三七总皂苷调节iNOS-NO-NF-κB信号通路抑制LPS诱导的RAW246.7细胞炎症研究  

Inhibitory functions of Panax notoginseng saponins on LPS-induced RAW264.7 cells inflammation via iNOS-NO-NF-kB signaling pathways

作　　者：冯晓异 赵微 何朋伦 陈文慧 姚政 FENG Xiaoyi;ZHAO Wei;HE Peng lun;CHEN Wenhui;YAO Zheng(Yunnan University of Chinese Medicine,Kunming 650500,China)

机构地区：[1]云南中医药大学,云南昆明650500

出　　处：《药物评价研究》2020年第4期670-675,共6页Drug Evaluation Research

基　　金：国家自然科学基金资助项目(81760818、81160492);云南省应用基础研究计划项目-中医联合专项[2017FF11(-013)]。

摘　　要：目的探讨三七总皂苷(PNS)对脂多糖(LPS)诱导的RAW264.7细胞iNOS-NO-NF-κB信号通路相关分子表达及活性的影响。方法MTT比色法检测PNS对RAW264.7细胞增殖的抑制作用;不同浓度PNS(25、50、100μg/mL)干预体外培养LPS诱导的RAW264.7细胞24、48 h后,Griess法检测NO变化;PNS干预24 h后,Western blotting检测iNOS、NF-κB、p-NF-κB、IKKα、p-IKKα、p-ΙκΒα蛋白表达量的变化;PNS干预4 h后,激光共聚焦显微镜检测NF-κB转位入核情况。结果PNS浓度大于150μg/mL时,才表现出显著抑制细胞增殖的作用。25、50、100μg/mL PNS作用于RAW264.7细胞24和48 h后,与模型组比较,NO生成量均显著降低(P<0.001)。50、100μg/mL PNS作用于RAW264.724 h后,iNOS、NF-κB蛋白表达量显著降低,磷酸化蛋白p-NF-κB、p-IKKα、p-ΙκΒα表达水平也显著降低(P<0.01、0.001)。与模型组比较,PNS 25、50、100μg/mL组核因子p65入核荧光强度显著降低(P<0.01、0.001)。结论PNS能够显著抑制LPS诱导的RAW264.7细胞iNOS-NO-NF-κB信号通路的活性,降低细胞炎症反应。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(25-100μg/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-IκBαby Western blot,the NF-κB p65 translocation by confocal laser scanning fluorescence microscopy.Results When the concentration of PNS was more than 150μg/mL,the inhibition of cell proliferation was significant.25,50,100μg/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 100μg/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-IκBαproteins were all decreased in a dose-dependment manner(P<0.01,0.001).Conclusion Taken together,PNS exerts anti0 inflammatory effects,by inhibiting the activity of iNOS-NO-NF-κB signaling pathway in LPS-treated RAW264.7 macrophages.

关 键 词：三七总皂苷 RAW264.7 NO通路 NF-ΚB通路 炎症 

分 类 号：R285.5[医药卫生—中药学]