Expression change and role of sex determining region box transcription factor 2 mRNA and the non-coding RNA in the hepatocyte stem changes during the rat liver regeneration / 解剖学报

Objective To explore the role pathway and pattern of the sex determining region box transcription factor 2 (SOX2) and its mRNA interaction with microRNA(miRNAs, miR) and circular RNA(circRNA) at 0 hour and 2 hours in the rat liver regeneration. Methods The rat 2/3 hepatectomy (partial hepatectomy, PH) model was prepared as described by Higgins, the hepatocytes were isolated according to the method of Smedsrod et al, the expression changes of mRNA, miRNA and circRNA [together named as competing endogenous RNAs(ceRNA)] were detected by the large-scale quantitative detection technology, the interaction network of ceRNA was constructed by Cytoscape 3.2 software, and their correlation in expression and role were analyzed by ceRNA comprehensive analysis. Results It was found that at the 0 hour and 2 hours after PH, the ratio value of SOX2 mRNA shows 1.00±0.09 and 2.15±0.48, miR-3558-3p displays 4.53± 0.10 and 0.81±0.16, circRNA_18404 shows 1.24±0.04 and 11.10±0.57, circRNA_18045 displays 1.97±0.47 and 4.44± 0.23. At the same time, the eight kinds of cell dedifferentiation-related genes AT-rich interaction domain 5A (ARID5A), activating transcription factor 3 (ATF3), BTG anti-proliferation factor 2 (BTG2), etc, which are prometed in expression by SOX2, were down-regulated at 0 h after PH, but the cell differentiation-related genes interferon regulatory factor 6 (IRF6) and somatostatin (SST), which are inhibited in expression by SOX2, were up-regulated at 0 hour after PH. On the other hand, the eight kinds of cell dedifferentiation-related genes ARID5A, ATF3, BTG2, etc, which are promoted in expression by SOX2, were up-regulated at 2 hours after PH, but the cell differentiation-related gene SST, which is inhibited in expression by SOX2, was down-regulated, and IRF6 had no meaningful changes in expression at 2 hours after PH. Conclusion The correlation in expression and role of the miRNA, which are inhibited by circRNA, SOX2, its mRNA is inhibited by miRNA, and the cell stem-related genes, which are regulated by SOX2, are helpful for the hepatocyte to be in differentiation state at 0 hour after PH and to be in stem state at 2 hours after PH.

Expression change and role of Kruppel-like factor 4 mRNA, microRNA-881-3p, circular RNA_20298 and circular RNA_14826 in the hepatocyte apoptosis during the rat liver regeneration / 解剖学报

[ Abstract] Objective To explore the role pathway and pattern of the Kruppel-like factor 4 (KLF4) and its mRNA interaction with microRNA (miRNAs) and circular RNA (circRNAs) at 0 hour and the 120 th hour in the rat liver regeneration. Methods The rat 2 / 3 hepatectomy (partial hepatectomy, PH) model was prepared as described by Higgins, the hepatocytes were isolated according to the method of Smedsrod et al, the expression changes of mRNA, miRNA and circRNA together named as competing endogenous RNA (ceRNA) were detected by the large-scale quantitative detection technology, the interaction network of ceRNA was constructed by Cytoscape 3. 2 software, and their correlation in expression and role were analyzed by ceRNA comprehensive analysis. Results It was found that at the 0 hour and the 120th hour PH, the ratio value of KLF4 mRNA showed 1. 00±0. 16 and 3. 14±0. 27, miR-881-3p displays 18. 30±1. 44 and 0. 47±0. 02, circRNA_20298 indicated 0. 32±0. 10 and 4. 24±0. 22, circRNA_14826 showed 0. 42±0. 13 and 0. 61±0. 08. At the same time, the four kinds of cell apoptosis-related genes adrenoceptor beta 2 (ADRB2), dimethylarginine dimethylaminohydrolase 2 (DDAH2), annexin A5 (ANXA5), ect, which were promoted in expression by KLF4, were down-regulated at 0 hour after PH, but the cell apoptosis-related genes synuclein gamma (SNCG), glutathione-disulfide reductase (GSR), FYVE, RhoGEF and PH domain containing 4 (FGD4), ect, which were inhibited in expression by KLF4, were up-regulated at 0 hour after PH. On the other hand, the cell apoptosis-related genes ANXA5 and thymosin beta 10 (TMSB10), which are promoted in expression by KLF4, were up-regulated at the 120th hour after PH, but the cell apoptosis-related genes chloride intracellular channel 4 (CLIC4) and ataxin 3 (ATXN3), ect, which were inhibited in expression by KLF4, were down-regulated at the 120th hour after PH. Conclusion The correlation in expression and role of the miRNAs, which are inhibited by circRNAs, KLF4, its mRNA is inhibited by miRNAs, and the cell apoptosis-related genes, which are regulated by KLF4, are helpful for the hepatocyte to be in active state 0 hour after PH and to be in apoptotic state 120-hour after PH.

Expression change and role of myeloma cancer gene mRNA and the non-coding RNA in the hepatocyte cycle initiation and termination during the rat liver regeneration / 解剖学报

Objective To explore the role pathway and pattern of the myeloma cancer gene (MYC) and its mRNA interaction with the microRNAs(miRNAs) and circular RNA(circRNAs) at hour 0, hour 6 and hour 72 in the rat liver regeneration. Methods The rat 2/3 hepatectomy (PH) model was prepared as described by Higgins, the hepatocytes were isolated according to the method of Smedsrod et al. The expression changes of mRNA, miRNA and circRNA [together named as competing endogenous RNA (ceRNA)] were detected by the large-scale quantitative detection technology, the interaction network of ceRNA was constructed by Cytoscape 3.2 software, and their correlation in expression and role were analyzed by ceRNA comprehensive analysis. Results It was found that at hour 0 and hour 6 after PH, the ratio value of MYC mRNA showed 0.15±0.03 and 2.36±0.20, miR-134-5p indicated 3.22±0.61 and 0.08±0.02, circRNA_12112 displayed 0.68±0.21 and 13.35±3.53. At the same time, the cell cycle initiation-related genes ras association domain family member 1 (RASSF1), cyclin dependent kinase 2 (CDK2), superoxide dismutase 2 (SOD2), which were promoted in expression by MYC, were down-regulated at hour 0 after PH, but the cell cycle initiation-related genes nestin (NES), RAD21 cohesin complex component (RAD21), CUE domain containing 2 (CUEDC2), which are inhibieted in expression by MYC, had no meaningful express changes at hour 0 after PH. On the other hand, the cell cycle initiation-related gene SOD2, which was promoted in expression by MYC, was up-regulated at hour 6 after PH, but the cell cycle initiation-related genes NES, RAD21, CUEDC2, which are inhibieted in expression by MYC, were down-regulated at hour 6 after PH. In contrary, at hour 72 after PH, the ratio value of MYC mRNA showed 2.36±0.20, miR-880-3p indicated 0.54±0.01, circRNA_09599 displayd 0.54±0.16. At the same time, the cell cycle termination-related gene hepatocyte growth factor (HGF), which is promoted in expression by MYC, was up-regulated 72 hours after PH, the cell cycle termination-related genes MET proto-oncogene receptor tyrosine kinase (MET) and cyclin dependent kinase inhibitor 1A (CDKN1A), which are inhibieted in expression by MYC, were down-regulated 72 hours after PH. Conclusion The correlation in expression and role of the miRNAs, which are inhibited by circRNAs, MYC, its mRNA is inhibited by miRNAs, and the cell cycle initiation-related and cell cycle termination-related genes, which are regulated by MYC, are helpful for the hepatocyte to be in cell cycle initiation state at hour 6 after PH and to be in cell cycle termination state at hour 72 after PH.

Advances in biological functions of miR-429 / 解剖学报

MicroRNAs (miRNAs, miR) are endogenous non-coding single-stranded RNAs with a length of about 22 nucleotides. These RNAs play an important biological function within cells, among which, miR-429 has been proved to play an important role in inhibiting tumor development and tumor progression as well as in cell differentiation and neurological diseases by regulating the expression of different target genes. In this paper, the role of miR-429 and its downstream targeted genes in cell proliferation, apoptosis, migration and invasion is summarized.

Expression profile of circularRNA during rat liver regeneration and its regulatory effect on cell proliferation / 解剖学报

Objective This study aims to investigate the expression profile and regulatory effect on cell proliferation of circular RNA(circRNA) in rat liver regeneration (LR). Methods CircRNA expression profile during rat LR of 114 rats ' regenerating liver which induced by 2/3 partial hepatectomy was detected by high-throughput sequencing. MiRanda and TargetScan were performed to predict their target microRNA (miRNAs) and mRNAs. Gene Ontology (GO) and IP A were used to analyze the physiological activities and signaling pathways they involved. Cytoscape v3. 0. 2 was used to construct the interaction network. Finally, the candidate key circRNAs were selected by the expression pattern combining with the number and function of target miRNAs. Results 20 878 circRNAs were detected during rat LR, among which 560 of them were differentially expressed, and 126 of them could bind to 117 target miRNAs, which were in turn to regulate 6510 downstream target mRNAs. They were involved in cell proliferation, stress response, substance metabolism and transforming growth factor-β (TGF-β), protein kinase A (PKA), Wnt/beta-catenin signaling pathways. 6 differential expressed circRNAs, including circRNA_03651, circRNA_03653, circRNA_04500, circRNA_05865, circRNA_l 1274 and circRNA_ 13559 might play a pivotal role in cell proliferation involved in rat LR by regulating 12 miRNAs and 15 mRNAs. Resultsing they were regarded as the candidate key circRNAs of rat LR. Conclusion 560 circRNAs were differentially expressed in rat LR, among which circRNA_03651, circRNA_03653, circRNA_04500, circRNA_05865, circRNA_l 1274 and circRNA_13559 might play a crucial role on cell proliferation involved in rat LR via 12 miRNAs-15 mRNAs axis.

Targeted metabolomics analysis of amino acid during rat liver regeneration / 解剖学报

Objective To explore the regulation of amino acid metabolism during rat liver regeneration (LR). Methods Rats were randomly divided into 10 groups with 5 rats in each group, including nine partial hepatectomy (PH) groups and one normal control group. Selective reaction monitoring/multi reaction monitoring (SRM/MRM) was employed in the targeted metabolomics identification of 20 amino acids at 10 time points in rat liver regeneration. The change of amino acid content was analyzed by hierarchical clustering. Results Alanine was up-regulated at 30, 36 and 72 hours ; arginine was up-regulated at 6 and 12 hours; aspartic acid was up-regulated at 6, 12 and 36 hours; glutamate was up-regulated at 6, 12, 30, 36, 72 and 120 hours; histidine was up-regulated at 12, 24, 30, 36, 72 and 120 hours; glutamine was up- regulated at 72 hours, and isoleucine was down-regulated at 24 hours. Through hierarchical clustering analysis of amino acids, these amino acids can be grouped into three clusters. Conclusion Many amino acids have changed during the liver regeneration, and throughout the whole process of rat liver regeneration. Changes in amino acids content play an important role in hepatocyte proliferation during the liver regeneration.

Targeted quantitative analysis of energy metabolites in the priming phase during rat liver regeneration / 解剖学报

Objective To investigate the regulation of liver regeneration (LR) by changes in energy metabolites in the initiation phase during rat liver regeneration. Methods Rats were randomly divided into 3 groups with 5 rats in each group, including two partial hepatectomy (PH) groups and one normal control group. Selective reaction monitoring/multiple reaction monitoring (SRM/MRM) was employed in the targeted metabolomics identification of 29 energy metabolites. Ingenuity Pathway Analysis (IPA) was applied for integration analysis, including canonical pathway and molecular interaction network. Results The levels of 3-phospho-D-glycerate, AMP, cyclic AMP, D-fructose 1, 6-bisphosphate, dihydroxyacetome phosphate (DHAP), guanosine monophosphate (GMP), guanosine triphosphate (GTP), nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinueleotide phosphate ( NADP ) significantly increased. The levels of alpha-ketoglutarate, beta-D-fructose 6-phosphate, cis-aconitate, D-glucose 6-phosphate, lactate, NADPH, oxaloacetate and pyruvate dramatically reduced. Through hierarchical clustering analysis of energy metabolisms, these energy metabolisms can be grouped into four clusters. IPA showed that the biomolecular changes in the priming phase of liver regeneration are mainly related to carbohydrate metabolism, cellular growth and proliferation, and organismal development. During the priming phase of liver regeneration, adenosine 5'-monphosphate-activated protein kinase (AMPK), hypoxia- inducible factor la (HIF-la), peroxisome proliferator-activated receptor (PPAR), protein kinase A (PKA) and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathways are involved in energy metabolism, and glycolysis may be the main mode of energy supply. Conclusion The result suggests that the changes of energy matabolites during the initial stage of LR play a regulatory role in live regeneration.

Expression profile of circular RNA and its regulatory effect on autophagy during rat liver regeneration / 解剖学报

Objective Studies have shown that circular RNA (circRNA) is a molecular sponge of microRNA (miRNA) and can be used for the treatment and diagnosis of liver diseases. Therefore, this study aims to further investigate the expression profile of circRNA and its effect on autophagy in rat liver regeneration. Methods Based on the high-throughput sequencing in regenerated liver of 0 hour, 2, 6, 12, 24, 30, 36, 72, 120 and 168 hours after 2/3 hepatectomy in 60 rats, the expression profiles of circRNAs, miRNAs and mRNA were analyzed and their relationship were predicted by miRanda aug 2010 software. Furthermore, according to gene function → miRNA function → circRNA function, circRNAs and miRNAs that regulating autophagy were obtained in liver regeneration and compting endogenous (ceRNA) network was constructed. Results There were 102 circRNAs, 42 miRNAs and 153 mRNAs associated with autophagy involved in liver regeneration. Using bioinformatics analysis, 44 types of circRNAs, 13 types of miRNAs and 17 types of mRNA were found to interact with each other, and their co-expression network was constructed. Conclusion In this study, we screened out the circRNAs, miRNAs and mRNAs related to autophagy in liver regeneration and built the interaction network map.

Expression and role of CCAAT enhancer binding protein ζ mRNA, microRNA-136-3p and four kinds of circular RNAs of hepatocytes during the rat liver regeneration initiation / 解剖学报

Objective To explore the pathways and patterns which the CCAAT enhancer binding protein ζ (CEBPO mRNA, miR-136-3p, rno-Crebrf_0009, rno-Slc38a9_0001, rno-LOCl00362999_0001 and rno- Got1_0001 regulate the hepatocytes in G

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 / 解剖学报

Objective To explore the pathways and patterns which CCAAT/enhancer binding protein β(CEBPβ) mRNA, miR-369-3p and rno-Rmdn2_0006 regulate the hepatocytes in G

Expression and role of CCAAT enhancer binding protein a mRNA, microRNA-144-3p and three kinds of circular RNAs of hepatocytes during the rat liver regeneration initiation / 解剖学报

Objective To explore the pathways and patterns which CCAAT enhancer binding proteina (CEBPα) mRNA, miR-144-3p, rno-LOC100365958_0001, rno-Usp3_0010 and rno-AC241873_0010 regulate the hepatocytes in G

Expression and role of CCAAT enhancer binding protein δ mRNA, microRNA￣3553 and rno￣Acad8_0002 of hepatocytes during the rat liver regeneration initiation / 解剖学报

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 G

Synthesis of five types of nucleobase-alanines and their mechanisms of inhibiting proliferation of rat hepatoma cells RH35 / 解剖学报

Objective Five types of nucleobase-alanines were synthesized to study their cytotoxicity and mechanism. Methods Five types of nucleobase-alanines were synthesized by the method of nucleophilic ring-opening of serine-lactone. MTT assay was used to detect their effects on the viability of rat normal hepatocytes BRL-3A and rat hepatoma cells RH35. Flow cytometry was used to detect their effects of cell cycle and apoptosis on BLR-3A and RH35. Real-time PCR was performed to detect their effects of proliferation/ apoptosis-related gene expression on these two cells. Results Five types of nucleobase-alanines were synthesized, including 1-uracil-L-alanine, 1-thymine-L-alanine, 1-cytosine-L-alanine, 9-adenine-L-alanine and 9-guanine-L-alanine. Among them, 1-cytosine-L-alanine and 9-guanine-L-alanine had significant lower half inhibition concentration(IC

Innate immune cells and liver regeneration / 中国免疫学杂志

Liver regeneration depends on powerful immune system of ownself. TNF-α,IFN-γ,IL-6,IL-12,etc. that secreted by innate immune cells such as macrophages,dendritic cells,NK cells and NKT cells,could act on the hepatocytes and regulate liver regen-eration (LR) through corresponding signaling pathways. This article summarizes the mechanism of different innate immune cells on hep-atocytes,clarifies the recent advances of liver innate immune cellsduring liver regeneration process,lay the foundation for revealing the molecular mechanism of the development of liver regeneration and liver diseases, and for the research and development of new therapeutic methods for liver diseases.