动物营养学报 >

2020 , Vol. 32 >Issue 9: 4327 - 4336

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.09.043

分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

单宁酸诱导猪肾细胞氧化损伤和凋亡

  • 王吉 ,
  • 张琳玉 ,
  • 刘翔燕 ,
  • 肖海思 ,
  • 李鑫 ,
  • 严思思 ,
  • 袁志航 ,
  • 梁曾恩妮 ,
  • 文利新 ,
  • 邬静
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  • 1. 湖南农业大学动物医学院, 长沙 410128;
    2. 长沙绿叶生物科技有限公司, 长沙 410100;
    3. 湖南省农业科学院农产品加工研究所, 长沙 410125;
    4. 湖南畜禽安全生产协同创新中心, 长沙 410128
王吉(1990-),男,湖南邵阳人,博士研究生,研究方向为营养代谢与动物保健。E-mail:wangjics@163.com

收稿日期: 2020-03-19

  网络出版日期: 2020-09-17

基金资助

国家重点研发计划资助项目（2016YFD0501209）

收起

Tannic Acid Induces Oxidative Damage and Apoptosis in Pig Kidney Cells

  • WANG Ji ,
  • ZHANG Linyu ,
  • LIU Xiangyan ,
  • XIAO Haisi ,
  • LI Xin ,
  • YAN Sisi ,
  • YUAN Zhihang ,
  • LIANG Zengenni ,
  • WEN Lixin ,
  • WU Jing
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  • 1. Faculty of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China;
    2. Changsha Lvye Biotechnology Co., Ltd., Changsha 410100, China;
    3. Hunan Agricultural Product Processing Institute, Hunan Academy of Agricultural Sciences, Changsha 410125, China;
    4. Hunan Collaborative Innovation Center of Animal Production Safety, Changsha 410128, China

Received date: 2020-03-19

  Online published: 2020-09-17

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摘要

本试验对单宁酸（TA）介导的动物肾细胞毒性的分子机制展开研究，旨在为消减和利用饲料中的抗营养因子提供试验依据。采用猪肾细胞系PK-15细胞，以不同浓度[0（空白对照）、10、20、30 μmol/L]TA处理PK-15细胞24 h后，测定细胞活性、乳酸脱氢酶（LDH）活性、氧化损伤指标、线粒体膜电位（MMP）、凋亡相关蛋白表达量、细胞周期。结果显示：TA剂量依赖性地降低PK-15细胞的活力。与空白对照相比，30 μmol/L TA极显著增加了培养基中LDH的活性（P<0.01）；极显著升高了PK-15细胞中丙二醛（MDA）含量（P<0.01），显著或极显著降低了谷胱甘肽（GSH）含量（P<0.01）以及谷胱甘肽过氧化物酶（GSH-Px）（P<0.01）、超氧化物歧化酶（SOD）（P<0.01）和过氧化氢酶（CAT）（P<0.05）活性，极显著增加了活性氧（ROS）含量（P<0.01）；极显著降低了MMP（P<0.01）。与空白对照相比，30 μmol/L TA可使Bcl-2相关的X蛋白（Bax）/B细胞淋巴瘤-2（Bcl-2）（P<0.01）、剪切体-半胱天冬酶-3（cleaved-Caspase-3）/前体-半胱天冬酶-3（pro-Caspase-3）（P<0.01）和剪切体-半胱天冬酶-9（cleaved-Caspase-3）/前体-半胱天冬酶-9（pro-Caspase-9）（P<0.05）显著或极显著增加；此外，30 μmol/L TA还可诱导PK-15细胞在S期积累，发生S期阻滞。由此得出，高浓度的TA诱导PK-15细胞的氧化损伤和细胞内ROS介导的线粒体途径凋亡，并导致了细胞的S期阻滞。

关键词： 单宁酸; PK-15细胞; 氧化损伤; 细胞凋亡; 抗营养因子

本文引用格式

王吉 , 张琳玉 , 刘翔燕 , 肖海思 , 李鑫 , 严思思 , 袁志航 , 梁曾恩妮 , 文利新 , 邬静 . 单宁酸诱导猪肾细胞氧化损伤和凋亡[J]. 动物营养学报, 2020 , 32(9) : 4327 -4336 . DOI: 10.3969/j.issn.1006-267x.2020.09.043

Abstract

The aim of this study was to explore the molecular mechanism of tannic acid (TA) mediated renal cytotoxicity in animals, and to provide experimental basis for the reduction and utilization of antinutritional factors in feed. Pig kidney cell line PK-15 cells were treated with different concentrations of TA[0 (blank control), 10, 20 and 30 μmol/L] for 24 h. Cell viability, lactate dehydrogenase (LDH) activity, oxidative damage parameters, mitochondrial membrane potential (MMP), apoptosis related protein expression and cell cycle were measured. The results showed that TA decreased the viability of PK-15 cells in a dose-dependent manner. Compared with the blank control, the treatment of 30 μmol/L TA extremely significantly increased the activity of LDH in the medium (P<0.01); extremely significantly increased malondialdehyde (MDA) content of PK-15 cells (P<0.01), significantly or extremely significantly reduced the content of glutathione (GSH) (P<0.01) and the activities of glutathione peroxidase (GSH-Px) (P<0.01), superoxide dismutase (SOD) (P<0.01) and catalase (CAT) (P<0.05), and extremely significantly increased the content of reactive oxygen species (ROS) (P<0.01); extremely significantly reduced the MMP of PK-15 cells (P<0.01). The treatment of 30 μmol/L TA could extremely significantly or significantly increase B cell lymphoma-2 (Bcl-2) associated X protein (Bax)/Bcl-2 (P<0.01), cleaved-Caspase-3/pro-Caspase-3 (P<0.01) and cleaved-Caspase-9/pro-Caspase-9 (P<0.05). In addition, the treatment of 30 μmol/L TA could induce the accumulation of PK-15 cells in S-phase, leading to S-phase arrest. It is concluded that high concentrations of TA induce oxidative damage and intracellular ROS-mediated mitochondrial pathway apoptosis in PK-15 cells, as well as leading to S-phase arrest of the cells.

Key words： tannic acid; PK-15 cells; oxidative damage; cell apoptosis; antinutritional factor

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