长链非编码RNA-Co7 Rik的表达特征及其对糖异生的影响

Expression characteristics of long non-coding RNA-Co7Rik and its impacts on gluconeogenesis

目的：揭示长链非编码RNA（long non-coding RNA,LncRNA）Co7Rik的表达特征及其在肝脏糖异生中的潜在作用。方法建立饥饿-再进食动物模型、高脂饮食诱导的肥胖模型, qPCR 法检测LncRNA-Co7Rik表达变化；分离C57BL/6J小鼠不同部位组织,检测其组织表达规律。原代分离小鼠肝脏原代细胞,分别给予二丁酰环腺苷酸（ db-cAMP）、Forskolin、地塞米松、Hydrocortisone等药物刺激,揭示生糖信号对LncRNA-Co7Rik表达的影响。采用核质分离法检测LncRNA-Co7Rik在细胞核/细胞质中的分布特征,进行亚细胞定位。结果动物模型研究显示LncRNA-Co7Rik在饥饿0 h、2 h、5 h、16 h及再进食20 h的表达量分别为1.00±0.61、2.98±0.87、3.41±0.98、6.73±1.21、1.53±0.78（均P〈0.05）,表明LncRNA-Co7Rik的表达水平随饥饿进程显著升高,在再进食后恢复正常水平。且与正常对照组相比,高脂饮食诱导的肥胖小鼠肝脏中LncRNA-Co7Rik的表达明显上调（1.00±0.21对2.92±0.63, P〈0.01）。不同组织定量表明LncRNA-Co7Rik在肝脏组织中特异性表达,表达量是其他组织的3~4倍（P〈0.05）；体外研究发现, LncRNA-Co7Rik的表达受糖异生信号调控,提示其可能参与糖异生过程；亚细胞定位揭示LncRNA-Co7Rik主要分布于细胞核中,提示其可能在基因转录水平发挥作用。结论 LncRNA-Co7Rik是一个受生糖信号调控的肝脏特异性长链非编码RNA,并可能参与糖异生生理、病理过程。

Objective To explore the expression characteristics of long non-coding RNA-Co7Rik and to discuss its potential impacts on hepatic gluconeogenesis. Methods Building fasting-feeding model and high fat diet （ HFD） feeding model to detect the expression level of LncRNA-Co7Rik. Separating different parts of C57BL/6J mice to observe its tissue specificity. Isolating primary hepatocytes and stimulating with dibutyryl cyclic-AMP（ db-cAMP） , Forskolin, dexamethasone, and hydrocortisone to measure the expression changes of LncRNA-Co7Rik. Using cytoplasm and nucleus separate technology to test the distribution characteristics in cell nucleus/cytoplasm of LncRNA-Co7Rik. Results Animal model studies have shown that the relative expression level of LncRNA-Co7Rik in different periods of starvation was 1. 00 ± 0. 61, 2. 98 ± 0. 87, 3. 41 ± 0. 98, 6. 73 ± 1. 21, and 1. 53 ± 0. 78 （ P〈0. 05）, this results indicated that the expression of LncRNA-Co7Rik was significantly increased along with the process of starvation, and returned to normal level after feeding. Meanwhile, LncRNA-Co7Rik was up-regulated in HFD induced obesity model compared with normal group （1. 00 ± 0. 21 vs 2. 92 ± 0. 63, P〈0. 01）. According to qPCR, LncRNA-Co7Rik was specifically expressed in the liver, the expression level was 3-4 folds higher than other tissues （P〈0. 05）. In vitro, LncRNA-Co7Rik was regulated by the signal of gluconeogenesis, which suggested it might be involved in the process of gluconeogenesis. Subcellular localization revealed LncRNA-Co7Rik mainly distributed in the nucleus, suggesting that it may play a role in the gene transcription level. Conclusion LncRNA-Co7Rik is a liver specific LncRNA regulated by glycogenic signal, and may participate in physiological and pathological process of gluconeogenesis.