水飞蓟宾-磷脂酰胆碱复合物对小鼠巨噬细胞一氧化氮及活性氧生成的抑制作用  

Inhibitory Effect of Silybin-Phosphatidylcholine Compound on Production of Nitric Oxide and Reactive Oxygen Species in Mouse Macrophage

作　　者：郭蕴琦[1] 郭蕴岚[2] 刘昕慧[3] 裴利宏[4] 

机构地区：[1]新乡医学院第一附属医院药剂科,河南卫辉453100 [2]新乡医学院第三附属医院药剂科,河南新乡453003 [3]新乡医学院第二附属医院精神科,河南新乡453002 [4]新乡医学院第一附属医院检验科,河南卫辉453100

出　　处：《实用儿科临床杂志》2008年第21期1691-1692,共2页Journal of Applied Clinical Pediatrics

摘　　要：目的研究水飞蓟宾-磷脂酰胆碱复合物(SPC)对过氧化氢(H2O2)诱导小鼠巨噬细胞内一氧化氮(NO)及活性氧(ROS)生成的影响。方法健康6～8周龄昆明种小鼠,腹腔提取巨噬细胞,培养成活后调细胞水平至2×108L-1,空白对照组加入等量的9g/L盐水,H2O2组加入H2O2,使其终质量浓度为1mmol/L刺激细胞30min,SPC干预组加入不同水平SPC干预细胞2h后加入终质量浓度为1mmol/LH2O2刺激30min。免疫细胞化学法观察诱导型一氧化氮合酶(iNOS)的表达,Griess法测定上清液NO的生成量,荧光探针DCFH-DA法测定巨噬细胞内ROS生成。结果H2O2组上清液NO2-水平(相当于NO累积生成量)显著高于对照组(P<0.01),不同水平SPC干预组NO2-水平降低,即NO表达减少,且呈质量浓度依赖性降低(P<0.05)。各组细胞质内iNOS表达同上清液NO的生成量呈现相应的变化趋势。H2O2组小鼠巨噬细胞内ROS显著升高,与对照组比较有显著性差异(P<0.01),不同水平SPC预先干预2h后,ROS生成量呈质量浓度依赖性降低(P<0.01)。结论SPC可能是通过抑制iNOS表达来抑制H2O2诱导的小鼠巨噬细胞内NO的生成,另外SPC也可抑制小鼠巨噬细胞内ROS生成,从而减轻组织细胞的氧化损伤。Objective To investigate the effects of silybin - phosphatidylcholine compound （SPC） on H2O2 - induced the production of nitric oxide （NO） and reactive oxygen species （ROS） in mouse macrophage. Methods Macrophage were collected in abdominal cavity of 6 -8 week Kunming mouse, and cultured macrophage （2 ×10^8 L^-1 ） were divided into control and SPC group randomly. Macrophage in control group were added into the same volume（0.1 mL） of 9 g/L sodium chloride. Macrophage in H2O2 group were added into a single bolus of H2O2 （1 mmol/L H2O2 ） for 30 min ,while macrophage in SPC group were preincubated with different concentration of SPC for 2 h followed by a 30 min incubation with 1 mmol/L H2O2. Immunocytochemistry were used to measure the contents of inducible nitric - oxide synthase （ iNOS）, NO production was determined by Griess reactive, ROS was determined by DCFH - DA Fluorescence probe. Results The production of NO in H2O2 group markedly higher than that in control group （P 〈 0.01 ）, when pretreated with different concentrations of SPC for 2 h. The production of NO in isolation supernatants decreased gradually in a dose- dependent（ P 〈 0.05 ）. The statistical analysis of change trend of iNOS located in the cytoplasm was almost in accord with the production of NO. Conclusions The production of NO in mouse macrophage is significantly inhibited by SPC, and this effect is mediated through the inhibition of the expression of iNOS. Otherwise, the production of ROS in mouse macrophage is significantly inhibited by SPC, which make oxidative damage of histiocyte decreased.

关 键 词：水飞蓟宾 水飞蓟宾-磷脂酰胆碱复合物 诱导型一氧化氮合酶 一氧化氮 活性氧 

分 类 号：R285.5[医药卫生—中药学]