Wang Guang-Qing, Yang Xiao-Yan, Jia Yi-Tao, Xia Zhao-Fan
Institute of Burn Surgery, Changhai Hospital, Second Military Medical University, 174 Changhai Road, Shanghai 200433, People's Republic of China.
Inflammation. 2009 Aug;32(4):265-9. doi: 10.1007/s10753-009-9129-z.
The purpose of present study was to explore the possibility of Tec kinase as a mediator for IL-8 transcription in monocytes stimulated with LPS. Plasmids of mouse Tec kinase IV or Tec kinase IV with inactivating point mutations generated with QuikChange site-directed mutagenesis were co-transfected with IL-8 promoter driven luciferase construct into RAW264.7 cells, then luciferase activity was measured with a luminometer. The results shown Tec kinase could significantly enhance IL-8 transcription. Furthermore, point inactivating mutation in SH2, PH or PTK domain almost completely abolish the effects of Tec kinase on the transcription of IL-8. In the transfection experiment, PD98059, a MEK1 inhibitor, decreased the transcription of IL-8 in a dose dependent pattern. When siRNA for Tec kinase was transfected into THP-1 cells, it could efficiently block the production of IL-8 from THP-1 cells (p < 0.01) stimulated with LPS. In conclusion, Tec kinase may mediate the transcription of IL-8 in monocyte stimulated with LPS.
本研究的目的是探讨Tec激酶作为脂多糖刺激单核细胞中白细胞介素-8（IL-8）转录介质的可能性。将通过QuikChange定点诱变产生的小鼠Tec激酶IV或具有失活点突变的Tec激酶IV质粒与白细胞介素-8启动子驱动的荧光素酶构建体共转染到RAW264.7细胞中，然后用光度计测量荧光素酶活性。结果表明，Tec激酶可显著增强白细胞介素-8的转录。此外，SH2、PH或PTK结构域中的点失活突变几乎完全消除了Tec激酶对白细胞介素-8转录的影响。在转染实验中，MEK1抑制剂PD98059以剂量依赖性方式降低白细胞介素-8的转录。当将Tec激酶的小干扰RNA（siRNA）转染到THP-1细胞中时，它可以有效阻断脂多糖刺激的THP-1细胞产生白细胞介素-8（p<0.01）。总之，Tec激酶可能介导脂多糖刺激单核细胞中白细胞介素-8的转录。
