摘要
本研究通过脂多糖(LPS)诱导大鼠腹腔巨噬细胞(RAW264.7)建立氧化应激模型,探讨替唑尼特(TIZ)的抗氧化和抗炎效果。培养RAW264.7细胞,给予LPS(1μg/m L)刺激并用TIZ(25、50、75、100μmol/L)作用。MTT法检测细胞存活率;DCFH-DA荧光探针法检测TIZ对ROS生成的影响;同时,检测MDA的生成量以评估TIZ对脂质氧化的影响;检测GSH的含量以及CAT、SOD、GSH-Px酶活性以评估TIZ对细胞抗氧化系统的影响;检测IL-6的分泌和COX-2的表达量以评估TIZ对炎症细胞因子的作用。结果显示LPS刺激后细胞表现出显著性的氧化应激效应,而给予TIZ处理后,ROS和MDA生成量呈剂量相关性的显著下降;细胞IL-6的分泌和COX-2的表达量也随TIZ处理而显著下降,而GSH含量以及CAT、SOD、GSH-Px活性在TIZ处理后却呈剂量相关性的显著性上升。因此,TIZ可抑制LPS诱导的RAW264.7细胞氧化应激和炎症反应,这为TIZ的作用机制研究以及应用奠定了基础。
The oxidative stress model was developed to investigate the antioxidant effect and anti-inflammatory of TIZ through induction of RAW264.7 cells with LPS. The RAW264.7 cells were cultured and treated with LPS (1 μg/mL) and TIZ at 25, 50, 75, 100 μmol/L for 12 h. The cell viability was measured by MTT assay. DCFH-DA fluorescence probe was used to detect the effect of TIZ on the generation of reactive oxygen species. The MDA production was measured to evaluate the effect of TIZ on lipid oxidation. Additionally, the effect of TIZ on the antioxidant activity of the treated RAW264.7 cells was assessed for the content of GSH and activity of CAT, SOD and GSH-Px, and inflammatory cytokines for the expressions of IL-6 and COX-2. The results showed that LPS stimulation induced significant oxidative stress effect while TIZ treatment caused significant decrease of the production of reactive oxygen species and MDA production. The secretion of IL-6 and the expression of COX-2 also significantly decreased in a dose-dependent manner. In contrast, the content of GSH and activity of CAT, SOD and GSH-Px increased significantly in a dose-dependent manner after TIZ treatment. Thus, TIZ treatment inhibited oxidative stress and inflammation on RAW264.7 cells but LPS induced an increase. These results laid a solid foundation for further study on the mechanism and application of TIZ.
作者
首姣琴
程晓蕾
王霄旸
薛飞群
王米
刘迎春
费陈忠
张丽芳
张可煜
李娟
SHOU Jiao-qin;CHENG Xiao-lei;WANG Xiao-yang;XUE Fei-qun;WANG Mi;LIU Ying-chun;FEI Chen-zhong;ZHANG Li-fang;ZHANG Ke-yu;LI Juan(College of Chemistry,Xiangtan University,Xiangtan 411100;Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics,Ministry of Agriculture and Rural Affairs,Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)
出处
《中国动物传染病学报》
CAS
北大核心
2019年第2期71-77,共7页
Chinese Journal of Animal Infectious Diseases
基金
国家科技支撑计划项目(2015BAD11B00)
国家自然科学基金(31872516)
