动物营养学报 >

2021 , Vol. 33 >Issue 11: 6420 - 6430

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.11.041

分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

柴胡皂苷对热诱导的奶牛乳腺上皮细胞抗氧化能力和热休克蛋白基因表达的影响

  • 胡海涛 ,
  • 孙先枝 ,
  • 黄峰 ,
  • 王青峰 ,
  • 李晶晶 ,
  • 卓钊 ,
  • 范彩云 ,
  • 苏衍菁 ,
  • 程建波
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  • 1. 安徽农业大学动物科技学院, 合肥 230036;
    2. 上海城建职业学院, 上海 201415;
    3. 光明牧业有限公司, 上海 200436
胡海涛(1996-),女,甘肃武威人,硕士研究生,动物营养与饲料科学专业。E-mail:huhaitao711@126.com

收稿日期: 2021-04-16

  网络出版日期: 2021-11-10

基金资助

沪农科推字（2019）第1-2号；国家重点研发计划项目（2016YFD0500503）；安徽省重点研发计划项目（202004a06020006）

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Effects of Saikosaponin on Antioxidant Capacity and Heat Shock Protein Gene Expression in Heat-Induced Bovine Mammary Epithelial Cells

  • HU Haitao ,
  • SUN Xianzhi ,
  • HUANG Feng ,
  • WANG Qingfeng ,
  • LI Jingjing ,
  • ZHUO Zhao ,
  • FAN Caiyun ,
  • SU Yanjing ,
  • CHENG Jianbo
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  • 1. College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;
    2. Shanghai Urban Construction Vocational College, Shanghai 201415, China;
    3. Bright Animal Husbandry Co., Ltd., Shanghai 200436, China

Received date: 2021-04-16

  Online published: 2021-11-10

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摘要

本试验旨在研究柴胡皂苷a （SSa）和柴胡皂苷d （SSd）对热诱导的奶牛乳腺上皮细胞（BMECs）抗氧化能力和热休克蛋白基因表达的影响。以38℃培养的BMECs作为阴性对照组；高温42℃分别培养BMECs 1、4、8、12和24 h，建立热应激模型（阳性对照组）；BMECs培养基中分别添加0.1、1.0 μmol/L SSa和0.01、0.10 μmol/L SSd，38℃培养4 h （SSa和SSd作用细胞时间）后，42℃分别处理1、4、8、12和24 h （处理组）。测定各组BMECs中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和过氧化氢酶（CAT）活性及培养液中丙二醛（MDA）含量，并检测细胞中热休克转录因子-1（HSF-1）和热休克蛋白70（HSP70） mRNA相对表达量。结果表明：与阴性对照组相比，阳性对照组BMECs中SOD和GSH-Px活性显著降低（P<0.05），培养液中MDA含量在热诱导4 h时显著升高（P<0.05），HSF-1和HSP70 mRNA相对表达量显著上调（P<0.05）。与阳性对照组相比，任意浓度的SSa均能够显著提高热诱导1 h时BMECs中SOD活性（P<0.05）；热诱导4、8和24 h时，0.1 μmol/L SSa组BMECs中CAT活性显著提高（P<0.05）；0.01 μmol/L SSd在热诱导1、4、8、12和24 h时能显著提高BMECs中GSH-Px的活性（P<0.05）；但在热诱导12和24 h时，1.0 μmol/L SSa和0.1 μmol/L SSd对BMECs中SOD、GSH-Px和CAT活性无显著改善作用（P>0.05），或者显著降低了以上抗氧化酶的活性（P<0.05），且在热诱导12 h时0.1 μmol/L SSd组培养液中MDA含量显著升高（P<0.05），表明SSa和SSd对BMECs氧化应激的缓解作用存在剂量和时间依赖性。在热诱导1 h时，各处理组BMECs中HSP70 mRNA相对表达量较阳性对照组显著上调（P<0.05）；在热诱导4 h时，0.1 μmol/L SSa处理组BMECs中HSF-1和HSP70 mRNA相对表达量较阳性对照组显著上调（P<0.05）；在热诱导12和24 h时，0.01 μmol/L SSd处理组BMECs中HSP70 mRNA相对表达量较阳性对照组显著上调（P<0.05）。综上所述，42℃热处理可诱导BMECs氧化应激，适宜浓度的SSa和SSd能够缓解热诱导BMECs的氧化应激，其中0.1 μmol/L SSa和0.01 μmol/L SSd对42℃热诱导条件下BMECs的氧化应激有较好的缓解作用，并可提高HSF-1和HSP70基因的表达，保护BMECs免受高温诱导的损伤。

关键词： 奶牛乳腺上皮细胞; 热诱导; 柴胡皂苷; 氧化应激; 热休克蛋白

本文引用格式

胡海涛 , 孙先枝 , 黄峰 , 王青峰 , 李晶晶 , 卓钊 , 范彩云 , 苏衍菁 , 程建波 . 柴胡皂苷对热诱导的奶牛乳腺上皮细胞抗氧化能力和热休克蛋白基因表达的影响[J]. 动物营养学报, 2021 , 33(11) : 6420 -6430 . DOI: 10.3969/j.issn.1006-267x.2021.11.041

Abstract

This experiment was conducted to investigate the effects of saikosaponin-a (SSa) and saikosaponin-d (SSd) on antioxidant capacity and heat shock protein gene expression in heat-induced bovine mammary epithelial cells (BMECs). BMECs were cultured at 38℃ as negative control group; BMECs were cultured at 42℃ for 1, 4, 8, 12 and 24 h to establish a heat stress model (positive control group); BMECs medium was supplemented with 0.1, 1.0 μmol/L SSa and 0.01, 0.10 μmol/L SSd, and then heat induced at 42℃ for 1, 4, 8, 12 and 24 h (treatment groups). The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) in BMECs and the content of malondialdehyde (MDA) in the culture medium were measured of each group, and the mRNA relative expression levels of heat shock transcription factor-1 (HSF-1) and heat shock protein 70 (HSP70) in the cells were also measured. The results showed as follows:compared with the negative control group, the activities of SOD and GSH-Px in BMECs of the positive control group were significantly reduced (P<0.05), the content of MDA in the culture medium was significantly increased at 4 h of heat induction (P<0.05), and the mRNA relative expression levels of HSF-1 and HSP70 were significantly up-regulated (P<0.05). Compared with the positive control group, any concentration of SSa in the culture medium could significantly increase the SOD activity in BMECs at 1 h of heat induction (P<0.05); at 4, 8 and 24 h of heat induction, the CAT activity in BMECs of the 0.1 μmol/L SSa group was significantly increased (P<0.05); 0.01 μmol/L SSd could significantly increase the activity of GSH-Px in BMECs at 1, 4, 8, 12 and 24 h of heat induction (P<0.05); but at 12 and 24 h of heat induction, 1.0 μmol/L SSa and 0.10 μmol/L SSd did not significantly improve the activities of SOD, GSH-Px and CAT in BMECs (P>0.05), or significantly reduced the above antioxidant enzyme activities (P<0.05); the content of MDA in the culture medium of the 0.1 μmol/L SSd group was significantly increased at 12 h of heat induction (P<0.05), indicated that the relieving effect of SSa and SSd on oxidative stress of BMECs presented dose- and time-dependent manner. At 1 h of heat induction, the mRNA relative expression level of HSP70 in BMECs of the treatment groups was significantly upregulated compared with the positive control group (P<0.05); at 4 h of heat induction, the mRNA relative expression levels of HSF-1 and HSP70 in BMECs of the 0.1 μmol/L SSa-treated group was significantly upregulated compared with the positive control group (P<0.05), and at 12 and 24 h of heat induction, the mRNA relative expression level of HSP70 in BMECs of the 0.01 μmol/L SSd group was significantly up-regulated compared with the positive control group (P<0.05). In conclusion, heat treatment at 42℃ induces oxidative stress in BMECs, the proper concentrations of SSa and SSd alleviate heat-induced oxidative stress in BMECs, in which 0.1 μmol/L SSa and 0.01 μmol/L SSd have better alleviating effect on oxidative stress in BMECs, and also increase HSF-1 and HSP70 gene expression in BMECs under 42℃ heat-induced conditions, thus, BMECs are protected from cell damage caused by high temperature induction.

Key words： bovine mammary epithelial cells; heat induction; saikosaponin; oxidative stress; heat shock protein

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