畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (3): 609-614.doi: 10.11843/j.issn.0366-6964.2016.03.026

• 临床兽医 • 上一篇    下一篇

金英黄归汤对LPS介导乳腺上皮细胞TLR4信号通路中相关因子的影响

李欣1,2,李中改1,2,张小艺1,2,周送桂1,3,王玉坤1,2,冯将1,2,易琼1,王鲁1*   

  1. (1.贵州大学 贵州省生化工程中心,贵阳 550025;2.贵州大学 动物科学学院,贵阳 550025;3.贵州大学 药学院,贵阳 550025)
  • 收稿日期:2015-08-12 出版日期:2016-03-23 发布日期:2016-03-23
  • 通讯作者: 王鲁(1970-),教授,博士,主要从事中兽医学及中药药理学研究,E-mail:wanglu7007@163.com
  • 作者简介:李欣(1991-),男,吉林通化人,硕士生,主要从事中药药理学研究,E-mail:690079602@qq.com
  • 基金资助:

    国家自然科学基金项目(31260618;31470128);北京农学院兽医学(中医药)北京市重点实验室开放课题

Effects of Jin-Ying-Tang on Correlated Molecules of Toll-like Receptor 4 Signaling of LPS-Induced Mouse Mammary Epithelial Cells in vitro

LI Xin1,2,LI Zhong-gai1,2,ZHANG Xiao-yi1,2,ZHOU Song-gui1,3,WANG Yu-kun1,2,FENG Jiang1,2,YI Qiong1,WANG Lu1*   

  1. (1.Biochemistry Engineering Center of Guizhou Province,Guizhou University,Guiyang 550025,China;2.College of Animal Science,Guizhou University,Guiyang 550025,China;3.College of Pharmacy,Guizhou University,Guiyang 550025,China)
  • Received:2015-08-12 Online:2016-03-23 Published:2016-03-23

摘要:

为了评价金英黄归汤抗LPS致炎的作用机制,作者采用ELISA法检测金英黄归汤对细胞分泌细胞因子TNF-α、IL-8的影响,采用qPCR法研究药物对TLR4通路中IL-1受体相关激酶-1(IRAK-1)、肿瘤坏死因子受体6(TRAF-6)、转化生长因子激活激酶(TAK-1)、NF-κB诱导激酶(NIK)、抑制性NF-κB(IκB) mRNA的转录影响。结果显示,金英黄归汤低剂量组(39.1 μg•mL-1)、中剂量组(391 μg•mL-1)、高剂量组(3 910 μg•mL-1)能显著(P<0.05)或极显著(P<0.01)减少LPS介导的TNF-α和IL-8分泌;能显著(P<0.05)或极显著(P<0.01)减少LPS介导的IRAK-1、TRAF-6、TAK-1、IκBNIK mRNA的转录。结果提示,金英黄归汤通过抑制小鼠MECs 的TLR4/NF-κB信号通路中IRAK-1/TRAF-6/TAK-1/NIK途径的活化来控制炎性因子的释放而发挥抗炎作用,而不是通过IRAK-1/TRAF-6/TAK-1/IκB途径实现的,这可能是金英黄归汤的抗炎机制。

Abstract:

The study was conducted to investigate the effects of Jin-Ying-Tang (JYT) on correlated molecules of TLR4NF-κB signaling of LPS-stimulated mouse mammary epithelial cells(MECs) in vitro and explore its underlying molecular mechanisms.ELISA was performed to detect changes of the content of IL-8 and TNF-α in the culture supernatants.The mRNA transcription of TLR4-NF-κB signaling molecules such as IL-1 receptor-associated kinase-1(IRAK-1),tumour necrosis factor receptor associated factor 6(TRAF-6),Transforming Growth Factor activated kinase 1(TAK-1),NF-κB inducing kinase(NIK),Inhibitor of κB(IκB) were determined by qRT-PCR.The results showed that JYT could significantly decrease the expression of IL-8 and TNF-α in LPS-stimulated MECs(P<0.05,P<0.01);what’s more,JYT could down-regulate the expression of IRAK-1,TRAF-6,TAK-1,IκBNIK mRNA activated by LPS(P<0.05,P<0.01).It is concluded that JYT attenuates LPS-induced inflammatory response in MECs by inhibiting the activitation of IRAK-1/TRAF-6/TAK-1/NIK pathway,but not of IRAK-1/TRAF-6/TAK-1/IκB pathway.

中图分类号: