[1]周艺琳,王正磊,许媛媛,等.白藜芦醇对乳房链球菌感染引起乳腺细胞氧化应激的影响及机制[J].南京农业大学学报,2020,43(4):698-704.[doi:10.7685/jnau.201909042]
 ZHOU Yilin,WANG Zhenglei,XU Yuanyuan,et al.The effects and mechanism of resveratrol on oxidative stress of mammary cells induced by Streptococcus uberis infection[J].Journal of Nanjing Agricultural University,2020,43(4):698-704.[doi:10.7685/jnau.201909042]
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白藜芦醇对乳房链球菌感染引起乳腺细胞氧化应激的影响及机制()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
43卷
期数:
2020年4期
页码:
698-704
栏目:
动物科学
出版日期:
2020-07-13

文章信息/Info

Title:
The effects and mechanism of resveratrol on oxidative stress of mammary cells induced by Streptococcus uberis infection
作者:
周艺琳1 王正磊1 许媛媛1 韩先干2 Vanhnaseng PHOUTHAPANE3 苗晋锋1
1. 南京农业大学动物医学院, 江苏 南京 210095;
2. 中国农业科学院上海兽医研究所, 上海 200241;
3. 老挝科技部生态与生物技术研究所, 万象 01000
Author(s):
ZHOU Yilin1 WANG Zhenglei1 XU Yuanyuan1 HAN Xiangan2 Vanhnaseng PHOUTHAPANE3 MIAO Jinfeng1
1. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;
2. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China;
3. Biotechnology and Ecology Institute, Ministry of Science and Technology of Laos, Vientiane 01000, Laos
关键词:
小鼠乳腺上皮细胞白藜芦醇乳房链球菌氧化应激Nrf2
Keywords:
mouse mammary epithelial cellresveratrolStreptococcus uberisoxidative stressNrf2
分类号:
S852.4
DOI:
10.7685/jnau.201909042
摘要:
[目的] 本试验旨在探讨白藜芦醇对乳房链球菌(Streptococcus uberis)感染引起乳腺细胞氧化应激的影响及机制。[方法] 先用15 μmol·L-1白藜芦醇(resveratrol)处理小鼠乳腺上皮细胞(EpH4-Ev)4 h,再加入乳房链球菌(MOI=10)共孵育3 h,收集样品;通过生化试剂盒(硝酸还原酶法、比色法、微量法等)、流式细胞术、显微镜观察、Western blot、RT-qPCR等方法检测相关指标的变化。[结果] 乳房链球菌(S.uberis)感染可显著提高EpH4-Ev细胞活性氧(ROS)水平和一氧化氮(NO)、丙二醛(MDA)含量以及诱导型一氧化氮合酶(iNOS)、乳酸脱氢酶(LDH)、N-乙酰-β-D-氨基葡萄糖苷酶(NAGase)、超氧化物歧化酶(SOD)活性,降低总抗氧化能力(T-AOC)水平。白藜芦醇预处理后可显著降低ROS水平、NO含量以及iNOS、LDH、NAGase活性,显著提高T-AOC水平,进一步提高SOD活性。在分子水平上,S.uberis感染显著提高EpH4-Ev细胞血红素加氧酶1(HO-1)、NAD(P)H:醌氧化还原酶1(NQO1)、过氧化氢酶(CAT)、谷胱甘肽S-转移酶(GST)mRNA与核因子E2相关因子2(Nrf2)蛋白的表达水平,而白藜芦醇预处理对此有进一步提升作用。显微镜下观察发现白藜芦醇预处理可以明显改善由S.uberis感染引起的小鼠乳腺上皮细胞皱缩。[结论] 白藜芦醇可以有效缓解乳房链球菌感染引起乳腺细胞的氧化应激,这可能与白藜芦醇激活Nrf2蛋白并促进其下游抗氧化基因表达相关。
Abstract:
[Objectives] The study aimed to explore the effects and mechanism of resveratrol on oxidative stress of mammary cells induced by Streptococcus uberis infection.[Methods] In this study,mice mammary epithelial cells(EpH4-Ev) were treated with 15 μmol·L-1 resveratrol for 4 h,then the S.uberis(MOI=10) was added for 3 h to collect samples. The levels of related indicators were detected by biochemical kit(nitrate reductase,colorimetry,micromethod,et al methods),flow cytometry,microscopy,Western blot,RT-qPCR and the like.[Results] S.uberis infection could significantly increase the level of reactive oxygen species(ROS),the content of nitric oxide(NO),malondialdehyde(MDA),the activity of inducible nitric oxide synthase(iNOS),lactate dehydrogenase(LDH),N-acetyl-β-D-glucosaminidase(NAGase),superoxide dismutase(SOD),and reduce the level of total antioxidant capacity(T-AOC) in EpH4-Ev cells. Resveratrol pretreatment substantially reduced the level of ROS,the content of NO,the activity of iNOS,LDH,NAGase,significantly increased the level of T-AOC and further increased the activity of SOD. At the molecular level,S.uberis infection could obviously improve the mRNA levels of heme oxygenase-1(HO-1),NAD(P) H:quinone oxidoreductase 1(NQO1),catalase(CAT),glutathione S-transferase(GST) and nuclear factor erythroid 2-related factor 2(Nrf2) protein expression in EpH4-Ev cells while resveratrol preconditioning had a further enhancing effect. Microscopic observation showed that resveratrol pretreatment significantly improved the shrinkage of mouse mammary epithelial cells caused by S.uberis infection.[Conclusions] Resveratrol can effectively alleviate the oxidative stress of mammary cells caused by S.uberis infection,which may be related to the activation of Nrf2 and the promotion of downstream antioxidant gene expression by resveratrol.

参考文献/References:

[1] Liu Y J,Chen X Z,Li J. Resveratrol protects against oxidized low-density lipoprotein-induced human umbilical vein endothelial cell apoptosis via inhibition of mitochondrial-derived oxidative stress[J]. Molecular Medicine Reports,2017,15(5):2457-2464.
[2] St?pkowski T M,Kruszewski M K. Molecular cross-talk between the NRF2/KEAP1 signaling pathway,autophagy,and apoptosis[J]. Free Radical Biology and Medicine,2011,50(9):1186-1195.
[3] Zheng L H,Xu Y Y,Lu J Y,et al. Variant innate immune responses of mammary epithelial cells to challenge by Staphylococcus aureus,Escherichia coli and the regulating effect of taurine on these bioprocesses[J]. Free Radical Biology and Medicine,2016,96:166-180.
[4] Li B,Xi P,Wang Z,et al. PI3K/Akt/mTOR signaling pathway participates in Streptococcus uberis-induced inflammation in mammary epithelial cells in concert with the classical TLRs/NF-κB pathway[J]. Veterinary Microbiology,2018,227:103-111.
[5] 代斌. 牛磺酸缓解乳房链球菌感染损伤的机制研究[D]. 南京:南京农业大学,2015. Dai B. The mechanism of taurine attenuated inflamatory injury induced by Streptococcus uberis[D]. Nanjing:Nanjing Agricultural University,2015(in Chinese with English abstract).
[6] 王正磊,周艺琳,周美意,等. 牛磺酸对乳房链球菌感染引起的氧化应激影响及其机制[J]. 南京农业大学学报,2019,42(3):491-498. DOI:10. 7685/jnau.201812007. Wang Z L,Zhou Y L,Zhou M Y,et al. The effects of taurine on oxidative stress induced by Streptococcus uberis infection and its mechanism[J]. Journal of Nanjing Agricultural University,2019,42(3):491-498(in Chinese with English abstract).
[7] Zhang Y,Wang G Y,Wang T R,et al. Nrf2-Keap1 pathway-mediated effects of resveratrol on oxidative stress and apoptosis in hydrogen peroxide-treated rheumatoid arthritis fibroblast-like synoviocytes[J]. Annals of the New York Academy of Sciences,2019,1457(1):166-178.
[8] 金晓露. 白藜芦醇抵御奶牛乳腺上皮细胞氧化应激的作用机制研究[D]. 杭州:浙江大学,2016. Jin X L. Mechanisms underlying the cytoprotective effects of resveratrol against oxidative stress in bovine mammary epithelial cells[D]. Hangzhou:Zhejiang University,2016(in Chinese with English abstract).
[9] Akta? H S,Ozel Y,Ahmad S,et al. Protective effects of resveratrol on hepatic ischemia reperfusion injury in streptozotocin-induced diabetic rats[J]. Molecular and Cellular Biochemistry,2019,460(1/2):217-224.
[10] Drent M,Cobben N A M,Henderson R F,et al. Usefulness of lactate dehydrogenase and its isoenzymes as indicators of lung damage or inflammation[J]. European Respiratory Journal,1996,9(8):1736-1742.
[11] Py?r?l? S,Py?r?l? E. Accuracy of methods using somatic cell count and N-acetyl-β-D-glucosaminidase activity in milk to assess the bacteriological cure of bovine clinical mastitis[J]. Journal of Dairy Science,1997,80(11):2820-2825.
[12] Nair N,Gongora E. Oxidative stress and cardiovascular aging:interaction between NRF-2 and ADMA[J]. Current Cardiology Reviews,2017,13(3):183-188.
[13] Li C,Xu X F,Tao Z H,et al. Resveratrol dimers,nutritional components in grape wine,are selective ROS scavengers and weak Nrf2 activators[J]. Food Chemistry,2015,173:218-223.
[14] Stivala L A,Savio M,Carafoli F,et al. Specific structural determinants are responsible for the antioxidant activity and the cell cycle effects of resveratrol[J]. Journal of Biological Chemistry,2001,276(25):22586-22594.
[15] Harasym J,Oledzki R. Effect of fruit and vegetable antioxidants on total antioxidant capacity of blood plasma[J]. Nutrition,2014,30(5):511-517.
[16] Tsikas D. Assessment of lipid peroxidation by measuring malondialdehyde(MDA)and relatives in biological samples:analytical and biological challenges[J]. Analytical Biochemistry,2017,524:13-30.
[17] 徐靖. 超氧化物歧化酶及其应用的研究进展[J]. 食品工业科技,2013,34(12):387-391. Xu J. Research progress in superoxide dismutase and its application[J]. Science and Technology of Food Industry,2013,34(12):387-391(in Chinese with English abstract).
[18] 胡流芳,王迎,任汝静,等. Keap1-Nrf2/ARE信号通路的抗氧化应激作用及其调控机制[J]. 国际药学研究杂志,2016,43(1):146-152. Hu L F,Wang Y,Ren R J,et al. Anti-oxidative stress actions and regulation mechanisms of Keap1-Nrf2/ARE signal pathway[J]. Journal of International Pharmaceutical Research,2016,43(1):146-152(in Chinese with English abstract).
[19] 阮智杨,陈新,孙雯,等. 奶牛乳房炎病原菌的分离鉴定及药物敏感试验[J]. 畜牧与兽医,2018,50(6):105-109. Ruan Z Y,Chen X,Sun W,et al. Isolation and identification of cow mastitis pathogenic bacteria and disk diffusion test[J]. Animal Husbandry & Veterinary Medicine,2018,50(6):105-109(in Chinese with English abstract).
[20] Mizunoe Y,Kobayashi M,Sudo Y,et al. Trehalose protects against oxidative stress by regulating the Keap1-Nrf2 and autophagy pathways[J]. Redox Biology,2018,15:115-124.
[21] Prakash C,Mishra M,Kumar P,et al. Dehydroepiandrosterone alleviates oxidative stress and apoptosis in iron-induced epilepsy via activation of Nrf2/ARE signal pathway[J]. Brain Research Bulletin,2019,153:181-190.
[22] Zhou Y F,Jin Y C,Yu H,et al. Resveratrol inhibits aflatoxin B1-induced oxidative stress and apoptosis in bovine mammary epithelial cells and is involved the Nrf2 signaling pathway[J]. Toxicon,2019,164:10-15.

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备注/Memo

备注/Memo:
收稿日期:2019-09-25。
基金项目:国家重点研发计划项目(2017YFD0501000,2018YFE0102200)
作者简介:周艺琳,硕士研究生。
通信作者:苗晋锋,教授,博士,主要从事泌乳及动物健康调控研究,E-mail:mjf171647@126.com。
更新日期/Last Update: 1900-01-01