摘要
目的 了解白鲜皮提取物2(DPR-2)对内毒素/脂多糖(LPS)的体外拮抗活性。方法 采用动态比浊法鲎试验定量检测DPR-2对LPS(0.1μg/L)的中和作用,激光共聚焦扫描显微镜观察不同浓度DPR-2(0、8、0、16.0、32.0、64.0mg/L)对异硫氰酸荧光素-LPS(FITC-LPS,100.0μg/L)与小鼠单核细胞株RAW264、7结合力的影响,用荧光定量反转录.聚合酶链反应(RT-PCR)法检测以上浓度DPR-2对LPS(100.0μg/L)刺激的RAW264.7细胞肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)mRNA表达的影响。结果 DPR-2对LPS具有一定的中和作用(P〈0.05或P〈0.01);浓度为32.0、64.0mg/L时可显著抑制FITC-LPS与RAW264.7细胞结合(P〈0.01);对LPS刺激的小鼠RAW264.7细胞TNF-α和IL-6mRNA的表达有抑制作用,该作用具有一定的剂量-效应关系。结论 DPR-2可通过中和LPS的作用阻断与RAW264.7细胞膜受体结合,从而抑制LPS介导的细胞活化。
Objective To investigate the lipopolysaccharide (LPS) antagonizing biological activity of densefruit pittany root-bark extract (DPR-2) in vitro. Methods The effect of DPR-2 in neutralizing LPS (0. 1μg/L) was detected by kinetic turbidimetric limulus test. The effect of different concentrations of DPR-2 (0,8.0,16, 0,32, 0,64. 0 mg/L) on binding of FITC-conjugated LPS ( FITC-LPS, 100.0 μg/L) to murine RAW264.7 cells was analyzed with laser scanning confocal microscopy. The expression of TNF-α and IL-6 mRNA in RAW264.7 cells after exposure to LPS ( 100.0 μg/L) were determined by real-time RTPCR. Results DPR-2 could neutralize LPS( P 〈 0.05 or P 〈 0.01 ), and inhibit the binding of FITC-LPS to RAW264.7 ceils in a dose-dependent manner when the concentration of DPR-2 was above 16.0 mg/L. Furthermore, DPR-2 could markedly inhibit the expression of TNF-α and IL-6 mRNA in LPS-stimulated murine RAW264.7 cells. Conclusion DPR-2 exhibit an anti-LPS effect in vitro, which may be related to its capacity to neutralize LPS and inhibit binding of LPS for its receptors.
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2007年第2期104-107,共4页
Chinese Journal of Burns
基金
国家自然科学基金(30371767)
关键词
内毒素类
脂多糖类
脓毒症
白鲜皮
Endotoxins
Lipopolysaccharide
Sepsis
Cortex dictamni