竹节参醇提物对LPS诱导RAW264.7细胞炎症的保护作用

Protective Effect of Panax japonicus Ethanol Extract on the Inflammation of RAW264. 7 Cells Induced by LPS

目的:观察竹节参醇提物对脂多糖(LPS)刺激小鼠单核巨噬白血病细胞RAW264.7细胞炎症保护作用的初步探讨。方法:用不同质量浓度的竹节参醇提物处理RAW264.7细胞,噻唑蓝(MTT)法检测细胞活性;检测LPS刺激的RAW264.7细胞一氧化氮(NO)释放量以及竹节参醇提物干预后RAW264.7细胞NO释放量;酶联免疫吸附(ELISA)法检测肿瘤坏死因子-α(TNF-α)、白介素1β(IL-1β)分泌;聚合酶链反应(PCR)法检测诱导型一氧化氮合酶(iNOS)mRNA的表达;免疫印迹(Western blot)法检测胞浆胞核核转录因子-κB(NF-κB)蛋白表达。结果:竹节参醇提物作用RAW264.7细胞的安全范围<100 mg·L-1;LPS的用药质量浓度为1 mg·L-1;与LPS模型组相比,竹节参醇提物0.1,1,10,40 mg·L-1能有效抑制NO释放(抑制率分别为31.2%,41%,46.1%,55.2%)和抑制TNF-α,IL-1β的分泌(P<0.05或P<0.01);竹节参醇提物10,40 mg·L-1能下调LPS模型组iNOS mRNA表达且抑制NF-κB的核移位。结论:竹节参醇提物对LPS刺激的RAW264.7细胞炎症具有保护作用,其保护机制可能与调控NF-κB通路,进而抑制NO的释放,降低TNF-α,IL-1β,iNOS表达有关。

Objective： To observe the protective effect of Panax japonicus ethanol extract on the inflammation of mice mononuclear macrophage RAW264.7 cells induced by lipopolysaccharide（LPS）. Method： The RAW264.7 cells were treated with different concentration of P. japonicus ethanol extract, cell viability was determined with MTT method; the nitric oxide（NO） release of RAW264.7 cells stimulated by LPS and treated with LPS and P. japonicus ethanol extract was detected with Griess. ELISA were used to detect the expression of Tumor necrosis factor-α（TNF-α）, interleukin 1β（IL-1β）; nitric oxide synthase mRNA （iNOS mRNA） and neclear factor-κB（NF-κB） was tested by PCR and Western bolt respectively. Result： The safe range of ethanol extract of P. japonicus is less than 100 mg·L-1; Concentration of LPS was 1 mg·L-1;Compared with the LPS model group, P. japonicus ethanol extract 0.1, 1, 10, 40 mg·L-1 could inhibit the of NO effectively, the inhibition rates were 31.2%, 41%, 46.1%, 55.2%, and inhibit the release of TNF-α, IL-1β, with significant difference（P〈0.05 or P〈0.01）;P. japonicus ethanol extract 10, 40 mg·L-1 could down-regulate the expression of iNOS mRNA and inhibit NF-κB nuclear translocation. Conclusion： P. japonicus ethanol extract has protective effects on RAW264.7 cells stimulated by LPS, the mechanism of P. japonicus ethanol extract may be related to it＇s regulation on NF-κB signal pathway, thereby inhibiting the release of NO and reducing the expression of TNF-α, IL-1β and iNOS.