目的 探討三七總皂苷（PNS）對脂多糖（LPS）誘導的RAW264.7細胞iNOS-NO-NF-κB信號通路相關分子表達及活性的影響。方法 MTT比色法檢測PNS對RAW264.7細胞增殖的抑制作用；不同濃度PNS（25、50、100 μg/mL）干預體外培養(yǎng)LPS誘導的RAW264.7細胞24、48 h后，Griess法檢測NO變化；PNS干預24 h后，Western blotting檢測iNOS、NF-κB、p-NF-κB、IKKα、p-IKKα、p-ΙκΒα蛋白表達量的變化；PNS干預4 h后，激光共聚焦顯微鏡檢測NF-κB轉位入核情況。結果 PNS濃度大于150 μg/mL時，才表現出顯著抑制細胞增殖的作用。25、50、100 μg/mL PNS作用于RAW264.7細胞24和48 h后，與模型組比較，NO生成量均顯著降低（P<0.001）。50、100 μg/mL PNS作用于RAW264.7 24 h后，iNOS、NF-κB蛋白表達量顯著降低，磷酸化蛋白p-NF-κB、p-IKKα、p-ΙκΒα表達水平也顯著降低（P<0.01、0.001）。與模型組比較，PNS 25、50、100 μg/mL組核因子p65入核熒光強度顯著降低（P<0.01、0.001）。結論 PNS能夠顯著抑制LPS誘導的RAW264.7細胞iNOS-NO-NF-κB信號通路的活性，降低細胞炎癥反應。

Objective To investigate the effect of Panax notoginseng saponins (PNS) on the expression and activity of iNOS-NONF-κB signaling pathway related molecules in lipopolysaccharide (LPS)-stimulated RAW264.7 cells in vitro. Methods RAW264.7 cells were treated with different concentrations (25-100 μg/mL) of PNS for different periods of time (24-48 h). Antiinflammation effects of PNS in LPS-stimulated RAW264.7 macrophages were measured based on the detection of nitric oxide (NO) overproduction (Griess method, DAF-FM DA fluorescence assay and NO₂^- scavenging assay), and the cell proliferation was measured by MTT assay, the expression of iNOS, NF-κB, p-NF-κB, IKKα, p-IKKα, p-IκBα by Western blot, the NF-κB p65 translocation by confocal laser scanning fluorescence microscopy. Results When the concentration of PNS was more than 150 μg/mL, the inhibition of cell proliferation was significant. 25, 50, 100 μg/mL PNS treated RAW264.7 cells for 24 and 48 h, compared with the model group, NO production was significantly reduced (P<0.001). After PNS (25, 50 and 100 μg/mL) was applied to RAW264.7 cells for 24 h, the expression of iNOS, NF-κB proteins and the expression of p-NF-κB, p-IKKα, p-IκBα proteins were all decreased in a dose-dependment manner (P<0.01, 0.001). Conclusion Taken together, PNS exerts anti0inflammatory effects, by inhibiting the activity of iNOS-NO-NF-κB signaling pathway in LPS-treated RAW264.7 macrophages.

R285.5

國家自然科學基金資助項目（81760818、81160492）；云南省應用基礎研究計劃項目-中醫(yī)聯合專項[2017FF11（-013）]