黄芪总黄酮体外抗炎作用及对MAPKs信号通路的调控  

Anti-inflammatory effect of total flavonoids of Astragalus and its regulation on MAPKs signaling pathway in vitro

作　　者：周鸿缘 张贤 王萌[1] 葛冰洁 王政[1] 李海涛 张雪梅[1] ZHOU Hong-yuan;ZHANG Xian;WANG Meng;GE Bing-jie;WANG Zheng;LI Hai-tao;ZHANG Xue-mei(Agricultural College of Yanbian University,Yanji,Jilin 133002,China;Institute of Special Animal and Plant Sciences of CAAS,Changchun 130112,China)

机构地区：[1]延边大学农学院,吉林延吉133002 [2]中国农业科学院特产研究所,吉林长春130112

出　　处：《中国兽医学报》2020年第12期2392-2397,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31660727,31260622);吉林省科技厅重点科技攻关资助项目(20160204016NY)。

摘　　要：以LPS诱导的小鼠腹腔巨噬细胞RAW264.7为模型,探讨黄芪总黄酮(TFA)的体外抗炎作用及其机制。MTT法测定TFA对RAW264.7细胞的毒性;在安全浓度下设立TFA低、中、高剂量组以及空白组和模型组,Griess法检测细胞上清液炎性介质NO的分泌量,ELISA法测定细胞上清液炎性介质PGE2和炎性因子IL-1β、IL-6、TNF-α和IFN-γ的含量,RT-PCR法测定细胞iNOS和COX-2 mRNA的含量,Western blot法测定iNOS和COX-2蛋白表达情况,同时分析MAPKs信号通路的关键蛋白p38、ERK1/2和JNK磷酸化水平的变化规律。结果显示,TFA可呈剂量依赖性的抑制LPS诱导所致RAW264.7细胞NO、PGE2、IL-1β、IL-6、TNF-α和IFN-γ的过度分泌,下调iNOS和COX-2 mRNA及其蛋白的表达,抑制MAPKs信号通路关键蛋白p38和JNK的过度磷酸化。结果表明,TFA可通过调控RAW264.7细胞MAPKs相关通路以及iNOS和COX-2表达,进而抑制炎性介质NO、PGE2和炎性因子IL-1β、IL-6、TNF-α、IFN-γ生成,发挥其体外抗炎的作用。The in vitro anti-inflammatory effect and its mechanism of total flavonoids of Astragalus(TFA)were studied using LPS induced mouse peritoneal macrophage RAW264.7 cells as a model.Toxicity of TFA to RAW264.7 cells was determined by MTT method.The low-,medium-and high doses of TFA groups,blank group and model group were set up.Griess method was used to detect the secretion of inflam matory mediator NO,and ELISA method was used to detect the contents of inflammatory mediator PGE2 and inflammatory cytokines IL-1β,IL-6,TNF-α and IFN-γ.RT-PCR method was used to detect the content of iNOS and COX 2 mRNA.The changes in the phosphoryl-ation levels of key proteins in MAPKs signaling pathway,such as iNOS,COX 2,p38,ERK1/2 and JNK were also analyzed by Western blot method.The results showed that TFA dose-dependently inhibited LPS induced oversecretion of NO,PGE2,IL-1β,IL-6,TNF-α and IFN-γ,and TFA down-regulated the expression of iNOS and COX-2 mRNA and protein,and suppressed the overphospho-rylation of p38 and JNK in MAPKs signaling pathway.The results indicate that TFA can inhibit the production of inflammatory mediators NO,PGE2 and inflammatory cytokines IL-1β,IL-6,TNF-α,IFN-γ by regulating the MAPKs-related pathway and the expression of iNOS and COX-2 in RA W264.7 cells,thus exerting its anti-inflammatory effects in vitro.

关 键 词：TFA RAW264.7细胞 体外抗炎 MAPKS信号通路 

分 类 号：S859.79[农业科学—临床兽医学]