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

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

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G_(0)期还是G_(1)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,CEBPδmRNA的比值为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G_(0)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G_(1)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,CEBPδmRNA下调,有利于CEBPδ抑制的G_(0)期相关基因表达和肝细胞处于G_(0)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CEBPδmRNA的抑制,有利于CEBPδ形成,有利于CEBPδ促进的G_(1)期相关基因表达和肝细胞处于G_(1)期。

Objective To explore the pathways and patterns which the CCAAT enhancer binding proteinδ(CEBPδ)mRNA,miR-3553 and rno-Acad8_0002 regulate the hepatocytes in G_(0)phase and G_(1)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 CEBPδmRNA showed 0.40±0.08 and 2.15±0.24,miR-3553 displayed 2.53±0.47 and 1.17±0.31,rnoAcad8_0002 indicated 1.24±0.04 and 2.66±0.54.The G_(0)phase-related genes inbibited by CEBPδwere 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 G_(1)phase-related genes promoted by CEBPδwere following,the plasminogen activator urokinase receptor(PLAUR)were 0.27±0.08 and 2.62±0.31,the mitogen-activated protein kinase14(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 CEBPδmRNA is downregulated at 0 hour after PH,that is helpful for the expression of the G_(0) phase-related genes inhibited by CEBPδand for the hepatocytes to be in G_(0)phase.On the contrary,the interaction of miR-3553 and rno-Acad8_0002 leads to CEBPδmRNA to bind with miR-3553,to CEBPδbeing formed probablely,and to the expression of the G_(1) phase-related genes promoted by CEBPβprobablely,and to the hepatocytes being in G_(1) phase at 6 hours after PH.