EXPERIMENTAL METHOD AND ANIMAL

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

  • WANG Heng ,
  • LI Jianxi ,
  • GUO Zhiting ,
  • YUE Cong ,
  • ZHANG Kang ,
  • QIU Zhengying ,
  • ZHANG Kai ,
  • WANG Lei ,
  • WANG Guibo ,
  • WANG Xuezhi ,
  • YANG Xiaopu ,
  • ZHANG Jingyan
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  • 1. Gansu China Veterinary Medicine Engineering Technology Research Center, Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences, Lanzhou 730050, China;
    2. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China

Received date: 2021-03-07

  Online published: 2021-09-18

Abstract

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 125 μL) 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 125 μL Lactobacillus acidophilus culture supernatant significantly increased the cell viability of RAW 264.7 cells (P<0.05), the 125 μL Lactobacillus acidophilus culture supernatant significantly increased the NO content of RAW 264.7 cells (P<0.05), the 25 and 50 μL Lactobacillus acidophilus culture supernatant significantly increased the SOD activity of RAW 264.7 cells (P<0.05), and the 125 μL 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 125 μL Lactobacillus acidophilus culture supernatant significantly decreased the NO content of RAW 264.7 cells induced by LPS (P<0.05), and the 25 μL Lactobacillus acidophilus culture supernatant significantly increased the activities of SOD and GSH-Px of RAW 264.7 cells induced by LPS (P<0.05). In conclusion, all parts of Lactobacillus acidophilus culture have the scavenging activities on DPPH, hydroxyl radical and linoleic acid, and the Lactobacillus acidophilus culture supernatant with superior activity on antioxidant activity can regulate the oxidative stress of RAW 264.7 cells induced by LPS.

Cite this article

WANG Heng , LI Jianxi , GUO Zhiting , YUE Cong , ZHANG Kang , QIU Zhengying , ZHANG Kai , WANG Lei , WANG Guibo , WANG Xuezhi , YANG Xiaopu , ZHANG Jingyan . Evaluation of Antioxidant Activity of Lactobacillus acidophilus and Its Effects on Oxidative Stress in Macrophages of Mice[J]. Chinese Journal of Animal Nutrition, 2021 , 33(9) : 5290 -5299 . DOI: 10.3969/j.issn.1006-267x.2021.09.048

References

[1] HALLIWELL B.Free radicals in medicine and biology[J].The Lancet,1999,344:721-724.
[2] MARTARELLI D,VERDENELLI M C,SCURI S,et al.Effect of a probiotic intake on oxidant and antioxidant parameters in plasma of athletes during intense exercise training[J].Current Microbiology,2011,62(6):1689-1696.  
[3] MAZZOLI A,DONADIO G,LANZILLI M,et al.Bacillus megaterium SF185 spores exert protective effects against oxidative stress in vivo and in vitro[J].Scientific Reports,2019,9(1):12082.
[4] RAGUL K,KANDASAMY S,DEVI P B,et al.Evaluation of functional properties of potential probiotic isolates from fermented brine pickle[J].Food Chemistry,2020,311:126057.
[5] KUDA T,KAWAHARA M,NEMOTO M,et al.In vitro antioxidant and anti-inflammation properties of lactic acid bacteria isolated from fish intestines and fermented fish from the Sanriku Satoumi region in Japan[J].Food Research International,2014,64:248-255.
[6] LI S Y,ZHAO Y J,ZHANG L,et al.Antioxidant activity of Lactobacillus plantarum strains isolated from traditional Chinese fermented foods[J].Food Chemistry,2012,135(3):1914-1919.  
[7] 李军亮,杨奇慧,谭北平,等.嗜酸乳杆菌对斜带石斑鱼幼鱼生长、消化性能和抗病力的影响[J].水生生物学报,2019,43(5):992-1000. LI J L,YANG Q H,TAN B P,et al.Effects of Lactobacillus acidophilus (GIM:1.730) on growth,digestive capacity,disease resistance of juvenile,Epinephelus coioides[J].Acta Hydrobiologica Sinica,2019,43(5):992-1000.(in Chinese)
[8] 张丹丹.S-层蛋白介导嗜酸乳杆菌对肠道炎症的干预作用及其免疫调节功能[D].硕士学位论文.南京:南京师范大学,2016. ZHANG D D.S-layer proteins mediate the effects of Lactobacillus acidophilus on intestinal inflammation and its immunomodulatory function[D].Master's Thesis.Nanjing:Nanjing Normal University,2016.(in Chinese)
[9] 谢彩虹.嗜酸乳杆菌对抗生素诱导小鼠肠道菌群失调的调整作用[D].硕士学位论文.重庆:第三军医大学,2007. XIE C H.Adjustment effect of Lactobacillus acidophilus on disordered intestinal microbiota of mouse introduced by antibiotics[D].Master's Thesis.Chongqing:The Third Military Medical University,2007.(in Chinese)
[10] 兰桂林,买生,刘晖,等.用嗜酸乳杆菌制剂防治畜禽下痢和提高增重的试验[J].中国兽药杂志,1992,26(1):9-11. LAN G L,MAI S,LIU H,et al.Prevention and control of dysentery and increase of weight gain of livestock and poultry with Lactobacillus acidophilus preparation[J].Chinese Journal of Veterinary Medicine,1992,26(1):9-11.(in Chinese)
[11] LIN M Y,CHANG F J.Antioxidative effect of intestinal bacteria Bifidobacterium longum ATCC 15708 and Lactobacillus acidophilus ATCC 4356[J].Digestive Diseases and Sciences,2000,45(8):1617-1622.  
[12] ZHANG S W,LIU L,SU Y L.Antioxidative activity of lactic acid bacteria in yogurt[J].African Journal of Microbiology Research,2011,5(29):5194-5201.
[13] AMANATIDOU A,BENNIK M H,GORRIS L G,et al.Superoxide dismutase plays an important role in the survival of Lactobacillus sake upon exposure to elevated oxygen[J].Archives of Microbiology,2001,176(1/2):79-88.
[14] LIN M Y,YEN C L.Antioxidative ability of lactic acid bacteria[J].Journal of Agricultural and Food Chemistry,1999,47(4):1460-1466.  
[15] LEE J,HWANG K T,CHUNG M Y,et al.Resistance of Lactobacillus casei KCTC 3260 to reactive oxygen species (ROS):role for a metal ion chelating effect[J].Journal of Food Science,2005,70(8):m388-m391.
[16] 张冰,胡姝敏.干酪乳杆菌对农家干酪抗氧化性及其存活能力的研究[J].中国奶牛,2018(5):50-56. ZHANG B,HU S M.Effect of antioxidant Lactobacillus casei added to cottage cheese on antioxidant activity and survival[J].China Dairy Cattle,2018(5):50-56.(in Chinese)
[17] MIZUTANI A,TSUKAGOSHI N.Molecular role of ascorbate in enhancement of NO production in activated macrophage-like cell line,J774.1[J].Journal of Nutritional Science and Vitaminology,1999,45(4):423-435.  
[18] DI MARZIO L,RUSSO F P,D'AL S,et al.Apoptotic effects of selected strains of lactic acid bacteria on a human T leukemia cell line are associated with bacterial arginine deiminase and/or sphingomyelinase activities[J].Nutrition and Cancer,2001,40(2):185-196.  
[19] MA D L,FORSYTHE P,BIENENSTOCK J.Live Lactobacillus rhamnosus[corrected] is essential for the inhibitory effect on tumor necrosis factor alpha-induced interleukin-8 expression[J].Infection and Immunity,2004,72(9):5308-5314.  
[20] BHATIA S,SHUKLA R,VENKATA MADHU S,et al.Antioxidant status,lipid peroxidation and nitric oxide end products in patients of type 2 diabetes mellitus with nephropathy[J].Clinical Biochemistry,2003,36(7):557-562.  
[21] 黄丽.植物乳杆菌C88的抗氧化作用及安全性评价[D].硕士学位论文.长春:东北师范大学,2012. HUANG L.Antioxidant activity and safety evaluation of Lactobacillus plantarum C88[D].Master's Thesis.Changchun:Northeast Normal University,2012.(in Chinese)
[22] WANG A N,YI X W,YU H F,et al.Free radical scavenging activity of Lactobacillus fermentum in vitro and its antioxidative effect on growing-finishing pigs[J].Journal of Applied Microbiology,2009,107(4):1140-1148.  
[23] HUANG Y P,LONG X,TANG J Y,et al.The attenuation of traumatic brain injury via inhibition of oxidative stress and apoptosis by tanshinone ⅡA[J].Oxidative Medicine and Cellular Longevity,2020,2020:4170156.
[24] LI C,SI J,TAN F,et al.Lactobacillus plantarum KSFY06 prevents inflammatory response and oxidative stress in acute liver injury induced by D-Gal/LPS in mice[J].Drug Design Development and Therapy,2021,15:37-50.
[25] 高世乐,宋金熠,吴菲,等.原花青素对放射性心脏损伤大鼠血清cTn-I、SOD、MDA、GSH-px的影响[J/OL].重庆医科大学学报,2020:1-5.(2020-11-05)[2021-01-13].https://doi.org/10.13406/j.cnki.cyxb.002681.DOI:10.13406/j.cnki.cyxb.002681. GAO S L,SONG J Y,WU F,et al.Effects of proanthocyanidins on serum cTn-I,SOD,MDA,GSH-Px in rats with radiation-induced cardiac injury[J/OL].Journal of Chongqing Medical University,2020:1-5.(2020-11-05)[2021-01-13].https://doi.org/10.13406/j.cnki.cyxb.002681.DOI:10.13406/j.cnki.cyxb.002681.(in Chinese)
[26] WANG B,YANG C T,DIAO Q Y,et al.The influence of mulberry leaf flavonoids and Candida tropicalis on antioxidant function and gastrointestinal development of preweaning calves challenged with Escherichia coli O141:K99[J].Journal of Dairy Science,2018,101(7):6098-6108.  
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