解剖学报

解剖学报 ›› 2021, Vol. 52 ›› Issue (6): 917-920.doi: 10.16098/j.issn.0529-1356.2021.06.012

• 肝再生与细胞周期调控 • 上一篇    下一篇

大鼠肝再生启动阶段肝细胞CCAAT 增强子结合蛋白δ mRNA、微小RNA-3553和rno-Acad8_0002的表达和作用

李亚霏1,2 王子慧1,2 臧夏炎1,2 靳伟1,2 常翠芳1,2 郭建林1,2*  徐存拴1,2*   

  1. 1.河南师范大学生命科学学院，河南 新乡 453007; 2.河南省-科技部共建细胞分化调控国家重点实验室培育基地，河南 新乡 453007
  • 收稿日期:2021-01-04 修回日期:2021-03-12 出版日期:2021-12-06 发布日期:2021-12-06
  • 通讯作者: 郭建林;徐存拴 E-mail:cellkeylab@126.com

Expression and role of CCAAT enhancer binding protein δ mRNA, microRNA-3553 and rno-Acad8_0002 of hepatocytes during the rat liver regeneration initiation

LI Ya-fei1, 2 WANG Zi-hui1, 2 ZANG Xia-yan1, 2 JIN Wei1, 2 CHANG Cui-fang1, 2  GUO Jian-lin1, 2*  XU Cun-shuan1, 2*   

  1. 1. College of Life Science, He’nan Normal University, He’nan Xinxiang 453007, China; 2. State Key Laboratory Cultivation Base for Cell Differentiation Regulation, He’nan Xinxiang 453007, China
  • Received:2021-01-04 Revised:2021-03-12 Online:2021-12-06 Published:2021-12-06
  • Contact: GUO Jian-lin;XU Cun-shuan E-mail:cellkeylab@126.com

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摘要：

目的  了解大鼠肝再生启动阶段CCAAT 增强子结合蛋白 δ（CEBPδ）mRNA、miR3553和rno-Acad8_0002调节肝细胞处于G0期还是G1期的途径和方法。   方法  按Higgins等方法制备大鼠2/3肝切除（PH）模型，按Smedsrod等方法分离肝细胞，用大规模定量分析技术检测大鼠肝再生中肝细胞竞争性内源RNA（ceRNA）表达的变化，用Cytoscape 3.2软件构建ceRNA的相互作用网络,用ceRNA综合分析等方法解析它们表达和作用的相关性。   结果  PH后0 h和 6 h时，CEBPδ mRNA的比0.40±0.08和2.15±0.24，miR-3553为2.53±0.47和1.17±0.31，rno-Acad8_0002为1.24±0.04和2.66±0.54。CEBPδ抑制的G0期相关基因转化生长因子β受体2（TGFBR2）为2.77±0.20和0.79±0.13，磷脂酶A2-IVA（PLA2G4A）为2.56±0.76和0.42±0.13。CEBPδ促进的G1期相关基因纤溶酶原激活剂尿激酶受体（PLAUR）为0.27±0.08和2.62±0.31，丝裂原活化蛋白激酶14（MAPK14）为1.01±0.15和2.01±0.32，ETS变异转录因子6（ETV6）为0.77±0.05和2.22±0.68，血红蛋白加氧酶1（HMOX1）为1.05±0.21和4.57±0.88。   结论  PH后0 h时，CEBPδ mRNA下调，有利于CEBPδ抑制的G0期相关基因表达和肝细胞处于G0期。相反，PH后6 h时，rno-Acad8_0002和miR-3553的相互作用解除了后者对CEBPδ mRNA的抑制，有利于CEBPδ形成，有利于CEBPδ促进的G1期相关基因表达和肝细胞处于G1期。

关键词: 肝再生, G0期肝细胞, G1期肝细胞, CCAAT增强子结合蛋白δ, 竞争性内源RNA综合分析, 生物高通量检测, 大鼠

Abstract:

Objective  To explore the pathways and patterns which the CCAAT enhancer binding protein δ(CEBPδ) mRNA, miR-3553 and rno-Acad8_0002 regulate the hepatocytes in G0 phase and G1 phase during rat liver regeneration (LR).    Methods  The rat 2/3 partial hepatectomy (PH) model was prepared as described by Higgins, the hepatocytes of rat liver right lobes were isolated in 9∶00-11∶00 am according to the method  of Smedsrod et al, a large-scale quantitative detection of competitive endogenous RNA(ceRNAs) was processed by the high-throughput biotechnology, the interaction network of ceRNAs was constructed by Cytoscape 3.2 software, and the correlation in expression and role of ceRNAs was analyzed by ceRNA comprehensive analysis.    Results  It was found that at 0 hour and 6 hours after PH, the ratio value of CEBPδ mRNA showed 0.40±0.08 and 2.15±0.24, miR-3553 displayed 2.53±0.47 and 1.17±0.31, rno-Acad8_0002 indicated 1.24±0.04 and 2.66±0.54. The G0 phase-related genes inbibited by CEBPδ were following, the transforming growth factor beta receptor 2 (TGFBR2) were 2.77±0.20 and 0.79±0.13, the phospholipase A2 group IVA (PLA2G4A) were 2.56±0.76 and 0.42±0.13. The G1 phase-related genes promoted by CEBPδ were following, the plasminogen activator urokinase receptor (PLAUR) were 0.27±0.08 and 2.62±0.31, the mitogen-activated protein kinase 14 (MAPK14) 1.01±0.15 and 2.01±0.32, the ETS variant transcription factor 6 (ETV6) were 0.77±0.05 and 2.22±0.68, the hemoglobin oxygenase 1 (HMOX1) were 1.05±0.21 and 4.57±0.88.   Conclusion  CEBPδ mRNA is down-regulated at 0 hour after PH, that is helpful for the expression of the G0 phase-related genes inhibited by CEBPδ and for the hepatocytes to be in G0 phase. On the contrary, the interaction of miR-3553 and rno-Acad8_0002 leads to CEBPδ mRNA to bind with miR-3553, to CEBPδ being formed probablely, and to the expression of the G1 phase-related genes promoted by CEBPβ probablely, and to the hepatocytes being in G1 phase at 6 hours after PH.

Key words: Liver regeneration, Hepatocyte in G0 phase, Hepatocyte in G1 phase, CCAAT enhancer binding protein δ, Compeititive endogenous RNA comprehensive analysis, Biological high-throughput detection, Rat

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