嗜酸乳杆菌抗氧化活性评价及其对小鼠单核巨噬细胞氧化应激的影响  

Evaluation of Antioxidant Activity of Lactobacillus acidophilus andIts Effects on Oxidative Stress in Macrophages of Mice

作　　者：王恒 李建喜[1] 郭志廷[1] 岳聪 张康[1] 仇正英[1] 张凯[1] 王磊[1] 王贵波[1] 王学智[1] 杨孝朴[2] 张景艳[1] WANG Heng;LI Jianxi;GUO Zhiting;YUE Cong;ZHANG Kang;QIU Zhengying;ZHANG Kai;WANG Lei;WANG Guibo;WANG Xuezhi;YANG Xiaopu;ZHANG Jingyan(Gansu China Veterinary Medicine Engineering Technology Research Center,Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences,Lanzhou 730050,China;College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所,甘肃省中兽药工程技术研究中心,兰州730050 [2]甘肃农业大学动物医学院,兰州730070

出　　处：《动物营养学报》2021年第9期5290-5299,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31602101);兰州市人才创新创业项目(2020-RC-147);中国农业科学院兰州畜牧与兽药研究所所级重点任务(CAAS-LMY-02)。

摘　　要：本试验旨在评价嗜酸乳杆菌抗氧化活性及其对小鼠单核巨噬细胞(RAW 264.7细胞)氧化应激的影响。制备嗜酸乳杆菌培养物上清液、完整细胞和胞内提取物,比较其对1,1-二苯基-2-三硝基苯肼(DPPH)、羟自由基和亚油酸的清除能力。筛选抗氧化能力较强的嗜酸乳杆菌培养物上清液,设置4个剂量(5、25、50和125μL)分别作用于RAW 264.7细胞,对照组加入1 mL完全培养液,检测RAW 264.7细胞中一氧化氮(NO)含量、活性氧(ROS)水平以及超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性,并进一步探究其对脂多糖(LPS)诱导的RAW 264.7细胞氧化应激的保护作用。结果表明:1)嗜酸乳杆菌培养物上清液的DPPH、羟自由基和亚油酸清除率均显著高于嗜酸乳杆菌培养物完整细胞和胞内提取物(P<0.05)。2)将嗜酸乳杆菌培养物上清液作用于正常RAW 264.7细胞,与对照组相比,25、50和125μL嗜酸乳杆菌培养物上清液显著提高了RAW 264.7细胞的细胞活力(P<0.05),125μL嗜酸乳杆菌培养物上清液显著提高了RAW 264.7细胞中NO含量(P<0.05),25和50μL嗜酸乳杆菌培养物上清液显著提高了RAW 264.7细胞中SOD活性(P<0.05),125μL嗜酸乳杆菌培养物上清液显著降低了RAW 264.7细胞中GSH-Px活性(P<0.05)。3)将嗜酸乳杆菌培养物上清液作用于LPS诱导的RAW 264.7细胞,与LPS组相比,25、125μL嗜酸乳杆菌培养物上清液显著降低了LPS诱导的RAW 264.7细胞中NO含量(P<0.05),25μL嗜酸乳杆菌培养物上清液显著提高了LPS诱导的RAW 264.7细胞中SOD和GSH-Px活性(P<0.05)。综上所述,嗜酸乳杆菌培养物各组分均具有DPPH、羟自由基和亚油酸清除能力,作为其中抗氧化活性较好的嗜酸乳杆菌培养物上清液可调节LPS诱导引起的RAW 264.7细胞氧化应激。The aim of this study was to investigate the antioxidant activity of Lactobacillus acidophilus and its effects on oxidative stress in macrophages(RAW 264.7 cells)of mice.The Lactobacillus acidophilus culture supernatant,intact cells and intracellular extracts were prepared,and compared the scavenging activities on 1,1-diphenyl-2-picrylhydrazyl(DPPH),hydroxyl radical and linoleic acid.The Lactobacillus acidophilus culture supernatant with high antioxidant capacity was set 4 dose(5,25,50 and 125μL)in RAW 264.7 cells,the control group was added with 1 mL complete culture medium,to detect nitric oxide(NO)content,reactive oxygen species(ROS)level and superoxide dismutase(SOD)and glutathione peroxidase(GSH-PPx)activities in RAW 264.7 cells,and further explored its protective effect on oxidative stress of RAW 264.7 cells induced by lipopolysaccharide(LPS).The results showed as follows:1)the clearance rates of DPPH,hydroxyl free radicals and linoleic acid of Lactobacillus acidophilus culture supernatant were significantly higher than those of Lactobacillus acidophilus culture,intact cells and intracellular extracts(P<0.05).2)When the Lactobacillus acidophilus culture supernatant added in the normal RAW 264.7 cells,compared with control group,the 25,50 and 125μL Lactobacillus acidophilus culture supernatant significantly increased the cell viability of RAW 264.7 cells(P<0.05),the 125μL Lactobacillus acidophilus culture supernatant significantly increased the NO content of RAW 264.7 cells(P<0.05),the 25 and 50μL Lactobacillus acidophilus culture supernatant significantly increased the SOD activity of RAW 264.7 cells(P<0.05),and the 125μL Lactobacillus acidophilus culture supernatant significantly decreased the GSH-Px activity of RAW 264.7 cells(P<0.05).3)When the Lactobacillus acidophilus culture supernatant added in the normal RAW 264.7 cells induced by LPS,compared with the LPS group,the 25 and 125μL Lactobacillus acidophilus culture supernatant significantly decreased the NO content of RAW 264.7 cells induced by L

关 键 词：嗜酸乳杆菌 脂多糖 抗氧化 超氧化物歧化酶 谷胱甘肽过氧化物酶 

分 类 号：S811.3[农业科学—畜牧学]