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On-line Access: 2016-06-03

Received: 2015-10-31

Revision Accepted: 2016-01-22

Crosschecked: 2016-05-12

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Citations:  Bibtex RefMan EndNote GB/T7714


Feng-hua Liu


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Journal of Zhejiang University SCIENCE B 2016 Vol.17 No.6 P.413-424


Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury

Author(s):  Chen Mei, Sha-sha He, Peng Yin, Lei Xu, Ya-ran Shi, Xiao-hong Yu, An Lyu, Feng-hua Liu, Lin-shu Jiang

Affiliation(s):  Beijing Key Laboratory for Dairy Cow Nutrition, College of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China; more

Corresponding email(s):   liufenghua1209@126.com, jls@bac.edu.cn

Key Words:  Cell-cycle arrest, Cell injury, Heat stress, IEC-6 cell line, Magnolol

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Chen Mei, Sha-sha He, Peng Yin, Lei Xu, Ya-ran Shi, Xiao-hong Yu, An Lyu, Feng-hua Liu, Lin-shu Jiang. Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury[J]. Journal of Zhejiang University Science B, 2016, 17(6): 413-424.

@article{title="Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury",
author="Chen Mei, Sha-sha He, Peng Yin, Lei Xu, Ya-ran Shi, Xiao-hong Yu, An Lyu, Feng-hua Liu, Lin-shu Jiang",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury
%A Chen Mei
%A Sha-sha He
%A Peng Yin
%A Lei Xu
%A Ya-ran Shi
%A Xiao-hong Yu
%A An Lyu
%A Feng-hua Liu
%A Lin-shu Jiang
%J Journal of Zhejiang University SCIENCE B
%V 17
%N 6
%P 413-424
%@ 1673-1581
%D 2016
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1500261

T1 - Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury
A1 - Chen Mei
A1 - Sha-sha He
A1 - Peng Yin
A1 - Lei Xu
A1 - Ya-ran Shi
A1 - Xiao-hong Yu
A1 - An Lyu
A1 - Feng-hua Liu
A1 - Lin-shu Jiang
J0 - Journal of Zhejiang University Science B
VL - 17
IS - 6
SP - 413
EP - 424
%@ 1673-1581
Y1 - 2016
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1500261

Objective: heat stress (HS) is an important environmental stressor that adversely influences livestock during the summer. The aim of this study was to investigate whether magnolol protects against HS-induced intestinal epithelial cell injury. Materials and methods: An intestinal epithelial cell line (IEC-6) was subjected to HS at 42 °C, with and without magnolol pretreatment. cell injury was detected by monitoring lactate dehydrogenase (LDH) release. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay was used to assess cell proliferation and viability, including identifying effective concentrations of magnolol. Flow cytometry confirmed G1-phase cell-cycle arrest and its alleviation by magnolol. Active DNA synthesis was measured by incorporation of nucleic acid 5-ethynyl-2'-deoxyuridine (EdU). G1-phase cell-cycle-related gene expression was assessed by real-time reverse transcription polymerase chain reaction (RT-PCR) and levels of G1-phase-related proteins by Western blotting. Results: HS induced IEC-6 cell injury and decreased cell viability, as demonstrated by data from LDH and MTS assays, respectively. Based on a number of criteria, IEC-6 cells subjected to HS were arrested in the G1 phase of the cell cycle. magnolol pretreatment decreased HS-induced cell injury through relief of this cell-cycle arrest. Conclusions: magnolol pretreatment attenuates HS-induced injury in IEC-6 cells. magnolol is potentially promising as a protective strategy for HS in livestock.


方法:本试验采用大鼠小肠上皮细胞(IEC-6)为研究对象,将其分为五组:对照组(37 °C,5% CO2),热应激组(42 °C,3 h),热应激+厚朴酚低浓度组(5 µmol/L),热应激+厚朴酚中浓度组(10 µmol/L),热应激+厚朴酚高浓度组(20 µmol/L)。采用MTS法复制热应激模型及厚朴酚药物浓度筛选;采用流式细胞术检测热应激造成大鼠小肠上皮细胞细胞周期阻滞及不同浓度厚朴酚缓解细胞周期阻滞情况;利用电子显微镜观察热应激造成的细胞损伤情况;利用荧光电子显微镜观察EdU染色后,热应激对细胞增殖情况的影响及不同浓度厚朴酚的修复作用;采用荧光定量聚合酶链反应(PCR)技术检测热应激对细胞周期基因表达影响及不同浓度厚朴酚对细胞周期基因调节作用,采用蛋白质免疫印迹(Western blot)技术检测热应激对细胞周期蛋白表达的影响及不同浓度厚朴酚对细胞周期蛋白修复作用。
结论:流式细胞术显示热应激造成IEC-6细胞周期阻滞在G1期(96.5%),而三种浓度厚朴酚均可以缓解细胞周期阻滞现象且呈剂量依赖性(5 µmol/L 88.8%,10 µmol/L 81.0%,20 µmol/L 73.5%)。热应激导致阻滞细胞分裂的G1期基因p21p27Rb显著上调(P<0.01),显示细胞周期阻滞,不同浓度厚朴酚下调这三个基因表达且呈剂量依赖性;促进细胞分裂的G1期基因E2F1CDK4cyclin D1表达显著下调(P<0.01),显示细胞周期阻滞不能正常增值,厚朴酚有效上调这三个基因表达水平。热应激导致G1期阻滞细胞分裂蛋白p21及p27上调显著(P<0.01),厚朴酚有效下调p21及p27蛋白表达,且在20 µmol/L浓度时效果最佳;而G1期促进细胞分裂蛋白pRb、E2F1、CDK4和cyclin D1(CCND1)在热应激下表达显著下调(P<0.01),厚朴酚具有一定上调以上四个蛋白的作用,且在20 µmol/L浓度时效果最佳。厚朴酚作为天然成分药物,通过缓解热应激造成的IEC-6细胞G1期细胞周期阻滞,具有缓解热应激造成的细胞损伤能力,有望作为预防畜禽热应激的饲料添加剂。


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