摘要
目的:探讨黄芪甲苷对慢性阻塞性肺疾病(COPD)的影响及通过Nrf2/HO-1轴对氧化应激的作用。方法:CCK-8筛选黄芪甲苷对A549细胞作用的安全范围,利用香烟提取物构建COPD细胞模型。将细胞分为正常对照组、模型组及黄芪甲苷25、50、100μg/mL组,CCK-8检测细胞活力,流式细胞术检测细胞内ROS水平及细胞凋亡情况,生化法检测细胞上清中SOD、MDA水平。PCR和Western Blot检测细胞中Nrf2、HO-1 mRNA及蛋白表达。为了进一步验证黄芪甲苷对Nrf2/HO-1的影响,用Nrf2抑制剂ML385处理细胞,CCK-8检测细胞活力,流式细胞术检测细胞内ROS水平。结果:黄芪甲苷浓度安全范围为25~100μg/mL。黄芪甲苷干预后能提高细胞活力、SOD水平和Nrf2、HO-1 mRNA及蛋白表达,降低ROS、MDA水平和细胞凋亡率(P<0.01),其作用呈浓度依赖性,其中100μg/mL效果最好。ML385可逆转黄芪甲苷的作用(P<0.01)。结论:黄芪甲苷可通过降低ROS水平,调节Nrf2/HO-1轴来抑制氧化应激,从而治疗COPD。
Objective:To explore the effect of astragalosideⅣon chronic obstructive pulmonary disease(COPD)and the effect of on oxidative stress through Nrf2/HO-1 axis.Methods:The safe range of astragalosideⅣon A549 cells was screened by CCK-8,and the COPD cell model was constructed by cigarette extract.The cells were divided into normal control group,model group and astragalosideⅣ(25,50,100μg/mL)groups.CCK-8 was used to detect cell activity,intracellular ROS level and apoptosis of cells were detected by flow cytometry,and the SOD and MDA levels in the supernatant were detected by biochemical method.The mRNA and protein expressions of Nrf2 and HO-1 were detected by PCR and Western Blot.To further verify the effect of astragalosideⅣon Nrf2/HO-1,cells were treated with Nrf2 inhibitor ML385,cell activity was detected by CCK-8,and intracellular ROS level was detected by flow cytometry.Results:The safe concentration range of astragalosideⅣwas 25 to 100μg/mL.AstragalosideⅣcould increase the cell viability,SOD level,Nrf2,HO-1 mRNA and proteins expressions,decrease ROS,MDA levels and apoptosis rate(P<0.01)in a content-dependent manner,and 100μg/mL had the best effect.ML385 could reverse the effect of astragalosideⅣ(P<0.01).Conclusion:AstragalosideⅣcan treat COPD by reducing ROS levels and regulating Nrf2/HO-1 axis to inhibit oxidative stress.
作者
沈三英
谈小琴
SHEN San-ying;TAN Xiao-qin(Wuhan Fourth Hospital,Wuhan 430033,China)
出处
《中药材》
CAS
北大核心
2022年第12期2975-2981,共7页
Journal of Chinese Medicinal Materials
基金
武汉市卫生计生委员会临床医学科研基金资助项目(WZ15B08,WZ17B09)
湖北省科技厅科技条件资源开发项目(2015BCE099)
