摘要
为探讨邻苯二甲酸二(2-乙基己)酯(DEHP)暴露对MAC-T细胞的毒性作用及橙皮苷(HDN)的干预效果,试验分为4组,对照组、DEHP暴露组、DEHP+N-乙酰半胱氨酸(NAC)干预组、DEHP+HDN干预组,利用RT-qPCR分析抗氧化相关基因(NQO1、HMOX1、GPX1、GPX4)和炎症相关基因(IL-6、IL-1β、TNF-α、CCL2)的mRNA表达量;应用试剂盒检测氧化应激指标(MDA含量、CAT和T-SOD活性、ROS含量);应用ELISA法检测炎性因子(IL-6、IL-1β、TNF-α)分泌量;采用蛋白质印迹法(Westernblot)检测IL-6、IL-1β、TNF-α蛋白相对表达量。结果表明:与对照组相比,DEHP组抗氧化相关基因mRNA表达量显著下调、MDA含量显著升高、CAT和T-SOD活性显著降低、ROS含量显著升高、炎症相关基因mRNA及蛋白表达量显著升高;抗氧化剂NAC可显著抑制DEHP诱导的炎症反应;HDN干预后,抗氧化相关基因mRNA表达量显著升高、MDA含量显著降低、CAT和T-SOD活性升高、ROS含量显著降低、炎症相关基因mRNA及蛋白表达量显著下调。DEHP暴露可引起MAC-T细胞氧化应激和炎症损伤,HDN可通过增强抗氧化能力及抑制炎症反应发挥保护作用。
This study aimed to investigate the toxic effects of di(2-ethylhexyl)phthalate(DEHP)on MAC-T cells and the interventional effect of hesperidin(HDN).Four groups were established as follows:control group,DEHP exposure group,DEHP+N-Acetylcysteine(NAC)intervention group,and DEHP+HDN intervention group.RT-qPCR was employed to examine the mRNA expression of antioxidant-related genes(NQO1,HMOX1,GPX1 and GPX4)and inflammation-related genes(IL-6,IL-1β,TNF-αand CCL2).Com-mercial kits were used to detect oxidative stress indicators(MDA content,CAT and T-SOD activities,ROS content).ELISA was utilized to determine the secretion of inflammatory cytokines(IL-6,IL-1βand TNF-α),and Western blot was used to detect the relative protein expression of these cytokines.The results showed that compared with the control group,the DEHP group showed significantly downregulated mRNA expression of antioxidant-related genes,significantly increased MDA and ROS contents,significantly decreased CAT and T-SOD activities,and significantly elevated mRNA and protein expression of inflammation-related cytokines.The antioxidant NAC significantly inhibited DEHP-induced inflammatory responses.After HDN intervention,the mRNA expression of antioxidantrelated genes was significantly increased,the MDA content was significantly decreased,the activities of CAT and T-SOD were increased,the ROS levels were significantly decreased,and both the mRNA and protein levels of inflammation-related cytokines were significantly downregulated.DEHP exposure induces oxidative stress and inflammatory damage in MAC-T cells,whereas HDN exerts a protective effect by enhancing antioxidant capacity and inhibiting inflammatory responses.
作者
邵雯琪
陈芳芳
王锦浩
刘克祥
赵树臣
赵立佳
SHAO Wenqi;CHEN Fangfang;WANG Jinhao;LIU Kexiang;ZHAO Shuchen;ZHAO Lijia(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;Key Laboratory of the Provincial Education Department of Heilongjiang for Common Animal Disease Prevention and Treatment,Northeast Agricultural University,Harbin 150030,China)
出处
《东北农业大学学报》
北大核心
2025年第6期135-145,共11页
Journal of Northeast Agricultural University
基金
黑龙江省自然科学基金优秀青年基金项目(YQ2022C018)。
