细胞代次对t-BHP诱导Caco-2细胞构建氧化应激模型的影响  

Effect of Cell Generations on t-BHP-induced Oxidative Stress Model of Caco-2 Cells

作　　者：倪俊 袁彩莲 佘容 赵成法 李丽娟 杨旭 杨晓燕 NI Jun;YUAN Cailian;SHE Rong;ZHAO Chengfa;LI Lijuan;YANG Xu;YANG Xiaoyan(Institute of Natural Antioxidants and Anti-inflammation,Dali University,Dali 671003,China;Institute of Eastern-himalaya,Dali University,Dali 671003,China;School of Public Health,Dali University,Dali 671003,China;The Provincial Innovation Team of Biodiversity Conservation and Utility of the Three Parallel Rivers Region from Yunnan Province,Dali 671003,China;Collaborative Innovation Center for Biodiversity and Conservation in the Three Parallel Rivers Region of China,Dali 671003,China)

机构地区：[1]大理大学天然抗氧化剂和抗氧化炎症研究院,云南大理671003 [2]大理大学东喜玛拉雅研究院,云南大理671003 [3]大理大学公共卫生学院,云南大理671003 [4]三江并流区域生物多样性保护与利用云南省创新团队,云南大理671003 [5]中国三江并流区域生物多样性协同创新中心,云南大理671003

出　　处：《食品工业科技》2023年第5期60-66,共7页Science and Technology of Food Industry

基　　金：云南省科技厅科技计划项目,“氧化炎症诊断测量仪器的研制:血液ROS、MDA和尿8-OHdG的检测”(202101BA070001-115)。

摘　　要：目的:探究细胞代次对叔丁基过氧化氢(t-BHP)诱导构建人克隆结肠腺癌细胞(Caco-2)氧化应激模型的影响。方法:以不同浓度t-BHP(1、2和3 mmol/L)处理第F15代Caco-2细胞1、2、3、4、5和6 h后测定不同处理下细胞丙二醛(MDA)和8-羟基脱氧鸟苷(8-OHdG)的含量,以确定构建Caco-2细胞氧化应激模型的基础条件。再以8-OHdG为分子标志物,探究细胞代次(F14、F15、F18、F19和F20)对Caco-2细胞氧化应激模型构建的影响。结果:采用1、2和3 mmol/L t-BHP处理F15代Caco-2细胞3~6 h均可稳定构建氧化应激模型。当以2.0 mmol/L t-BHP处理不同代次Caco-2细胞5 h后,F14、F15和F18代细胞的8-OHdG浓度均显著高于对照(P<0.05),而F19和F20代细胞的8-OHdG浓度与对照无显著差异。结论:Caco-2细胞代次会影响其氧化应激模型的构建,F19代之前的Caco-2细胞采用2 mmol/L t-BHP处理细胞5 h可稳定构建氧化应激模型。Objective:To investigate the effects of cell passage number on the establishment of an oxidative stress model in human colon adenocarcinoma cells(Caco-2)caused by tert-butyl hydroperoxide(t-BHP).Methods:First,by applying various t-BHP concentrations(1,2 and 3 mmol/L)to the F15 generation of Caco-2 cells for 1,2,3,4,5 and 6 hours,the content of malondialdehyde(MDA)and 8-hydroxydeoxyguanosine(8-OHdG)in different treatment groups were measured to determine the basic conditions for constructing the oxidative stress model of Caco-2 cells.Then,based on the fundamental parameters,the effect of cell passage number(F14,F15,F18,F19 and F20)on the development of oxidative stress model was investigated using the content of 8-OHdG as an index.Results:When exposed to 1,2 and 3 mmol/L of tBHP for 3 to 6 hours,the oxidative stress model of Caco-2 cells from the F15 generation could be established steadily.The content of 8-OHdG in Caco-2 cells of the F14,F15 and F18 generations was significantly higher than that in control group(which had not been treated with t-BHP)after being exposed to 2.0 mmol/L t-BHP for 5 hours(P<0.05),whereas there was no significant difference between the F19 and F20 generations and the control group.Conclusion:The development of the oxidative stress model of Caco-2 cells will be strongly impacted by cell passage number.By subjecting Caco-2 cells prior to F19 to 2 mmol/L t-BHP for 5 hours,the oxidative stress model may be successfully constructed.

关 键 词：CACO-2细胞 叔丁基过氧化氢 氧化应激 细胞模型 细胞代次 8-羟基脱氧鸟苷 

分 类 号：Q256[生物学—细胞生物学]