羟基酪醇对LPS刺激的THP-1细胞氧化应激的抑制作用

Suppressive effects of hydroxytyrosol on oxidative stress of THP-1 cells stimulated by LPS

目的:主要研究羟基酪醇对脂多糖(LPS)刺激的THP-1细胞引起的氧化应激以及NF-κB活化的抑制作用。方法:以2′,7′-二氢二氯荧光素二乙酸酯(DCFH-DA)为荧光探针检测细胞内活性氧(Reactive oxygen species,ROS)水平;以荧光法检测细胞内的谷胱甘肽(Glutathione,GSH)的水平;以测定细胞内的硝酸盐含量来检测细胞内的NO水平;以蛋白印记法来检测NF-κB的表达水平。结果:羟基酪醇能够剂量依赖性地降低细胞内由LPS刺激的THP-1细胞引起的ROS和NO水平的升高;50μmol/L和100μmol/L的羟基酪醇能增加细胞内的GSH水平;并且能够抑制由LPS引起的NF-κB的活化。抗氧化性的N-乙酰半胱氨酸(NAC)对NO、ROS、GSH以及NF-κB的影响与羟基酪醇一致。结论:羟基酪醇在THP-1细胞具有很强的抗氧化性,能抑制LPS诱导的氧化应激以及NF-κB的活化。

Objective：To investigate whether hydroxytymsol （HT） may ameliorate oxidative stress and nuclear factor kappa B （NF-κB） activation in the lipopolysaccharide （LPS）-stimulated THP-1 cell line. Methods：The intracellular formation of reactive oxygen species （ROS） was detected by 2,7-dichlomfluorescein diacetate （DCFH-DA） as a fluorescent probe. Intracellular glutathione （GSH） level was estimated by fluorometric methods; Nitric oxide （NO） production was measured as nitrite （a stable metabolite of NO） concentrations. Western blot analysis was used to detect the expressin level of NF-κB.Results：The results showed that treatment of THP-1 cells with HT significandy reduced LPS- stimulated NO production and ROS formation in a concentration-dependent manner. HT at 50 and 100 μmol/L concentrations increased the GSH level. The specific DNA-binding activities of NF-κB in nuclear extracts from 50 and 100 μmol/L HT treatments were significantly suppressed. The antioxidant N-acetylcysteine （NAC） also showed the same effects as HT on LPS-induced ROS and NO production, change of GSH level, and NF-κB activation. Conclusion： These findings suggest that HT has antioxidant activity by suppressing intracellular oxidative stress and NF- κB activation in THP-1 cells.