动物营养学报 >

2022 , Vol. 34 >Issue 2: 1276 - 1284

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.02.059

研究论文 RESEARCH PAPER

丁酸钠对脂多糖诱导的奶牛乳腺上皮细胞系炎性损伤的修复作用

  • 李林 ,
  • 宫彬彬 ,
  • 许长锋 ,
  • 曹萌 ,
  • 李建嫄 ,
  • 赵梅 ,
  • 唐伟斌
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  • 1. 邢台学院生物科学与工程学院, 邢台 054001;
    2. 河北医科大学附属邢台人民医院病理科, 邢台 054001
李林(1991-),男,河北隆尧人,讲师,博士,从事动物生物化学研究。E-mail:linl1991@163.com

收稿日期: 2021-06-27

  网络出版日期: 2022-02-15

基金资助

河北省青年自然科学基金（C202010800）

收起

Repair Effects of Sodium Butyrate on Inflammatory Injury of Lipopolysaccharide-Induced Bovine Mammary Epithelial Cell Line

  • LI Lin ,
  • GONG Binbin ,
  • XU Changfeng ,
  • CAO Meng ,
  • LI Jianyuan ,
  • ZHAO Mei ,
  • TANG Weibin
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  • 1. School of Biological Science and Engineering, Xingtai University, Xingtai 054001, China;
    2. Department of Pathology, Xingtai People's Hospital, Hebei Medical University Affiliated Hospital, Xingtai 054001, China

Received date: 2021-06-27

  Online published: 2022-02-15

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摘要

本研究拟通过分析丁酸钠对脂多糖（LPS）诱导的奶牛乳腺上皮细胞系（MAC-T细胞）炎性损伤的修复作用，进一步从体外角度阐述丁酸钠对奶牛乳腺健康的调控机制。在MAC-T细胞中添加不同浓度（0、1、10、100、1 000、10 000 ng/mL）的LPS，检测细胞活力，以确定LPS的适宜浓度，建立细胞氧化损伤模型；并进一步在MAC-T细胞中添加不同浓度（0、2、4、8、16、32 μmol/L）的丁酸钠，检测细胞凋亡率，以确定丁酸钠的适宜浓度。最终选用1 000 ng/mL LPS和16 μmol/L丁酸钠用于本试验。试验分为3组，分别为对照组、LPS处理组和LPS+丁酸钠处理组，分别对其细胞形态、氧化应激指标及凋亡蛋白mRNA表达水平进行检测。结果表明：1）对照组MAC-T细胞呈扁平的无规则形态，贴壁状态良好；而LPS处理组MAC-T细胞核固缩、破裂，并出现大面积死亡脱落现象；LPS+丁酸钠处理组MAC-T细胞边缘清楚，胞内颗粒较少，死亡脱落现象明显减少。2）与对照组相比，LPS处理组MAC-T细胞中超氧化物歧化酶（SOD）活性和总抗氧化力（T-AOC）显著降低（P<0.05），丙二醛（MDA）含量显著升高（P<0.05）；与LPS处理组相比，LPS+丁酸钠处理组MAC-T细胞中SOD活性和T-AOC含量显著升高（P<0.05），MDA含量显著降低（P<0.05）。3）与对照组相比，LPS处理组MAC-T细胞中半胱天冬蛋白酶-3（Caspase-3）、半胱天冬蛋白酶-9（Caspase-9）、B细胞淋巴瘤/白血病-2相关X蛋白（Bax） mRNA表达水平显著或极显著升高（P<0.05或P<0.01），B细胞淋巴瘤/白血病-2（Bcl-2） mRNA表达水平极显著降低（P<0.01）；与LPS处理组相比，LPS+丁酸钠处理组MAC-T细胞中Caspase-3、Caspase-9 mRNA表达水平显著下降（P<0.05），Bcl-2 mRNA表达水平显著升高（P<0.05）。由此可见，丁酸钠对LPS造成的MAC-T细胞氧化损伤起到了一定的修复作用，减少了细胞凋亡的发生。

关键词： 丁酸钠; LPS; MAC-T; 氧化应激; 细胞凋亡

本文引用格式

李林 , 宫彬彬 , 许长锋 , 曹萌 , 李建嫄 , 赵梅 , 唐伟斌 . 丁酸钠对脂多糖诱导的奶牛乳腺上皮细胞系炎性损伤的修复作用[J]. 动物营养学报, 2022 , 34(2) : 1276 -1284 . DOI: 10.3969/j.issn.1006-267x.2022.02.059

Abstract

This study intended to analyze the repair effects of sodium butyrate on inflammatory injury of lipopolysaccharide (LPS)-induced bovine mammary epithelial cell line (MAC-T), and to expound the regulatory mechanism of sodium butyrate on mammary gland health of dairy cows in vitro. The oxidative damage model was established by adding different concentrations (0, 1, 10, 100, 1 000 and 10 000 ng/mL) of LPS into MAC-T cells and determining the cell viability, to determine the appropriate concentration of LPS. In order to determine the appropriate concentration of sodium butyrate, the apoptosis rate of cells was detected by adding different concentrations (0, 2, 4, 8, 16 and 32 μmol/L) of sodium butyrate into MAC-T cells. Finally, 1 000 ng/mL LPS and 16 μmol/L sodium butyrate were selected for this experiment. The experiment was divided into three groups:control group, LPS treatment group and LPS+sodium butyrate treatment group, and the cells morphology, oxidative stress indexes and of apoptosis protein mRNA expression levels were detected. The results showed as follows:1) the cells in the control group showed a flat, irregular shape and were well attached; the cells in the LPS treatment group had pyknosis and rupture of the nuclei, and a large area of death and shedding occurred; the cell edges in the LPS+sodium butyrate treatment group were clear, the intracellular particles were less, and the death and exfoliation were reduced obviously. 2) Compared with the control group, the superoxide dismutase (SOD) activity and total antioxidant stress capacity (T-AOC) in MAC-T cells of LPS treatment group were significantly decreased (P<0.05), and the malondialdehyde (MDA) content was significantly increased (P<0.05); compared with the LPS treatment group, the SOD activity and T-AOC in MAC-T cells of LPS+sodium butyrate treatment group were significantly increased (P<0.05), and the MDA content was significantly decreased (P<0.05). 3) Compared with the control group, the mRNA expression levels of cysteinyl aspartate specific proteinase-3 (Caspase-3), cysteinyl aspartate specific proteinase-9 (Caspase-9) and B-cell lymphoma/leukaemia-2-associated X protein (Bax) in MAC-T cells of LPS+sodium butyrate treatment group were significantly increased (P<0.05 or P<0.01), and the B-cell lymphoma/leukaemia-2 (Bcl-2) mRNA expression level was significantly decreased (P<0.05); compared with the LPS treatment group, the mRNA expression levels of Caspase-3 and Caspase-9 in MAC-T cells of LPS+sodium butyrate treatment group were significantly decreased (P<0.05), and the Bcl-2 mRNA expression level was significantly increased (P<0.05). In conclusion, sodium butyrate plays a certain role in repairing the oxidative damage of MAC-T cells caused by LPS and reduces the occurrence of cell apoptosis.

Key words： sodium butyrate; LPS; MAC-T cells; oxidative stress; cell apoptosis

参考文献

[1] 陈志, 张逸, 路钦越, 等.茶树油对LPS诱导的奶牛乳腺炎的作用及其机制[J].中国农业科学, 2021, 54(14):3124-3133. CHEN Z, ZHANG Y, LU Q Y, et al.Effect and mechanism of tea tree oil on LPS induced mastitis in dairy cows[J].Scientia Agricultura Sinica, 2021, 54(14):3124-3133.(in Chinese)
[2] 朱智, 纪邑奇, 黄文明, 等.丁酸在亚急性瘤胃酸中毒和动物健康方面作用机制的研究进展[J].动物营养学报, 2021, 33(8):4201-4212. ZHU Z, JI Y Q, HUANG W M, et al.Research progress on mechanism of butyrate in subacute ruminal acidosis and animal health[J].Chinese Journal of Animal Nutrition, 2021, 33(8):4201-4212.(in Chinese)
[3] DAI H Y, LIU X X, YAN J Y, et al.Sodium butyrate ameliorates high-concentrate diet-induced inflammation in the rumen epithelium of dairy goats[J].Journal of Agricultural and Food Chemistry, 2017, 65(3):596-604.  
[4] 刘文慧, 阿拉腾珠拉, 马露, 等.丁酸钠与丝兰对大肠杆菌K99攻毒哺乳犊牛生长性能及血清抗氧化指标的影响[J].动物营养学报, 2020, 32(9):4177-4184. LIU W H, A L T Z L, MA L, et al.Effects of sodium butyrate and yucca on growth performance and serum antioxidant indices of sucking calves challenged with Escherichia coli K99[J].Chinese Journal of Animal Nutrition, 2020, 32(9):4177-4184.(in Chinese)
[5] 刘馨忆.包被丁酸钠对肉仔鸡生长性能、免疫功能及肠道组织形态的影响[J].饲料研究, 2020, 43(6):41-44. LIU X Y.Effect of sodium butyrate on growth performance, immune function and intestinal morphology of broilers[J].Feed Research, 2020, 43(6):41-44.(in Chinese)
[6] QAISRANI S N, VAN KRIMPEN M M, KWAKKEL R P, et al.Diet structure, butyric acid, and fermentable carbohydrates influence growth performance, gut morphology, and cecal fermentation characteristics in broilers[J].Poultry Science, 2015, 94(9):2152-2164.  
[7] 王爱华.丁酸钠的生理功能及其在畜禽生产中的应用[J].中国饲料, 2014(24):23-26. WANG A H.Physiologic function of sodium butyrate and its application in livestock and poultry production[J].China Feed, 2014(24):23-26.(in Chinese)
[8] ABDEL-LATIF H M R, ABDEL-TAWWAB M, DAWOOD M A O, et al.Benefits of dietary butyric acid, sodium butyrate, and their protected forms in aquafeeds:a review[J].Reviews in Fisheries Science & Aquaculture, 2020, 28(4):421-448.  
[9] 洪键, 贾逸敏, 赵茹茜.丁酸通过增强肝线粒体功能缓解高脂诱导的小鼠肥胖[J].中国生物化学与分子生物学报, 2017, 33(12):1266-1273. HONG J, JIA Y M, ZHAO R Q.Butyrate alleviates high fat diet-induced obesity through enhancement of mitochondrial function in the liver of mice[J].Chinese Journal of Biochemistry and Molecular Biology, 2017, 33(12):1266-1273.(in Chinese)
[10] MA X M, ZHOU Z H, ZHANG X J, et al.Sodium butyrate modulates gut microbiota and immune response in colorectal cancer liver metastatic mice[J].Cell Biology and Toxicology, 2020, 36(5):509-515.  
[11] ZHANG R Y, ZHU W Y, JIANG L S, et al.Comparative metabolome analysis of ruminal changes in Holstein dairy cows fed low- or high-concentrate diets[J].Metabolomics, 2017, 13(6):74.
[12] DENNIS T S, SUAREZ-MENA F X, HILL T M, et al.Short communication:effect of replacing corn with beet pulp in a high concentrate diet fed to weaned Holstein calves on diet digestibility and growth[J].Journal of Dairy Science, 2018, 101(1):408-412.  
[13] SAMO S P, MALHI M, KACHIWAL A B, et al.Supranutritional selenium level minimizes high concentrate diet-induced epithelial injury by alleviating oxidative stress and apoptosis in colon of goat[J].BMC Veterinary Research, 2020, 16(1):462.
[14] 王来来, 张文文, 张欢敏, 等.丁酸钠对高精料日粮模式下奶山羊乳腺内组胺介导的炎症反应的影响[J].畜牧与兽医, 2017, 49(9):111-116. WANG L L, ZHANG W W, ZHANG H M, et al.Effect of high-concentrate diet feeding and dietary sodium butyrate addition on inflammatory response mediated by histamine in the mammary gland of lactating goats[J].Animal Husbandry & Veterinary Medicine, 2017, 49(9):111-116.(in Chinese)
[15] 张文文, 王来来, 代宏宇, 等.高精料日粮对奶山羊乳腺组织NOD1炎性信号通路的影响[J].畜牧与兽医, 2017, 49(9):101-106. ZHANG W W, WANG L L, DAI H Y, et al.Effect of high-concentrate diet on NOD1 inflammatory signaling pathway in the mammary glands of lactating goats[J].Animal Husbandry & Veterinary Medicine, 2017, 49(9):101-106.(in Chinese)
[16] 苗晋锋, 马海田, 邹思湘, 等.内毒素对山羊乳腺组织中与乳腺炎相关的酶和细胞因子的影响[J].福建农林大学学报(自然科学版), 2007, 36(6):608-613. MIAO J F, MA H T, ZOU S X, et al.Effect of endotoxin on enzymes and cytokines in mammary tissue of goats relating with mastitis[J].Journal of Fujian Agriculture and Forestry University (Natural Science Edition), 2007, 36(6):608-613.(in Chinese)
[17] WU X X, HUANG X L, CHEN R R, et al.Paeoniflorin prevents intestinal barrier disruption and inhibits lipopolysaccharide (LPS)-induced inflammation in caco-2 cell monolayers[J].Inflammation, 2019, 42(6):2215-2225.  
[18] WANG K, LIU X Q, XIAO H, et al.The correlation between inflammatory injury induced by LPS and RAS in EpH4-Ev cells[J].International Immunopharmacology, 2017, 46:23-30.
[19] ZHANG L L, HOU X, SUN L C, et al.Staphylococcus aureus bacteriophage suppresses LPS-induced inflammation in MAC-T bovine mammary epithelial cells[J].Frontiers in Microbiology, 2018, 9:1614.
[20] ZHANG W H, GAO F, ZHU Q F, et al.Dietary sodium butyrate alleviates the oxidative stress induced by corticosterone exposure and improves meat quality in broiler chickens[J].Poultry Science, 2011, 90(11):2592-2599.  
[21] ZHOU D, PAN Q, XIN F Z, et al.Sodium butyrate attenuates high-fat diet-induced steatohepatitis in mice by improving gut microbiota and gastrointestinal barrier[J].World Journal of Gastroenterology, 2017, 23(1):60-75.  
[22] QIU Y Q, MA X Y, YANG X F, et al.Effect of sodium butyrate on cell proliferation and cell cycle in porcine intestinal epithelial (IPEC-J2) cells[J].In Vitro Cellular & Developmental Biology.Animal, 2017, 53(4):304-311.  
[23] WANG Y, BRANICKY R, NOË A, et al.Superoxide dismutases:dual roles in controlling ROS damage and regulating ROS signaling[J].Journal of Cell Biology, 2018, 217(6):1915-1928.  
[24] MOLONEY J N, COTTER T G.ROS signalling in the biology of cancer[J].Seminars in Cell & Developmental Biology, 2018, 80:50-64.
[25] CLÉMENT M V, PERVAIZ S.Reactive oxygen intermediates regulate cellular response to apoptotic stimuli:an hypothesis[J].Free Radical Research, 1999, 30(4):247-252.  
[26] SALIMI V, SHAHSAVARI Z, SAFIZADEH B, et al.Sodium butyrate promotes apoptosis in breast cancer cells through reactive oxygen species (ROS) formation and mitochondrial impairment[J].Lipids in Health and Disease, 2017, 16(1):208.
[27] SUN B, JIA Y M, YANG S, et al.Sodium butyrate protects against high-fat diet-induced oxidative stress in rat liver by promoting expression of nuclear factor E2-related factor 2[J].The British Journal of Nutrition, 2019, 122(4):400-410.  
[28] LUDWIG S, PLESCHKA S, PLANZ O, et al.Ringing the alarm bells:signalling and apoptosis in influenza virus infected cells[J].Cellular Microbiology, 2006, 8(3):375-386.  
[29] ZHOU Z H, XU N B, MATEI N, et al.Sodium butyrate attenuated neuronal apoptosis via GPR41/Gβγ/PI3K/Akt pathway after MCAO in rats[J].Journal of Cerebral Blood Flow and Metabolism, 2021, 41(2):267-281.  
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