4种迷迭香化合物对脂多糖诱导的RAW264.7细胞氧化应激和炎症的抑制作用

Inhibitory Effect of Four Bioactive Compounds from Rosemary on Lipopolysaccharide(LPS)-Induced Oxidative Stress and Inflammation in RAW264.7 Cells

目的:研究4种迷迭香化合物(鼠尾草酸、鼠尾草酚、迷迭香酚和迷迭香酸)对脂多糖(lipopolysaccharide,LPS)诱导的RAW264.7细胞氧化应激和炎症反应的抑制作用。方法:以LPS诱导RAW264.7细胞构建炎症模型,通过比色法、实时荧光定量聚合酶链式反应、Western blot等方法表征迷迭香化合物对活性氧(reactive oxygen species,ROS)、炎症因子以及相关酶类的影响。结果:上述4种迷迭香化合物对LPS诱导的细胞内ROS相对含量、丙二醛(malonic dialdehyde,MDA)含量、NO释放量和肿瘤坏死因子α(tumor necrosis factor α,TNF-α)、白细胞介素1β(interleukin 1β,IL-1β)、IL-6 mRNA相对表达量,以及诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和环氧合酶2(cyclooxygenase 2,COX-2)蛋白相对表达量的升高有显著抑制作用,对超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活力有显著保护作用,且具有浓度依赖效应。质量浓度10μg/mL的鼠尾草酸对LPS诱导的ROS相对含量、NO释放量、TNF-α mRNA相对表达量、MDA含量升高的抑制作用和对SOD和CAT活力的保护作用更好,质量浓度10μg/mL的迷迭香酚对LPS诱导的IL-1β、IL-6 mRNA相对表达量、iNOS和COX-2蛋白相对表达量升高的抑制作用更好。结论:4种迷迭香化合物均具有一定的抗氧化和抗炎作用,能缓解炎症反应,其中水溶性迷迭香酸有效质量浓度高于其他3种。

Objective: Our aim is to investigate the inhibitory effects of four bioactive compounds from rosemary(carnosic acid, carnosol, rosmanol and rosmarinic acid) against oxidative stress and inflammatory response in lipopolysaccharide(LPS)-induced RAW264.7 cells. Methods: The effects of these compounds on reactive oxygen species(ROS), inflammatory factors and related enzymes were characterized by various methods including colorimetry, fluorescence quantitative polymerase chain reaction(PCR) and Western blot. Results: All four compounds had significant inhibitory effects on the expression or production of ROS, malonic dialdehyde(MDA), nitric oxide(NO), tumor necrosis factor-α(TNF-α),interleukin(IL)-1β, IL-6, inducible nitric oxide synthase(iNOS) and cyclooxygenase 2(COX-2) in LPS-induced cells. They had a significant protective effect on the activity of superoxide dismutase(SOD) and catalase(CAT) in a concentrationdependent manner. Carnosic acid at 10 μg/mL had a better inhibitory effect on the LPS-induced increases in relative ROS levels, NO release, TNF-α mRNA expression and MDA levels, and a better protective effect on SOD and CAT activity.Rosmanol at 10 μg/mL had a better inhibitory effect on the LPS-induced increases the relative mRNA expression levels of IL-1β and IL-6 and the relative protein expression levels of iNOS and COX-2. Conclusion: All four compounds from rosemary have antioxidant and anti-inflammatory effects and alleviates inflammatory responses. The effective concentration of water-soluble rosmarinic acid is significantly higher than that of the other three compounds.