异丹叶大黄素缓解IPEC-J2细胞氧化损伤的效果研究

doi:10.16431/j.cnki.1671-7236.2025.06.038

摘要/Abstract

摘要： 【目的】探究异丹叶大黄素(isorhapontigenin，ISO)对猪小肠上皮细胞(IPEC-J2)氧化损伤的缓解作用，为ISO作为一种抗氧化剂在动物生产中应用提供理论参考。【方法】使用过氧化氢(H₂O₂)处理IPEC-J2细胞构建氧化应激模型，通过细胞存活率筛选出ISO缓解IPEC-J2细胞氧化损伤的最佳浓度；采用相应试剂盒测定细胞中丙二醛(MDA)、活性氧(ROS)、8-羟基脱氧鸟苷(8-OHDG)含量及过氧化氢酶(CAT)、总超氧化物歧化酶(T-SOD)活性和总抗氧化能力(T-AOC)；通过实时荧光定量PCR检测IPEC-J2细胞中抗氧化酶基因(SOD-1、CAT、GSH-Px、HO-1和NQO-1)、炎症相关基因(IL-1β、IL-12和TNF-α)及宿主防御肽基因(pBD1、pBD2和pBD3)的相对表达量。【结果】与H₂O₂组相比，用5、20、40 μmol/L ISO预处理细胞24 h后细胞存活率显著或极显著提高(P＜0.05；P＜0.01)，其中20 μmol/L ISO缓解效果最好。与H₂O₂组相比，ISO预处理可显著或极显著降低IPEC-J2细胞MDA、ROS和8-OHDG水平(P＜0.05；P＜0.01)；极显著提高GSH-Px、SOD1、CAT和NQO-1基因相对表达量及CAT活性(P＜0.01)。与H₂O₂组相比，ISO预处理可降低IL-1β和IL-12基因相对表达量(P＜0.05；P＜0.01)，提高pBD1、pBD2和pBD3基因相对表达量(P＜0.05；P＜0.01)。【结论】ISO可减少细胞中应激产物的生成，提高抗氧化酶基因的表达和抗氧化酶活性，并调节炎症相关基因的表达，从而缓解H₂O₂引起的IPEC-J2细胞氧化损伤。本试验条件下，推荐ISO浓度为20 μmol/L。

关键词: 异丹叶大黄素; 氧化损伤; 抗氧化; IPEC-J2细胞

Abstract: 【Objective】 The objective of this study was to investigate the alleviating effect of isorhapontigenin (ISO) on oxidative damage in porcine intestinal epithelial cells (IPEC-J2),so as to provide a theoretical reference for the application of ISO as an antioxidant in animal production.【Method】 IPEC-J2 cells were stimulated with hydrogen peroxide (H₂O₂) to establish the oxidative stress model,and the optimal working concentration of ISO was determined by the cell viability.The contents of malondialdehyde (MDA),reactive oxygen species (ROS) and 8-hydroxy-deoxyguanosine (8-OHDG),the activities of catalase (CAT),total superoxide dismutase (T-SOD),and total antioxidant capacity (T-AOC) in the cells were determined by the corresponding commercial kit.Real-time quantitative PCR was used to detect the relative expression levels of antioxidase genes (SOD-1, CAT, GSH-Px, HO-1 and NQO-1),inflammatory genes (IL-1β, IL-12 and TNF-α),and host defense peptide genes (pBD1， pBD2 and pBD3) in IPEC-J2 cells.【Result】 Pretreatment of cells with 5,20 and 40 μmol/L ISO for 24 h could alleviate the decrease of cell survival caused by H₂O₂(P＜0.05 or P＜0.01),particularly,the 20 μmol/L ISO group had the best effect among the all groups.ISO pretreatment could reduce the elevation of the levels of MDA,ROS and 8-OHDG in IPEC-J2 cells induced by H₂O₂ (P＜0.05 or P＜0.01) and increase the relative expression levels of GSH-Px, SOD1,CAT and NQO-1 genes and CAT activity (P＜0.01).In addition,compared with H₂O₂ group,ISO pretreatment could decrease the relative expression levels of IL-1β and IL-12 genes,and increase the relative expression levels of pBD1,pBD and pBD3 genes (P＜0.05).【Conclusion】 ISO could reduce the pruduction of stress products in cells,improve the expression of antioxidant enzyme genes and activity of antioxidant enzymes,and regulate the expression of inflammation-related genes,thus alleviating the oxidative damage of IPEC-J2 cells caused by H₂O₂.Under the conditions of this experiment,the recommended ISO concentration was 20 μmol/L.

Key words: isorhapontigenin; oxidative damage; antioxidant; IPEC-J2 cells

中图分类号:

S853.7

朱龙龙, 蔡龙, 刘泽青, 宋妍辰, 吴秋珏, 王晶. 异丹叶大黄素缓解IPEC-J2细胞氧化损伤的效果研究[J]. 中国畜牧兽医, 2025, 52(6): 2865-2873.

ZHU Longlong, CAI Long, LIU Zeqing, SONG Yanchen, WU Qiujue, WANG Jing. Effects of Isorhapontigenin on Alleviating Oxidative Damage of IPEC-J2 Cells[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2865-2873.

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