摘要:
目的 采用脂多糖(lipopolysaccharide,LPS)诱导星形胶质细胞C6的氧化应激模型,以研究酸枣仁皂苷A(Jujuboside A,JuA)的抗氧化活性.方法 体外培养大鼠星形胶质细胞C6.采用MTT法检测各组细胞的存活率,采用Western blot法检测星形胶质细胞标记蛋白(glial fibrillary acidic protein,GFAP)的活化情况,采用Griess还原法测定细胞上清液中一氧化氮(nitric oxide,NO)水平,采用流式细胞术检测各组细胞中活性氧(reactive oxygen species,ROS)水平,采用ELISA测定各组细胞内丙二醛(malondialdehyde,MDA)和谷胱甘肽(L-glutathione,GSH)水平以及超氧化物歧化酶(superoxide dismutase,SOD)活性.结果 0.125~50 μmol/L JuA对C6细胞存活率无明显影响(P>0.05);1 μg/mL LPS可以明显升高C6细胞GFAP蛋白表达水平,显著升高细胞上清液中NO和细胞内ROS、MDA水平,降低细胞内GSH水平和SOD活性,差异均有统计学意义(P<0.05).与LPS组比较,12.5、25、50μmol/L JuA可以明显抑制LPS诱导的GFAP表达,降低细胞上清液中NO水平,降低细胞内ROS、MDA水平,升高细胞内GSH水平和SOD活性,差异均有统计学意义(P<0.05).结论 JuA对LPS诱导的星形胶质细胞氧化损伤具有一定的保护作用.%Objective To investigate the antioxidant activity of jujuboside A (JuA) by establishing a model of lipopolysaccharide (LPS)-induced oxidative stress in C6 astrocytes.Methods Rat C6 astrocytes were cultured in vitro.MTT assay was used to measure the viability of C6 astrocytes;Western blot was used to measure the activation of glial fibrillary acidic protein (GFAP);the Griess method was used to measure the level of nitric oxide (NO) in supernatant;flow cytometry was used to measure the level of reactive oxygen species (ROS) in C6 astrocytes;ELISA was used to measure the levels of malondialdehyde (MDA) and L-glutathione (GSH) and the activity of superoxide dismutase (SOD) in C6 astrocytes.Results JuA at a concentration of 0.125-50 μmol/L had no significant influence on the viability of C6 astrocytes (P>0.05).1 μg/mL LPS significantly increased the protein expression of GFAP in C6 astrocytes,the level of NO in supernatant,and the levels of ROS and MDA in C6 astrocytes and significantly reduced the level of GSH and the activity of SOD in C6 astrocytes (all P<0.05).Compared with LPS,12.5,25,and 50 μmol/L JuA significantly inhibited the protein expression of GFAP induced by LPS,reduced the level of NO in supernatant and the levels of ROS and MDA in C6 astrocytes,and increased the level of GSH and the activity of SOD in C6 astrocytes (all P<0.05).Conclusion JuA has a certain protective effect against LPS-induced oxidative damage in astrocytes.