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作 者:郭毅斌[1] 曹红卫[1] 陈莉萍[2] 鲁永玲[1] 郑江[1] 肖光夏[3]
机构地区:[1]第三军医大学西南医院综合实验研究中心,重庆400038 [2]第三军医大学西南医院全军肝胆外科研究所,重庆 400038 [3]第三军医大学西南医院全军烧伤研究所,创伤.烧伤与复合伤国家重点实验室,重庆 400038
出 处:《中华烧伤杂志》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)
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.
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