摘要
目的:通过脂多糖诱导巨噬细胞RAW264.7建立炎症和氧化应激模型,探讨人参总皂苷的抗炎和抗氧化效果以及对脂多糖诱导的巨噬细胞RAW264.7自噬水平的影响。方法:培养RAW264.7细胞,给予脂多糖刺激并加入不同浓度人参总皂苷,使用硝酸还原酶法检测细胞内一氧化氮水平;ELISA法检测小鼠肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的分泌;DCFH-DA荧光探针法检测人参总皂苷处理后活性氧的变化;超氧化物阴离子荧光探针(dihydroethidium,DHE)检测细胞内超氧化物阴离子水平;吖啶橙染色法检测自噬体的形成;Western Blot法检测COX-2、NF-кB、Nrf2、HO-1、GCLC、GCLM、Beclin1、ATG7和LC3的蛋白表达。结果:与脂多糖模型组相比,随着TGS的浓度增加,细胞一氧化氮水平、TNF-α的分泌降低,细胞内活性氧和超氧阴离子的含量降低,巨噬细胞RAW264.7酸性自噬体的数量增加;Nrf2、HO-1、GCLC、GCLM、Beclin1、ATG7及LC3的蛋白表达提高,COX-2、NF-кB的蛋白表达降低。结论:在RAW264.7巨噬细胞中,人参总皂苷可通过NF-кB、Nrf2/ARE通路以及提高自噬水平来降低脂多糖引起的炎症反应和氧化损伤。
OBJECTIVE To explore the anti-inflammatory and antioxidant effects of total ginsenoside(TGS)and its effects on autophagy level of lipopolysaccharide(LPS)-induced macrophages RAW264.7 based on LPS-induced inflammation and oxidative stress models of macrophages RAW264.7.METHODS RAW264.7 cells were cultured,and then stimulated by LPS,and different concentrations of TGS were added to the cells.Nitrate reductase method was used to detect intracellular nitric oxide(NO)level;ELISA method was used to detect TNF-αsecretion;DCFH-DA fluorescent probe method was used to detect TGS changes of ROS after treatment;DHE fluorescent probe was used to detect the level of superoxide anion in the cells;acridine orange staining method was used to detect autophagosome formation;Western Blot method was used to detect the protein expression of NF-кB,Nrf2,HO-1,GCLC,GCLM,LC3,ATG7,Beclin1,and COX-2.RESULTS Compared with the LPS model group,with the increase of the concentration of TGS,the level of cellular NO and the secretion of TNF-αgradually decreased,and the contents of intracellular ROS and superoxide anion decreased.TGS increased the acidic autophagy of macrophage RAW264.7,as well as increased the protein expression of Nrf2,HO-1,GCLC,GCLM,Beclin1,ATG7 and LC3,and decreased the protein expression of COX-2 and NF-кB.CONCLUSION In RAW264.7 macrophages,total ginsenoside can reduce the inflammatory response and oxidative damage caused by LPS through NF-кB and Nrf2/ARE pathways,and increase the level of autophagy.
作者
任李梅
杨松
卢姝言
赵悦名
李文婧
张巍
王佳雯
REN Li-mei;YANG Song;LU Shu-yan;ZHAO Yue-ming;LI Wen-jing;ZHANG Wei;WANG Jia-wen(Jilin Ginseng Academy,Changchun University of Chinese Medicine,Jilin Changchun 130000,China;Research Office,Changchun University of Chinese Medicine,Jilin Changchun 130000,China)
出处
《中国医院药学杂志》
CAS
北大核心
2022年第9期896-901,共6页
Chinese Journal of Hospital Pharmacy
基金
吉林省科技发展计划项目(编号:20210204146YY)。
