Abstract：
BACKGROUND: 2,3,5,4\'-tetrahydroxy-stilbene-2-O-β-D-glucoside (TSG) is the principal bioactive compound contained in Polygonum multiflorum Thunb. (PMT), which is traditionally recorded to possess tonic and anti-aging efficacy.
OBJECTIVE: To identify the TSG-provided promotion on liver regeneration (LR) following partial hepatectomy (PHx) in mice and to explicate its involved mechanism.
METHODS: The promotion of TSG on LR was evaluated by hematoxylin and eosin (H&E), 5-bromodeoxyuridinc (BrdU) and Ki-67 staining, and measuring the level of proliferating cell nuclear antigen (PCNA) and Cyclin D1 in mice with PHx at different time points. Gene Expression Omnibus (GEO, GSE15239) database and the label-free quantitative proteomics from liver of mice at 24 h after PHx were integrated to identify potential involved critical proteins, which were verified by Western-blot, Real-time polymerase chain reaction (RT-PCR), molecular docking and luciferase activity assay. Primary hepatocytes isolated from mice were used to investigate the TSG-provided promotion on proliferation in vitro.
RESULTS: TSG (20 mg/kg) promoted LR in mice after PHx. Results from RNA expression data from clinical samples and proteomic analysis from liver tissues indicated that peroxisome proliferator-activated receptor α (PPARα)-mediated fatty acid metabolism pathway were crucially associated with the TSG-provided promotion on LR. TSG enhanced the nuclear translocation of PPARα and the mRNA expression of a series of PPARα-regulated downstream genes. In addition, TSG lowered hepatic triglyceride (TG) and non-esterified fatty acid (NEFA) amounts and increased hepatic adenosine triphosphate (ATP) level in mice after PHx. TSG up-regulated the transcriptional activity of PPARα in vitro. Next results displayed that TSG promoted cell proliferation as well as ATP level in mice primary hepatocytes, which were abolished when PPARα was suppressed. Meanwhile, the cell viability was also elevated in mice primary hepatocytes treated with ATP.
CONCLUSIONS: Activating PPARα-mediated fatty acid β-oxidation (FAO) pathway led to the production of ATP, which contributed to the TSG-provided promotion on LR after PHx in mice.
OBJECTIVE: To identify the TSG-provided promotion on liver regeneration (LR) following partial hepatectomy (PHx) in mice and to explicate its involved mechanism.
METHODS: The promotion of TSG on LR was evaluated by hematoxylin and eosin (H&E), 5-bromodeoxyuridinc (BrdU) and Ki-67 staining, and measuring the level of proliferating cell nuclear antigen (PCNA) and Cyclin D1 in mice with PHx at different time points. Gene Expression Omnibus (GEO, GSE15239) database and the label-free quantitative proteomics from liver of mice at 24 h after PHx were integrated to identify potential involved critical proteins, which were verified by Western-blot, Real-time polymerase chain reaction (RT-PCR), molecular docking and luciferase activity assay. Primary hepatocytes isolated from mice were used to investigate the TSG-provided promotion on proliferation in vitro.
RESULTS: TSG (20 mg/kg) promoted LR in mice after PHx. Results from RNA expression data from clinical samples and proteomic analysis from liver tissues indicated that peroxisome proliferator-activated receptor α (PPARα)-mediated fatty acid metabolism pathway were crucially associated with the TSG-provided promotion on LR. TSG enhanced the nuclear translocation of PPARα and the mRNA expression of a series of PPARα-regulated downstream genes. In addition, TSG lowered hepatic triglyceride (TG) and non-esterified fatty acid (NEFA) amounts and increased hepatic adenosine triphosphate (ATP) level in mice after PHx. TSG up-regulated the transcriptional activity of PPARα in vitro. Next results displayed that TSG promoted cell proliferation as well as ATP level in mice primary hepatocytes, which were abolished when PPARα was suppressed. Meanwhile, the cell viability was also elevated in mice primary hepatocytes treated with ATP.
CONCLUSIONS: Activating PPARα-mediated fatty acid β-oxidation (FAO) pathway led to the production of ATP, which contributed to the TSG-provided promotion on LR after PHx in mice.
摘要：
背景：2,3,5,4'-四羟基-二苯乙烯-2-O-β-D-葡萄糖苷（TSG）是何首乌中含有的主要生物活性化合物。(PMT),传统上记录具有滋补和抗衰老功效。
目的：确定TSG对小鼠部分肝切除术（PHx）后肝再生（LR）的促进作用，并阐明其相关机制。
方法：用苏木精和伊红（H&E）评价TSG对LR的促进作用，5-溴脱氧尿苷（BrdU）和Ki-67染色，并在不同时间点测量PHx小鼠的增殖细胞核抗原（PCNA）和CyclinD1的水平。基因表达综合（GEO，GSE15239)数据库和来自PHx后24小时小鼠肝脏的无标记定量蛋白质组学被整合以鉴定潜在的涉及的关键蛋白质，通过Western-blot验证,实时聚合酶链反应（RT-PCR），分子对接和荧光素酶活性测定。使用从小鼠分离的原代肝细胞来研究TSG提供的对体外增殖的促进作用。
结果：TSG（20mg/kg）促进PHx后小鼠的LR。来自临床样品的RNA表达数据和来自肝组织的蛋白质组学分析的结果表明，过氧化物酶体增殖物激活受体α（PPARα）介导的脂肪酸代谢途径与TSG提供的LR促进至关重要。TSG增强了PPARα的核易位和一系列PPARα调节下游基因的mRNA表达。此外,TSG降低了PHx后小鼠的肝甘油三酯（TG）和非酯化脂肪酸（NEFA）含量，并增加了肝三磷酸腺苷（ATP）水平。TSG在体外上调PPARα的转录活性。接下来的结果表明，TSG促进小鼠原代肝细胞的细胞增殖以及ATP水平，当PPARα被抑制时被废除。同时,在用ATP处理的小鼠原代肝细胞中,细胞活力也升高.
结论：激活PPARα介导的脂肪酸β-氧化（FAO）途径导致ATP的产生，这有助于TSG在小鼠PHx后对LR的促进。
目的：确定TSG对小鼠部分肝切除术（PHx）后肝再生（LR）的促进作用，并阐明其相关机制。
方法：用苏木精和伊红（H&E）评价TSG对LR的促进作用，5-溴脱氧尿苷（BrdU）和Ki-67染色，并在不同时间点测量PHx小鼠的增殖细胞核抗原（PCNA）和CyclinD1的水平。基因表达综合（GEO，GSE15239)数据库和来自PHx后24小时小鼠肝脏的无标记定量蛋白质组学被整合以鉴定潜在的涉及的关键蛋白质，通过Western-blot验证,实时聚合酶链反应（RT-PCR），分子对接和荧光素酶活性测定。使用从小鼠分离的原代肝细胞来研究TSG提供的对体外增殖的促进作用。
结果：TSG（20mg/kg）促进PHx后小鼠的LR。来自临床样品的RNA表达数据和来自肝组织的蛋白质组学分析的结果表明，过氧化物酶体增殖物激活受体α（PPARα）介导的脂肪酸代谢途径与TSG提供的LR促进至关重要。TSG增强了PPARα的核易位和一系列PPARα调节下游基因的mRNA表达。此外,TSG降低了PHx后小鼠的肝甘油三酯（TG）和非酯化脂肪酸（NEFA）含量，并增加了肝三磷酸腺苷（ATP）水平。TSG在体外上调PPARα的转录活性。接下来的结果表明，TSG促进小鼠原代肝细胞的细胞增殖以及ATP水平，当PPARα被抑制时被废除。同时,在用ATP处理的小鼠原代肝细胞中,细胞活力也升高.
结论：激活PPARα介导的脂肪酸β-氧化（FAO）途径导致ATP的产生，这有助于TSG在小鼠PHx后对LR的促进。
