研究论文 RESEARCH PAPER

慢性冷暴露对小鼠肝脏抗氧化功能的影响

  • 刘媛媛 ,
  • 薛念宇 ,
  • 张博熙 ,
  • 李士泽 ,
  • 吕红明
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  • 黑龙江八一农垦大学动物科技学院, 大庆 163319
刘媛媛(1996-),女,新疆石河子人,硕士研究生,研究方向为动物冷应激及其调控机制。E-mail:1457009677@qq.com

收稿日期: 2021-08-10

  网络出版日期: 2022-03-14

基金资助

国家自然科学基金(31972637)

Effects of Chronic Cold Exposure on Hepatic Antioxidant Function in Mice

  • LIU Yuanyuan ,
  • XUE Nianyu ,
  • ZHANG Boxi ,
  • LI Shize ,
  • LYU Hongming
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  • College of Animal Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China

Received date: 2021-08-10

  Online published: 2022-03-14

摘要

本试验旨在探讨慢性冷暴露对小鼠肝脏抗氧化功能的影响。体内慢性冷暴露模型建立:试验将12只3周龄C57BL/6雄性小鼠饲养1周后随机分为2组(对照组和冷暴露组),每组6只。冷暴露组小鼠每日随机放置在4℃环境中3 h,连续4周。冷暴露结束后将所有小鼠同时安乐死,并收集其血液和肝脏。体外冷暴露模型建立:将小鼠肝细胞系AML12细胞随机分为4组(对照组、冷暴露1 h组、冷暴露3 h组、冷暴露6 h组),先在37℃培养箱中培养24 h,然后将冷暴露组细胞置于32℃培养箱中分别亚低温冷刺激0、12、24、36 h,冷刺激结束后将细胞刮取下来进行后续试验。用酶联免疫吸附测定(ELISA)法对小鼠血清中谷胱甘肽(GSH)和丙二醛(MDA)含量进行检测,用Western Blot检测小鼠肝细胞和肝脏组织中抗氧化蛋白硫氧还蛋白-1(Trx-1)和硫氧还蛋白互作蛋白(TXNIP)的表达情况。结果显示:与对照组相比,冷刺激36 h后AML12细胞数量显著下降(P<0.05)。与对照组相比,Trx-1的蛋白表达量在组织和细胞中都显著降低(P<0.05),TXNIP的蛋白表达量显著增加(P<0.05),ELISA检测发现血清GSH含量显著减少(P<0.05),血清MDA含量显著增加(P<0.05)。综上所述,32℃亚低温处理一段时间后会对小鼠肝细胞造成一定的氧化损伤。4℃慢性冷暴露4周后,导致小鼠的抗氧化功能下降。

本文引用格式

刘媛媛 , 薛念宇 , 张博熙 , 李士泽 , 吕红明 . 慢性冷暴露对小鼠肝脏抗氧化功能的影响[J]. 动物营养学报, 2022 , 34(3) : 2023 -2029 . DOI: 10.3969/j.issn.1006-267x.2022.03.060

Abstract

The purpose of this study was to investigate the effects of chronic cold exposure on the antioxidant function of mouse. Establishment of chronic cold exposure model in vivo:twelve 3-week-old C57BL/6 male mice were randomly divided into two groups (control group and cold exposure group) after feeding for one week, with 6 mice in each group. Mice in the cold exposure group were randomly placed at 4℃ for 3 h every day for 4 consecutive weeks. All mice were euthanized at the same time after cold exposure and their blood and liver were collected. In vitro cold exposure model establishment:mice liver cell line AML12 cells were randomly divided into 4 groups (control group, cold exposure group for 1 h, cold exposure group for 3 h, cold exposure group for 6 h). The cells of the cold exposure group were cultured in 37℃ incubator for 24 h, and then placed in 32℃ incubator for 0, 12, 24, 36 h, respectively. After the cold stimulation, the cells were scraped off for follow-up tests. The serum glutathione (GSH)and malondialdehyde (MDA) contents were detected by enzyme linked immunosorbent assay (ELISA). Western Blot was used to measure the expression of thioredoxin-1(Trx-1) and thioredoxin interacting protein (TXNIP) in mouse liver cells and tissues. The results showed that compared with the control group, the number of AML12 cells was significantly decreased after 36 h cold stimulation (P<0.05). Compared with the control group, the protein expression levels of Trx-1 was significantly decreased in both tissues and cells (P<0.05), the protein expression levels of TXNIP was significantly increased (P<0.05). ELISA showed that the content of GSH was significantly decreased (P<0.05) and the content of MDA in serum was significantly increased (P<0.05). In summary, mild low temperature treatment at 32℃ for a period of time can cause certain oxidative damage to mouse liver cells. After 4 weeks of chronic cold exposure at 4℃, the antioxidant function of mice decrease.

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