Rab5a promotes expression of pro-inflammatory cytokines and type Ⅰ IFN in CpG induced macrophages
10.3969/j.issn.1000-484X.2016.02.004
- VernacularTitle:Rab5a促进CpG诱导的巨噬细胞中炎性细胞因子和Ⅰ型干扰素的表达
- Author:
Junna CHEN
;
Xiaolin SUN
;
Shichao DONG
;
Kai ZOU
;
Huan LIU
;
Yan WANG
;
Xue WANG
;
Fang LIU
;
Jiming XIE
;
Yuzhen WANG
- Publication Type:Journal Article
- Keywords:
Rab5a;
CpG;
TLR9;
TNF-α;
IL-1β;
IFN-β
- From:
Chinese Journal of Immunology
2016;(2):165-168
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Using the macrophage cell lines RAW264.7 stably expressing Rab5a and its dominant negative mutant Rab5aN133I as models to analyze the effect and the mechanism of Rab 5a,Rab5aN133I on CpG-induced production of pro-inflammatory cytokines and type Ⅰ IFN.Methods: The eukaryotic expression vectors of Rab5a and Rab5aN133I were transfected into RAW264.7 cells by liposome,and screened with G418.The G418-resistant colonies were obtained and amplified.The transformed cell lines were i-dentified by RT-PCR,Real time-PCR and Western blot.The production of cytokines were measured after transformed cell lines of Rab5a and Rab5aN133I was stimulation with CpG for 8 h.Results: Rab5a expression in transfected cells was significantly higher than the control cell group (P<0.05).Overexpression of Rab5a significantly promoted the production of TNF -α,IL1-β(P<0.01) and IFN-β( P<0.05) in CpG stimulated RAW264.7.The production of cytokines was restored in Rab 5aN133I transfected cell line.Conclusion:Rab 5a promotes CpG-induced pro-inflammatory cytokines and typeⅠIFN in macrophages,it may be act as a positive regulator in TLR9 signaling pathway.
