Mycoplasma MALP-2 induces the expression of heme oxygenase-1 via MAPKs and Nrf2 pathways
10.3760/cma.j.issn.0254-5101.2014.04.007
- VernacularTitle:支原体巨噬细胞活化脂肽2经MAPKs和Nrf2途径诱导人单核细胞血红素氧合酶-1产生
- Author:
Xiaohua MA
;
Xiaoxing YOU
;
Yanhua ZENG
;
Liangzhuan LIU
;
Cuiming ZHU
;
Jun HE
;
Chuanhao JIANG
;
Yimou WU
- Publication Type:Journal Article
- Keywords:
Macrophage-activating lipopeptide-2;
Heme oxygenase-1;
Mitogen-activated protein kinases;
NF-E2-related factor 2
- From:Chinese Journal of Microbiology and Immunology
2014;(4):289-293
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate whether macrophage-activating lipopeptide-2 ( MALP-2) in-duces the expression of heme oxygenase-1 ( HO-1 ) in THP-1 cells and its possible mechanism .Methods Human monocyte cells THP-1 were cultured in vitro and then were incubated with various concentrations (0, 0.01, 0.1, 1.0 or 5.0 ng/ml) of MALP-2 for 16 h, or were stimulated by 5.0 ng/ml MALP-2 for different length of time (0 h, 4 h, 8 h, 12 h, 16 h or 24 h).The expression of HO-1 at mRNA and protein levels were detected by real-time PCR analysis and Western blot assay .The enzyme activity of HO-1 was detected by colorimetric analysis.THP-1 cells were pre-incubated with 30 μmol/L of SB203580, PD98059 and SP600125 for 30 min and then were cultured with 5.0 ng/ml MALP-2 for 16 h to investigate the role of mito-gen-activated protein kinases (MAPKs) signaling pathway in HO-1 production.After incubating THP-1 cells with 5.0 ng/ml MALP-2 for different periods of time, NF-E2-related factor 2 (Nrf2) protein was detected by Western blot assay to study the effects of Nrf2 pathway on MALP-2-induced HO-1 expression.Nrf2 and HO-1 proteins were measured by Western blot assay after transfecting THP-1 cells (1×106/well) with Nrf2 siRNA at a final concentration of 100 nmol/L.Results MALP-2 enhanced the expression of HO-1 at mRNA and protein levels as well as the enzyme activity of HO-1 in THP-1 cells in a concentration-dependent manner.The expression of HO-1 protein induced by MALP-2 was significantly inhibited by 30 μmol/L MAPKs specific inhibitors ( SB203580 , PD98059 and SP600125 ) .MALP-2 induced Nrf2 translocation at a concentration of 5.0 ng/ml.The expression of Nrf2 and HO-1 proteins were significantly decreased in Nrf 2 siRNA-transfected THP-1 cells.Conclusion MAPKs and Nrf2 signaling pathways were involved in the MALP-2 induced HO-1 expression .
