食品科学 ›› 2020, Vol. 41 ›› Issue (2): 173-180.doi: 10.7506/spkx1002-6630-20181120-225

• 生物工程 • 上一篇    下一篇

具抗氧化功能益生菌菌株筛选及其对丙烯酰胺诱导肠上皮细胞氧化损伤的保护作用

李桐,吴思琪,曹鑫,宋静颐,张红星,谢远红,金君华   

  1. (食品质量与安全北京实验室,农产品有害微生物及农残安全检测与控制北京市重点实验室,北京农学院食品科学与工程学院,北京 102206)
  • 出版日期:2020-01-25 发布日期:2020-01-19
  • 基金资助:
    北京市属高校青年拔尖人才培育计划项目(CIT&TCD201804039)

Screening for Antioxidant Probiotics and Their Protective Effect on Oxidative Damage Induced by Acrylamide in Intestinal Epithelial Cells

LI Tong, WU Siqi, CAO Xin, SONG Jingyi, ZHANG Hongxing, XIE Yuanhong, JIN Junhua   

  1. (Beijing Laboratory of Food Quality and Safety, Beijing Key Laboratory of Agricultural Product Detection and Control of Spoilage Organisms and Pesticide Residue, College of Food Science and Engineering, Beijing University of Agriculture, Beijing 102206, China)
  • Online:2020-01-25 Published:2020-01-19

摘要: 目的:以1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率为指标,筛选抗氧化能力强的乳杆菌,并研究其对模拟胃肠道环境的耐受性及对丙烯酰胺所致肠上皮细胞氧化损伤的保护作用。方法:以DPPH自由基清除率为指标,筛选得到清除率高的2 株乳杆菌GBE17和GBE29,并进行16S rDNA测序鉴定。构建丙烯酰胺诱导的Caco-2氧化损伤模型,通过形态学观察和测定细胞培养上清液、细胞裂解液中的抗氧化物相关物质及酶活,评价其不同处理方式对Caco-2细胞氧化损伤的保护作用,并分别测定其在pH 3.0、2.5及胆盐质量分数为0.05%、0.1%环境下处理1~4 h的活菌数变化。结果:植物乳杆菌GBE17和唾液乳杆菌GBE29发酵上清液的DPPH自由基清除率分别为89.44%、79.24%。GBE17的治疗组与干预组和GBE29的干预组均可降低乳酸脱氢酶的释放,提高细胞内外超氧化物歧化酶、过氧化氢酶活性。2 株乳杆菌在pH 3.0和胆盐质量分数0.1%的环境下处理4 h,存活数均高于107 CFU/mL。结论:植物乳杆菌GBE17和唾液乳杆菌GBE29通过提高细胞内抗氧化酶系的活性,有效地降低丙烯酰胺诱导肠上皮细胞的氧化损伤,且具有较好的胁迫耐受能力。

关键词: 乳酸菌, 氧化损伤, 丙烯酰胺, 耐受性, Caco-2细胞

Abstract: Objectives: To screen for lactobacilli with high antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals in vitro and to evaluate their tolerance to simulated gastrointestinal environment and their protective effect against oxidative injury induced by acrylamide in intestinal epithelial cells. Methods: Lactobacillus GBE17 and GBE29, which were found to have strong antioxidant potential, were identified by were identified by 16S rDNA sequencing. The protective effect of the viable cells and cell-free culture supernatant of either strain on acrylamide-induced oxidative injury in Caco-2 cells was assessed by morphological observation and measurement of antioxidant compounds contents and enzyme activities in the cell culture supernatants and lysates. Changes in the viable cell count after 1–4 h of exposure to different pHs (3.0 and 2.5) and different bile salt concentrations (0.05% and 0.1%) were monitored. Results: The scavenging percentages of DPPH radical by the culture supernatants of L. plantarum GBE17 and L. salivarius GBE29 were 89.44% and 79.24%, respectively. Both treatment and prevention with GBE17 as well as prevention with GBE29 could increase cell viability by decreasing lactate dehydrogenase activity and increasing the activities of superoxide dismutase and catalase in Caco-2 cells. For both strains, the number of cells surviving pH 3.0 and 0.1% bile salt environments for 4 h was higher than 7.0 (lg(CFU/mL)). Conclusion: L. plantarum GBE17 and L. salivarius GBE29 have stress tolerance and show a protective effect against acrylamide-induced oxidative damage in Caco-2 cells by increasing intracellular antioxidant enzymes activities.

Key words: Lactobacillus, oxidative damage, acrylamide, resistance, Caco-2 cells

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