基于HeLa细胞模型探究植物乳杆菌CY1-2的抗氧化机理  

The Antioxidant Mechanism of Lactiplantibacillus plantarum CY1-2 Based on HeLa Cell Model

作　　者：武文玉 张艳芳 白凤翎 吕欣然 崔方超 檀茜倩 励建荣 李英美 Wu Wenyu;Zhang Yanfang;Bai Fengling;LüXinran;Cui Fangchao;Tan Xiqian;Li Jianrong;Li Yingmei(College of Food Science and Technology,Bohai University,National&Local Joint Engineering Research Center of Storage,Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products,the Fresh Food Storage and Processing Technology Research Institute of Liaoning Provincial Universities,Jinzhou 121013,Liaoning;Dalian Food Jinzhou Co.,Ltd.,Jinzhou 121019,Liaoning;Dalian Polytechnic University,Collaborative Innovation Center of Seafood Deep Processing,Dalian 116034,Liaoning)

机构地区：[1]渤海大学食品科学与工程学院、生鲜农产品贮藏加工及安全控制技术国家地方联合工程研究中心、辽宁省高等学校生鲜食品产业技术研究院,辽宁锦州121013 [2]达莲食品(锦州)有限公司,辽宁锦州121019 [3]大连工业大学、海洋食品精深加工关键技术省部共建协同创新中心,辽宁大连116034

出　　处：《中国食品学报》2024年第8期92-100,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：辽宁省海洋经济发展规划“重点专项”(2021-84);辽宁省“兴辽英才计划”青年拔尖人才项目(XLYC1807133)。

摘　　要：建立植物乳杆菌CY1-2保护偶氮二异丁脒盐酸盐(ABAP)损伤HeLa细胞模型,利用细胞增殖试验、细胞形态学观察及免疫印迹试验探究植物乳杆菌CY1-2的细胞保护机制。结果表明,当菌液浓度为108 CFU/mL的植物乳杆菌CY1-2保护细胞3 h后,再用浓度为110 mmol/L的ABAP损伤细胞3 h。在此模型条件下,植物乳杆菌CY1-2菌体及无细胞提取物均能有效提高HeLa细胞的存活能力,细胞存活率分别提高9.41%和25.71%。通过显微镜观察发现,与损伤组相比,经植物乳杆菌CY1-2无细胞提取物保护的HeLa细胞大而饱满,边缘清晰且伸展性好。免疫印迹试验表明,与损伤组相比,植物乳杆菌CY1-2无细胞提取物能够正向调控HeLa细胞的kelch样ECH关联蛋白1(Keap1)、核因子E2-相关因子(Nrf2)和血红素加氧酶-1(HO-1)蛋白表达量,分别提高了18%,47%和58%。植物乳杆菌CY1-2保护ABAP氧化损伤HeLa细胞的机制是激活细胞Keap1-Nrf2-ARE信号通路,提高其抗氧化能力。本研究结果为开发乳酸菌抗氧化剂提供一定的理论依据。In the study,a model of HeLa cells protected by Lactiplantibacillus plantarum CY1-2 against 2,2'-azobis[2-methylpropiona midine]dihydrochloride(ABAP)damage was established.The antioxidant mechanism of L.plantarum CY1-2 was investigated using cell proliferation test,morphological observation and western blot analysis.The results showed that the cell protective model conditions were as follows:108 CFU/mL of L.plantarum CY1-2 protected cells for 3 h,and then 110 mmol/L of ABAP damaged cells for 3 h.Under this model condition,both the cell and the cell-free extract of L.plantarum CY1-2 could effectively improve the viability of HeLa cells,and the cell survival rate increased by 9.41%and 25.71%,respectively.Meanwhile,microscopic observation showed that compared with the injured group,HeLa cells protected by the cell-free extract of L.plantarum CY1-2 were large and plump,with clear edges and good extensiability.Western blot analysis indicated that,compared with the injured group,the cell-free extract of L.plantarum CY1-2 could positively regulate the expression of kelch-like ECH-associated protein 1(Keap1),nuclear factor erythroid 2-related factor(Nrf2)and heme oxygenase-1(HO-1)proteins in HeLa cells,resulting in increasing by 18%,47%and 58%respectively.L.plantarum CY1-2 protects HeLa cells from oxidative damage by activating Keap1-Nrf2-ARE signaling pathway to improve its antioxidant capacity.This study was expected to provide a theoretical basis for the development of lactic acid bacteria antioxidants.

关 键 词：植物乳杆菌 氧化损伤 HeLa细胞模型 抗氧化 

分 类 号：TS201.3[轻工技术与工程—食品科学]