The regulatory effect of ERK1/2 signal pathway on production of TNFα induced by LPS in mice Kupffer cells
- Author:
Yu ZHANG
1
;
Jianxin JIANG
;
Shanhe JI
;
You'an SHAN
;
Peifang ZHU
;
Jihong ZHOU
Author Information
1. Research Institute of Surgery Daping Hospital Third Military Medical University
- From:
Chinese Journal of Traumatology
2001;4(3):139-142
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To study the rule of ERK1/2 activity and regulative effect of ERK1/2 pathway on the production of pro-inflammatory cytokine TNFα in mice Kupffer cells (mKC) induced by LPS, and to exploring novel methods to prevent and treat clinical patients of endotoxemia. Methods: Immunoprecipitate kinase assay and Western blotting analysis were used to detect the phosphorylated ERK1/2 kinase activity in mKC stimulated by LPS, and ELISA was used to study the effect of ERK1/2 signaling cascade on LPS-induced TNFα production in mKC. Results: In mKC, LPS treatment resulted in transient and rapid increase of kinase activity of ERK1/2 that phosphorylated their specific substrate ELK-1, with maximal value at 30 minutes and a return near to baseline within 2 hours, and LPS-induced ERK1/2 activity from LPS concentration of 10 pg/ml to the top activity at 100 ng/ml. No activity was observed in unstimulated mKC. Inhibition of the ERK1/2 pathway using the specific ERK1/2 signal pathway inhibitor PD98059 caused a marked and concentration-dependent reduction of TNFα production. Conclusions: The results show that LPS can markedly activate ERK1/2 pathway in mKC. PD98059 causes a significant and concentration-dependent reduction of TNFα production. ERK1/2 may be a novel target to treat clinical patient of endotoxemia.
