Rab5a promotes LPS-induced cytokine expression in macrophages
10.3969/j.issn.1000-484X.2015.03.002
- VernacularTitle:Rab5 a促进LPS活化的巨噬细胞中细胞因子的表达
- Author:
Xiaolin SUN
;
Jiming XIE
;
Xiaole YUN
;
Wei ZHANG
;
Hongbin KANG
;
Yongqing WAN
;
Jingran LIU
;
Pei GONG
;
Shimin ZHAO
;
Yuzhen WANG
- Publication Type:Journal Article
- Keywords:
Rab5a;
LPS;
Macrophage;
iNOS;
TNF-α;
IL-6
- From:
Chinese Journal of Immunology
2015;(3):295-299
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish cell lines stably expressing Rab5a and its the inactive mutant Rab5aN133I,analyze the effect of Rab5a on the expression of cytokines in LPS-stimulated RAW264.7 cells .Methods:RAW264.7 cells were transfected with Rab5a and its inactive mutant vector Rab5a N133I separately,and then screened by G418.Rab5a stable expressing cell lines were identified by Real time-PCR.The growth of the stable cell lines was analyzed by MTT assay.After the stable cell lines were stimulated by LPS for different time periods,the expression of iNOS,TNF-αand IL-6 was detected.Results:Rab5a and Rab5aN133I transfection resulted in elevated Rab5a mRNA expression compared with the control cells ( P<0.05 ).Rab5a overexpression enhanced the proliferation of RAW264.7 cells.However,the proliferation of Rab5aN133I cells was significantly slower than the control cells ( P<0.05).Overexpression of Rab5a promoted LPS-induced production of iNOS,TNF-αand IL-6 in RAW264.7 cells (P<0.01). Conversely,overexpression of Rab5aN133I abolished the stimulating effects of Rab5a.Conclusion: Rab5a promoted LPS-induced expression of iNOS,TNF-αand IL-6 in RAW264.7 macrophages in a GTP-binding ability-dependent manner.
