目的探讨大鼠肝再生中肝组织mRNA和微RNA（miRNA）表达变化与肝细胞增殖的关联，及肝再生的分子网络调控规律。方法按Higgins等方法制备大鼠2/3肝切除（PH）模型，于术后0、2、6、12、24、30、36、72、120、168 h等10个时间点取肝右叶，除0点其余9个时间点每个时间点6只大鼠，对照组6只大鼠，用生物芯片定量检测各时间点肝组织的mRNA和miRNA信号值。以PH后0 h的3个信号值为对照，分别除以其他时间点相应mRNA和miRNA的信号值，得到它们的比值。用要素累加法筛选肝再生中肝细胞G₀/G₁过渡期（PH后2、6 h），肝细胞增殖期（PH后2、6、12、24、30 和36 h），G₁/G₀过渡期（PH后72、120和168 h）的有意义、有差异、相关、关键mRNA和miRNA。用生物信息学方法分析关键mRNA和miRNA的作用、相互作用和表达相关性。结果用生物芯片检测出大鼠肝再生10个时间点中共13 927种mRNA和1166种miRNA，用要素累加法从这些mRNA和miRNA中筛选出946种关键mRNA和2种关键miRNA novel701_mature与大鼠（rno）-miR-196a-5p。生物信息学分析显示，novel701_mature与肿瘤坏死因子（TNF）受体超家族成员12A（TNFRSF12A）mRNA相互作用，rno-miR-196a-5p与水通道蛋白4（AQP4）、BTB结构域和CNC同源物1（BACH1）、CD244、同源盒B8（HOXB8）、神经接头蛋白1（NRXN1）、表皮桥粒蛋白（PPL）和TSC复合物亚单位1（TSC1）等的mRNA相互作用。其中，上述两种miRNA在肝细胞G₀/G₁过渡期和增殖期表达上调，G₁/G₀过渡期下调，它们相互作用的上述8种关键mRNA在上述肝细胞增殖的3个时期均表达上调。结论筛选的2种关键miRNA与8种靶mRNA呈现表达相关性，通过相互作用调节大鼠肝再生中肝细胞增殖和肝再生进程。

Objective To elucidate the association between changes in mRNA and microRNA(miRNA) expression and reveal the regulatory patterns in hepatocyte proliferation during rat liver regeneration. MethodsA two-thirds partial hepatectomy (PH) model was established in rats using the Higgins method. Liver right lobe samples were collected at 10 time points (0, 2, 6, 12, 24, 30, 36, 72, 120, and 168 hours) after PH. Except for 0 time point, there were 6 rats in each of the other 9 time points, with 6 rats in the control group. mRNA and miRNA signal values in the liver tissue at each time point were quantitatively detected using microarrays. The signal values of mRNA and miRNA on 0 hour post-PH (three replicates) served as controls. The ratio values of mRNA and miRNA at other time points were calculated by dividing their signal values by the corresponding controls. The element accumulation method was employed to screen biologically significant, differentially expressed, related, and key mRNAs and miRNAs during specific phases of liver regeneration, the hepatocyte G₀/G₁ transition phase (including 2 and 6 hours post-PH), the proliferation phase (including 2, 6, 12, 24, 30, and 36 hours post-PH), and the G₁/G₀ transition phase (including 72, 120, and 168 hours post-PH). Bioinformatics method were used to analyze the functions, interactions, and expression correlations of the identified key mRNAs and miRNAs. ResultsA total of 13 927 mRNAs and 1166 miRNAs were detected across 10 time points during rat liver regeneration using biochip technology. By applying the element accumulation method, 946 key mRNAs and two key miRNAs novel701_mature and rattus morvegicus(rno)-miR-196a-5p were identified. Bioinformatics analysis revealed that novel701_mature interacted with TNFRSF12A mRNA, and rno-miR-196a-5p interacted with mRNAs of aquaporin 4(AQP4), BTB damin and CNC domain and CNC homolog 1(BACH1), CD244, homeo box B8(HOXB8), neurexin 1(NRXN1), periplacin(PPL), and TSC complex subunit 1(TSC1). Furthermore, the expression levels of these two miRNAs were up-regulated during the G₀/G₁ transition and proliferation phases, but down-regulated during the G₁/G₀ transition in hepatocytes. Conversely, the expression levels of the eight key mRNAs that interacted with these two miRNAs were up-regulated across all three phases (G₀/G₁ transition, proliferation, and G₁/G₀ transition) of hepatocyte proliferation. ConclusionThe two key miRNAs show expression correlation with identified eight target mRNAs and regulat hepatocyte proliferation and liver regeneration processes.