化合物W3D通过调控TLR4-MyD88-NF-κB通路抑制LPS诱导的RAW264.7细胞炎症因子的释放  

Inhibitory effect of compound W3D on LPS-induced release of inflammatory mediators of RAW264. 7 cells through TLR4-MyD88-NF-κB signaling pathway

作　　者：罗捷然 赵蓓 唐莉[1] 葛睿[1] 李青山[1,2] LUO Jie-ran;ZHAO Bei;TANG Li;GE Rui;LI Qing-shan(School of Pharmaceutical Science,Shanxi Medical University,Taiyuan030001,China;School of Traditional Chinese medicine,Shanxi University of Chinese Medicine,Taiyuan 030024,China)

机构地区：[1]山西医科大学药学院,山西太原030001 [2]山西中医药大学中药学院,山西太原030024

出　　处：《中国药理学通报》2018年第7期977-982,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81602976);山西省青年科技研究基金(No 201601D021159);山西省回国留学人员重点科研资助项目(No 2014-重点2);山西省高等学校科技创新项目(No 2014132;2015150);山西医科大学博士启动金(No 03201319)

摘　　要：目的建立脂多糖(LPS)诱导的小鼠单核巨噬细胞(RAW264.7)炎症模型,探究新型苯并噁唑酮衍生物4-(5'-二甲氨基)-萘磺酰氧基苯并噁唑酮(W3D)的抗炎活性及其对TLR4-MyD88-NF-κB通路的调控作用。方法 MTT法测定化合物W3D对细胞活力的影响;LPS与不同浓度的化合物W3D共同作用RAW264.7细胞后,ELISA法测定细胞上清液中TNF-α、IL-6、IL-1β、COX-2的含量,Western blot法检测IL-6、TLR4、MyD88、IRAK4、NF-κB的蛋白表达;实时荧光定量PCR法检测细胞中TLR4、MyD88、IL-6 mRNA的表达。结果化合物W3D对LPS诱导的RAW264.7细胞培养液中炎症因子TNF-α、IL-6、IL-1β的分泌有明显的抑制作用,但对COX-2无抑制活性;可明显下调TLR4、MyD88、IL-6的蛋白与mRNA的表达,抑制IRAK4磷酸化和NF-κB的入核活化。结论化合物W3D可通过调控TLR4-MyD88-IRAK4-NF-κB信号通路,抑制TNF-α、IL-6、IL-1β等炎症因子的释放而发挥抗炎活性。Aim To investigate the effect of the novel benzoxazolone derivative 4-（ 5＇-dimethylamino）-naphthalenesulfonyl-2（ 3H）-benzoxazolone（ W3D） on TLR4-MyD88-NF-κB signaling pathway in LPS-induced RAW264.7 cells.Methods The cell viability was detected by MTT assay,and the contents of TNF-α,IL-6,IL-1β and COX-2 in the cell supernatant were analyzed using ELISA methods.The protein expression of IL-6,TLR4,MyD88,p-IRAK4 and NF-κB were investigated by western blot analysis, and the mRNA expressions of TLR4,MyD88 and IL-6 were analyzed by RT-PCR.Results W3D could obviously inhibit the production of TNF-α,IL-6 and IL-1β in LPSinduced RAW264.7 cell supernatant,but it had no effect on the release of COX-2.Compared with the model group,the expressions of TLR4,MyD88 and IL-6 were decreased significantly in a dose dependent manner.Meanwhile,the expressions of p-IRAK4 and nucleus of NF-κB were decrease in W3D treated group compared with the model group.Conclusion The novel compound W3D could inhibit the release of the inflammatory mediators through the regulation of TLR4-MyD88-NF-κB signaling pathway.

关 键 词：苯并噁唑酮衍生物W3D RAW264.7细胞 炎症 TLR4 MYD88 NF-ΚB 炎症因子 

分 类 号：R-33[医药卫生] R329.2