大鼠肝再生启动阶段肝细胞CCAAT增强子结合蛋白β mRNA、微小RNA-369-3p和rno-Rmdn2_0006的表达和作用  

Expression and role of CCAAT enhancer binding protein β mRNA,microRNA-369-3p and rno-Rmdn2_0006 of hepatocytes during the rat liver regeneration initiation

作　　者：白格 王子慧 宋亚萍 臧夏炎 李亚霏 叶丙雨[1,2] 赵志虎 徐存拴 BAI Ge;WANG Zi-hui;SONG Ya-ping;ZANG Xia-yan;LI Ya-fei;YE Bing-yu;ZHAO Zhi-hu;XU Cun-shuan(College of Life Science,He’nan Normal University,He'nan Xinxiang 453007,China;State Key Laboratory Cultivation Base for Cell Differentiation Regulation,He'nan Xinxiang 453007,China;Institute of Biotechnology,Academy of Military Medical Sciences,Beijing 100071,China)

机构地区：[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 [3]军事医学研究院生物工程研究所,北京100071

出　　处：《解剖学报》2021年第6期913-916,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金(31572270,81771226);国家青年自然科学基金(U1804186,U1404312,31601038,31201093,81200317)。

摘　　要：目的了解大鼠肝再生启动阶段CCAAT增强子结合蛋白β(CEBPβ)mRNA、miR-369-3p和rnoRmdn2_0006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的相关性和作用相关性。结果PH后0 h和6 h时,CEBPβmRNA的比值为1.11±0.11和2.57±0.10,miR-136-3p为0.70±0.22和0.28±0.03,rno-Rmdn2_0006为1.26±0.34和2.62±0.70。CEBPβ促进的G_(0)期相关基因生长停滞和DNA损伤诱导型β(GADD45β)为0.12±0.09和2.50±0.44,细胞周期蛋白依赖性激酶抑制剂1A(CDKN1A)为0.39±0.07和0.93±0.15,抑制的G_(0)期相关基因细胞周期蛋白依赖性激酶2相关蛋白2(CDK2AP2)为2.55±0.42和0.74±0.11,信号转导子和转录激活因子1(STAT1)为2.57±0.13和1.32±0.13。CEBPβ促进的G_(1)期相关基因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,丝裂原活化蛋白激酶14(MAPK14)为1.01±0.15和2.01±0.32,硫氧还蛋白相互作用蛋白(TXNIP)为1.03±0.07和2.50±0.19,抑制的G_(1)期相关基因红细胞衍生核因子2样蛋白2(NFE2L2)为0.66±0.09和0.35±0.05。结论PH后0 h时,CEBPβmRNA未上调,有利于CEBPβ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-Rmdn2_0006和miR-369-3p通过相互作用解除了后者对CEBPβmRNA的抑制,有利于CEBPβ形成,有利于CEBPβ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 CCAAT/enhancer binding proteinβ(CEBPβ)mRNA,miR-369-3p and rno-Rmdn2_0006 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,the 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 hour after PH,the ratio value of CEBPβmRNA showed 1.11±0.11 and 2.57±0.10,miR-136-3p displayed 0.70±0.22 and 0.28±0.03,rno-Rmdn2_0006indicated 1.26±0.34 and 2.62±0.70.The G_(0)phase-related genes promoted by CEBPβwere following,the growth arrest and DNA damage inducible beta(GADD45β)0.12±0.09 and 2.50±0.44,the cyclin dependent kinase inhibitor 1A(CDKN1A)0.39±0.07 and 0.93±0.15,but the G_(0) phase-related genes inbibited by CEBPβwere following,the cyclin dependent kinase 2 associated protein 2(CDK2AP2)2.55±0.42 and 0.74±0.11,the signal transducer and activator of transcription 1(STAT1)2.57±0.13 and 1.32±0.13.The G_(1) phase-related genes promoted by CEBPβwere following,the ETS variant transcription factor 6(ETV6)0.77±0.05 and 2.22±0.68,the hemoglobin oxygenase 1(HMOX1)1.05±0.21 and 4.57±0.88,the mitogen-activated protein kinase 14(MAPK14)1.01±0.15 and 2.01±0.32,the thioredoxin interacting protein(TXNIP)1.03±0.07 and 2.50±0.19,but the G_(1) phase-related gene nuclear factor erythroid 2 like 2(NFE2L2)inbibited by CEBPβ0.66±0.09 and 0.35±0.05.Conclusion CEBPβmRNA is not up-regulated at 0 hour after PH,that is helpful for the expression of the G_(0)phase-related genes inbibited by CEBPβand for the hepatocytes to be in G_(0)phase.On the co

关 键 词：肝再生 G_(0)期肝细胞 G_(1)期肝细胞 CCAAT增强子结合蛋白β 竞争性内源RNA综合分析 生物高通量检测 大鼠 

分 类 号：Q257[生物学—细胞生物学]