超氧化物歧化酶模拟物对氧化应激IPEC-J2细胞抗氧化性能的影响

doi:10.16431/j.cnki.1671-7236.2021.05.007

摘要/Abstract

摘要： 试验旨在探究在H₂O₂诱导的氧化应激状态下，超氧化物歧化酶模拟物（SODm）对仔猪空肠上皮细胞系（IPEC-J2）的保护作用。利用MTT法筛选出构建氧化应激模型H₂O₂的适宜浓度；将IPEC-J2细胞分别用0、0.05、0.5、5、50、500、2 000 U/mL SODm进行培养，利用MTT法分别在2、4、8、12 h时测定各组细胞存活率，筛选出SODm作用的适宜浓度和时间；根据构建的氧化应激模型和SODm适宜浓度和时间，将IPEC-J2细胞随机分为空白组、模型组、SODm处理组（SODm_0.5、SODm₅、SODm₅₀）和超氧化物歧化酶（SOD）处理组（SOD_0.5、SOD₅、SOD₅₀），分别测定各组细胞存活率、活性氧（ROS）含量及细胞内SOD、谷胱甘肽过氧化物酶（GSH-Px）活性及丙二醛（MDA）含量和总抗氧化能力（T-AOC）。结果表明：①H₂O₂构建细胞氧化应激模型的适宜浓度是1.0 mmol/mL。②当不同浓度SODm分别作用4、8、12 h时，0.5~50 U/mL SODm组细胞存活率显著高于空白组（P<0.05）；且8 h时存活率最高，在12 h时处于下降趋势。③除空白组外，SODm₅和SOD₅预处理的IPEC-J2存活率显著高于其他组（P<0.05）；且SODm和SOD组中细胞ROS含量显著低于模型组（P<0.05）；模型组与空白组相比，均显著降低了SOD、GSH-Px活性和T-AOC水平，提高了MDA含量（P<0.05）；与模型组相比，所有SODm和SOD处理组均显著提高了SOD、GSH-Px活性和T-AOC水平，降低了MDA含量（P<0.05），同时SODm₅₀组T-AOC水平显著低于SOD₅₀组（P<0.05）。综上，SODm对H₂O₂诱导氧化损伤状态下IPEC-J2具有保护作用。

关键词: 超氧化物歧化酶模拟物(SODm); 氧化应激; IPEC-J2细胞

Abstract: The objective of this study was to investigate the protective effect of superoxide dismutase mimetic (SODm) on intestinal epithelial cell (IPEC-J2) under H₂O₂-induced oxidative stress.Using MTT method to screen out the proper concentration of H₂O₂ applied in structuring oxidative stress model.IPEC-J2 were cultured with 0,0.05,0.5,5,50,500 and 2 000 U/mL SODm,respectively.The survival of each group was detected by MTT in 2,4,8 and 12 h to determine the proper range and duration of a SODm dose.Based on the model of oxidative stress and the proper range and duration of SODm,IPEC-J2 were randomly grouped as the control group,the model group with H₂O₂,the disposed group with SODm (SODm_0.5,SODm₅,SODm₅₀),the disposed group with SOD (SOD_0.5,SOD₅,SOD₅₀),then cell viability,ROS levels and SOD,GSH-Px,MDA,T-AOC levels in cells were measured in each group.The results were showed as follows:①1.0 mmol/mL H₂O₂ was the proper concentration in structuring oxidative stress model.②The survival ratios of the disposed group with SODm of 0.5~50 U/mL in 4,8 and 12 h was significantly higher than that of the control groups (P<0.05).The survival ratio was highest at 8 h and decreased at 12 h.③The IPEC-J2 survival rate of the SODm₅ and SOD₅ groups were significantly higher than that in other groups (P<0.05) except for the blank group,and the ROS contents in the SODm and SOD groups were significantly lower than that in the model group (P<0.05).The activities of SOD,GSH-Px and T-AOC were significantly decreased,and MDA content were significantly increased in model group compared with the control group (P<0.05).The treatments of SODm and SOD increased the activities of SOD,GSH-Px and T-AOC,and significantly decreased MDA content compared with the model group(P<0.05),The T-AOC level of SOD₅₀ group was higher than that of SODm₅₀ group (P<0.05).In conclusion,SODm had protective effect on IPEC-J2 cells under H₂O₂-induced oxidative stress.

Key words: superoxide dismutase mimetic(SODm); oxidative stress; IPEC-J2 cells

中图分类号:

王思博, 崔红霞, 杨季, 许丽. 超氧化物歧化酶模拟物对氧化应激IPEC-J2细胞抗氧化性能的影响[J]. 中国畜牧兽医, 2021, 48(5): 1566-1573.

WANG Sibo, CUI Hongxia, YANG Ji, XU Li. Effect of Superoxide Dismutase Mimetic on Antioxidant Capacity of Oxidatively Damaged IPEC-J2 Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1566-1573.

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