水飞蓟宾-磷脂酰胆碱复合物对脂多糖诱导小鼠巨噬细胞核因子-κB活化及核因子-κB抑制蛋白α磷酸化的影响

Inhibitory Effect of Silybin-Phosphatidylcholine Compound on the Activation of Nuclear Factor-κB and Phosphorylation of Inhibitors of Nuclear Factor-κB α in Lipopolysaccharide Induced by Mouse Phagocyte

目的研究水飞蓟宾-磷脂酰胆碱复合物（SPC）对脂多糖（LPS）诱导小鼠巨噬细胞核因子-κB（NF-κB）活化及NF-κB抑制蛋白α（IκBα）磷酸化的影响。方法提取健康6～8周龄昆明种小鼠腹腔巨噬细胞，培养成活后调细胞水平为2×10^5mL^-1，对照组加：入等量9g／L盐水，LPS组加入LPS，使其终水平10μg／mL，LPS刺激细胞24h，SPC干预组加入不同水平SPC，干预细胞2h后加入终水平为10μg／mL LPS刺激24h。免疫细胞化学法观测NF—κB激活、IκBα磷酸化的表达。结果对照组小鼠巨噬细胞细胞核内NF-κB p65水平很低，LPS组细胞核NF-κB p65水平显著高于对照组〈P〈0．01），不同水平SPC干预组小鼠巨噬细胞NF—κB p65水平降低，即NF—κB p65表达减少。且呈水平依赖性降低（P〈0．01）。细胞质内磷酸化的IκBα表达同胞核NF-κB p65水平变化基本一致。结论，SPC抑制LPS诱导的小鼠巨噬细胞内NF—κB活化，可能是通过抑制IκBα磷酸化及降解途径完成。

Objective To explore the effects of silybin -phosphatldyleholine compound （ SPC ）on lipopolysaccharide （LPS） - induced activation of nuclear factor - κB （NF - κB） and phosphorylation and degradation of inhibitors of NF-κBα（IκBα）. Methods ： Phagecyte were collected in abdominal cavity of Kunming mousse aged ：6 to 8 weeks, Cultured phagocyte （2× 10^5 mL^-1 ） Were divrded into, control;LPS and SPC groups randomly, phagocyte in control group were added into the ：same volume 0.9g/L sodium chloride. Phagocyte in LPS group were added into a single bolas of LPS（ 10μg/mL LPS） for 24 hours, phagocyte in SPC groups were preincubated with different concentration of SPC for 2 hours followed by a 24 hours incubation with 10μg/mL LPS. immanocytochemistry were used to measure the contents of NF-κB, phosphorylated IκBα in phagocyte. Results The content of NF- κB p65 located in the nuclear in control group was little. The contant of NF - κB. p65 located in the nuclear in LPS group markly higher： than that in control group（ P 〈 0.01 ）. When pretreated with different, coneentrations of SPC for 2 hours., the content of NF - κB p65 located in the nuclear decreased gradually in a dose - dependent（P 〈0.01 ）. The statistical analysis change trend ofphosphorylated IκBα located in the cytoplasm was almost accord with NF - κB p65. Conclusion The expression of NF-κB Was significantly inhibited by SPC, and this effect was mediated through the inhibition of the phosphorylation and degradation of IκBα.