摘要
目的:研究黄芪总黄酮对骨关节炎软骨细胞氧化应激和炎症因子分泌的影响及机制。方法:分离培养骨关节炎软骨细胞,分别用0、5、10、20mg/L的黄芪总黄酮处理细胞,72h后采用MTT方法检测细胞的增殖能力,收集培养上清,ELISA法检测白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)含量,WST-8法检测超氧化物歧化酶(SOD)活性,ELISA法检测谷胱甘肽过氧化物酶(GSH-Px)活性,Westernblot法检测细胞中NF-κBp65亚型(NF-κBp65)、基质金属蛋白酶-13(MMP-13)蛋白表达水平。结果:5、10、20mg/L黄芪总黄酮处理后的骨关节炎软骨细胞的增殖能力升高,细胞培养上清中IL-6、IL-1β含量下降,SOD和GSH-Px活性升高,细胞中NF-κBp65、MMP-13蛋白表达水平降低(P<0.05)。结论:黄芪总黄酮可抑制骨关节炎软骨细胞氧化应激和炎症因子分泌,作用机制可能与下调NF-κB信号通路有关。
Aim :To study the effects and mechanism of total flavonoids of Astragalus on oxidative stress and inflammatory cytokine secretion in chondrocytes of osteoarthritis. Methods :After isolation and culture of osteoarthritis chondrocytes, the cells were treated with 0, 5, 10 and 20 mg/L total flavonoids of Astragalus,respectively.MTT assay was used to detect the proliferation after 72 hours. ELISA method was used to detect the content of IL-6 and IL-1β in the supernatant of cell culture medium. The activity of SOD in supernatant was detected by WST-8. The activity of GSH-Px in supernatant was detected by ELISA. Western blot assay was used to detect the levels of NF-κBp65 and MMP-13 proteins in the cells. Results :The proliferative capacity of articular chondrocytes increased after treatment with 5, 10 and 20 mg/L total flavonoids of Astragalus, the contents of IL-6 and IL-1β in the supernatant of cell culture medium decreased, the activity of SOD and GSH-Px increased,and the expression levels of NF-κBp65 and MMP-13 decreased( P <0.05). Conclusion :Total flavonoids of Astragalus could inhibit oxidative stress and inflammatory cytokine secretion in chondrocytes of osteoarthritis,and the mechanism may be related to down-regulation of NF-κB signal pathway.
作者
任伟亮
焦永伟
张健
王响
齐立卿
杜双庆
REN Weiliang;JIAO Yongwei;ZHANG Jian;WANG Xiang;QI Liqing;DU Shuangqing(Third Ward of Department of Orthopedics,Hebei Provincial Hospital of Traditional Chinese Medicine, Shijiazhuang 050011)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2019年第4期603-606,共4页
Journal of Zhengzhou University(Medical Sciences)
基金
河北省中医药管理局科研计划项目(2016166)
关键词
黄芪总黄酮
骨关节炎软骨细胞
NF-ΚB
炎症
氧化应激
total flavonoids of Astragalus
chondrocyte of osteoarthritis
NF-κB
inflammation
oxidative stress
