摘要
目的研究冬凌草甲素对脂多糖(LPS)诱导的Raw264.7巨噬细胞炎症反应的影响。方法选用小鼠巨噬细胞Raw264.7,CCK-8法确定冬凌草甲素作用的最适浓度;设立正常对照组、模型对照组(LPS组)、实验组(冬凌草甲素预处理+LPS组)和阳性药组(地塞米松预处理+LPS组),实时定量PCR法检测TNF-α、IL-1β、IL-6、IL-10和TLR4 mRNA表达水平的改变;Western blot法检测NF-κB p65、磷酸化p65(p-p65)蛋白表达水平的改变。结果冬凌草甲素作用于Raw264.7细胞的最佳浓度为10μmol/L;与LPS组比较,冬凌草甲素预处理实验组Raw264.7细胞内TNF-α、IL-1β和IL-6 mRNA表达水平明显下降,IL-10 mRNA表达水平明显升高,TLR4基因表达降低,NF-κB活化入核减少。结论冬凌草甲素能下调LPS诱导的Raw264.7细胞促炎因子表达,其抗炎免疫作用机制与抑制TLR4-NF-κB信号通路的活化有关。
Objective To investigate the effect of oridonin on lipopolysaccharide （LPS）-induced inflammation response in Raw264.7 macrophage cell line. Methods The optimal concentration of oridonin on Raw264.7 maerophages was determined by CCK8 method. The inflammation models were established by LPS stimulation and divided into four groups including the normal control, the LPS group, the experimental group （oridonin pretreatment ＋ LPS） and the positive control （dexamethasone pretreatment＋LPS）. The changes of TNF-α, IL-1β, IL-6, IL-10 and TLR4 mRNA levels were measured by real-time quantitative PCR. The expression of NF-κB p65 and phosphorylated p65 （p-p65） were detected by western blot. Results At the optimal concentration （ 10 μmol/L）, oridonin pretreatment significantly inhibited the expression of TNF-α, IL-1β and IL-6 mRNA induced by LPS and increased the expression of IL-10 mRNA in Raw264.7 cells. Oridonin also inhibited the expression of TLR4 and activation of NF-KB translocation into nucleus. Conclusion Oridonin can inhibit the expression of proinflammatory cytokines induced by LPS and exert anti-inflammation immune mechanism through TLR4-NF-κB signaling pathway.
出处
《广东药科大学学报》
CAS
2017年第2期250-254,共5页
Journal of Guangdong Pharmaceutical University
基金
国家自然科学基金项目(31471290)
广东省科技计划项目(2013B021800082)
