动物营养学报 >

2017 , Vol. 29 >Issue 12: 4398 - 4407

DOI: https://doi.org/10.3969/j.issn.1006-267x.2017.12.020

分子营养 Molecular Nutrition

白藜芦醇通过激活去乙酰化酶1/腺苷一磷酸激活的蛋白激酶信号通路影响牛脂肪细胞凋亡

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  • 1. 山东省农业科学院畜牧兽医研究所, 济南 250100;
    2. 山东省畜禽疫病防治与繁育重点实验室, 济南 250100
成海建(1978-),男,山东菏泽人,助理研究员,硕士,主要从事肉牛营养与遗传育种研究。E-mail:98061107@163.com

收稿日期: 2017-06-11

  网络出版日期: 2017-12-04

基金资助

国家自然科学基金（31402098，31601966）；国家科技支撑计划资助项目（2015BAD03B04）

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Resveratrol Induces Apoptosis of Bovine Adipocytes through Activiate of Sirtuin Type 1/Adenosine Monophosphate Activated Protein Kinase Signaling Pathway

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  • 1. Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan 250100, China;
    2. Shandong Key Lab of Animal Disease Control and Breeding, Jinan 250100, China

Received date: 2017-06-11

  Online published: 2017-12-04

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摘要

本试验旨在研究植物提取物白藜芦醇（RES）对牛皮下脂肪细胞凋亡率以及去乙酰化酶1（SIRT1）/腺苷一磷酸激活的蛋白激酶（AMPK）信号通路关键因子的mRNA和蛋白质表达量的影响。选取18月龄鲁西黄牛的皮下前体脂肪细胞，在细胞分化的第0天，更换为RES浓度分别为0（对照）、100、200和400 μmol/L的培养液处理48 h，每组设3个重复。应用Hoechst 33342染色检测细胞凋亡的形态学变化，流式细胞分析仪检测细胞凋亡率，荧光定量PCR（qPCR）和免疫印迹试验（Western-blot）分别检测SIRT1/AMPK信号通路上的关键因子SIRT1、AMPKα、叉头转录因子1（FoxO1）、B细胞淋巴瘤/白血病-2（Bcl-2）、半胱氨酸天冬氨酸蛋白酶-3（caspase-3）、促凋亡蛋白Bcl-2相关X蛋白（Bax）的mRNA和蛋白质表达量，并利用油红O染色鉴定脂肪细胞。结果表明，与对照组相比：不同浓度RES处理后的牛皮下脂肪细胞凋亡率均极显著增高（P<0.01）；不同浓度RES处理后的SIRT1、AMPKα、caspase-3、Bax的mRNA表达量均显著或极显著提高（P<0.05或P<0.01），Bcl-2的mRNA表达量极显著降低（P<0.01）；不同浓度RES处理后的SIRT1、AMPKα、caspase-3的蛋白质表达量均显著或极显著提高（P<0.05或P<0.01）；200和400 μmol/L RES处理后，FoxO1的mRNA和蛋白质表达量显著或极显著提高（P<0.05或P<0.01），Bax的蛋白质表达量极显著提高（P<0.01），Bcl-2的蛋白质表达量极显著降低（P<0.01）。100 μmol/L RES处理后时，FoxO1的mRNA和蛋白质表达量以及Bcl-2、Bax的蛋白质表达量均与对照组差异不显著（P>0.05）。由此可见，RES通过激活SIRT1/AMPK信号通路，同时激活通路下游的FoxO1，促进了牛皮下脂肪细胞的凋亡，为通过营养调控技术降低牛皮下脂肪沉积提供了一定的理论基础。

关键词： 去乙酰化酶1; 腺苷一磷酸激活的蛋白激酶; 叉头转录因子1; 白藜芦醇; 牛; 脂肪细胞; 凋亡

本文引用格式

成海建, 游伟, 靳青, 刘倚帆, 万发春, 宋恩亮, 刘桂芬, 谭秀文, 张相伦, 刘晓牧 . 白藜芦醇通过激活去乙酰化酶1/腺苷一磷酸激活的蛋白激酶信号通路影响牛脂肪细胞凋亡[J]. 动物营养学报, 2017 , 29(12) : 4398 -4407 . DOI: 10.3969/j.issn.1006-267x.2017.12.020

Abstract

The aim of this study was to explore the effect of plant extracts resveratrol (RES) on the bovine adipocytes apoptosis ratio and key factors mRNA and protein expression in sirtuin type 1 (SIRTl)/adenosine monophosphate activated protein kinase (AMPK) signaling pathway. Bovine subcutaneous preadipocytes of 18-month-old Luxi yellow cattle were selected, on day 0 of cell differentiation, cells were treated with culture medium with 0 (control), 100, 200 and 400 μmol/L RES for 48 h, there were three replicates in each group. Hoechst 33342 staining was used to detect morphological changes of adipocytes apoptosis, the flow cytometry was used to detect the apoptosis ratio, quantitative real-time PCR (qPCR) and Western-blot were selected to detect the gene and protein expressions of SIRTl/AMPK signaling pathway related genes such as SIRT1, AMPKα, forkhead box protein O1 (FoxO1), B cell lymphoma/lewkmia-2 (Bcl-2), cystein-asparate protease-3 (caspase-3), Bcl-2 associated X protein (Bax), and oil red O stain was taken to identify the adipocytes. The results showed that compared with the control group, the adipocyte apoptosis rate after treatment with different concentrations of RES was extremely significantly increased (P<0.01); the mRNA expressions of SIRT1, AMPKα, caspase-3 and Bax after treatment with different concentrations of RES were significantly or extremely significantly increased (P<0.05 or P<0.01), but the Bcl-2 mRNA expression was extremely significantly decreased (P<0.01); the protein expressions of SIRT1, AMPKα and caspase-3 after treatment with different concentrations of RES were significantly or extremely significantly increased (P<0.05 or P<0.01); after treatment with 200 and 400 μmol/L RES, the mRNA and protein expressions of FoxO1 were significantly or extremely significantly increased (P<0.05 or P<0.01), the Bax protein expression was extremely significantly increased (P<0.01), the Bcl-2 protein expression was extremely significantly decreased (P<0.01). After treatment with 100 μmol/L RES, the mRNA and protein expressions of FoxO1 and protein expressions of Bcl-2 and Bax had no significant difference compared with control group (P>0.05). In conclusion, the regulation of apoptosis of bovine subcutaneous adipocytes by RES is through stimulation of AMPK/SIRT1 signaling pathway and activation pathway downstream of FoxO1, which provides a theoretical basis for reducing the caccumulation of subcutaneous fat by nutritional regulation techniques.

Key words： SIRT1; AMPK; FoxO1; resveratrol; bovine; adipocytes; apoptosis

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