鼠肝脏中的表达情况，并对novel-miR-133-5P的生物学功能进行分析，预测了novel-miR-133-5P在冷应激大鼠肝脏内的功能。前期冷应激大鼠血清高通量测序试验的结果中发现这6种差异表达的novelmicroRNA。本试验采用了30只12周龄SPF级Wistar雄性大鼠，体质量（350±10）g，将大鼠放置人工气候室预饲7d，预饲温度为（24．0±0．1）℃。随机分成急性冷应激组（CSS）和常温对照组（NTS），对CSS急性冷刺激12h后，冷刺激温度为（4．0±0．1）℃。采取肝脏提取总RNA，应用qRTPCR方法检测6种novelmicroRNA在急性冷刺激大鼠肝脏的表达情况。结果显示，6种novelmicroRNA在急性冷刺激大鼠肝脏均有表达，除novel-miR-9-3P外，其余的都显著升高，novel-miR-133-5P升高的最为明显（P〈0．01）。对novel-miR-1335P进行生物学信息分析发现，novel-miR-133-5P的靶基因有2048个，其中bcl9与wntll肿瘤的发生、增殖和迁移有关；同时wnt11也对正常细胞的增殖、分化和凋亡也有重要影响；Smarcall是一种DNA结合蛋白，可以修复因损伤而停止复制的DNA。结果表明，在大鼠血清中存在的6种novel microRNA同样存在于冷应激大鼠肝脏中，在冷应激状态下novel-miR-133-5P的异常上调可能会对机体的生长发育、免疫调节和遗传等方面有重要的影响。

This study was designed to observe six novel microRNA expression in liver tissue of a cute cold stress rat and predict the function of novel-miR-133-5p in cold stress. Differentially ex- pression of six novel microRNA was found in the serum of cold stress rat by high-throughput sequencing. Thirty SPF Wistar male rats with 12-week-old and weighed （340±20） g were selected. The rats were pre-fed in normal room temperature for seven days,temperature was （24.0±0.1）℃ and then were randomly divided into acute cold stress group（CSS） at （4.0±0.1）℃ and normal control group（NTS） at （24±0.1）℃. After the rats were treated with cold stress for 12 h,total RNA of the liver tissue was extracted and the gene expression of six novel microRNA were as- sayed by qRT-PCR. The results showed that in addition to novet-miR-9-3p,the gene expression of five novel microRNA were significantly improved in acute cold stress group compared with that in normal control group （P〈0.05）, especially the gene expression of novel-miR-133-5p （P〈0.01）. The bioinformatics analysis demonstrated that novel-miR-133-Sp had 2 048 target genes such as bclg,wntll and smarcall, bcl9 and wntll are related with tumor occurrence, proliferation andmigration. Meanwhile,wnt11 also has important effects on normal cell proliferation, differentiation and apoptosis. Smarcall is a DNA-binding protein,responsible for DNA repair. Result suggest that six novel mieroRNA existed in the acute cold stress liver and cold stress-induced up-regulation of novel-miR-133-5p in liver could influence growth development,immunoregulation and hereditary.