TY - JOUR
T1 - Reactive Oxygen Species-activated p38/ERK 1/2 MAPK Signaling Pathway in the Mycobacterium bovis Bacillus Calmette Guérin (BCG)-induced CCL2 Secretion in Human Monocytic Cell Line THP-1
AU - Méndez-Samperio, Patricia
AU - Pérez, Aline
AU - Alba, Laura
N1 - Funding Information:
This work was supported in part by grant 20101177 from the Secretaría de Investigación y Posgrado (SIP) de el IPN. PMS is an investigator of COFAA, EDI, and SNI.
PY - 2010/11
Y1 - 2010/11
N2 - Background and Aims: CCL2 plays an important role in mycobacterial infection by inducing leukocyte recruitment and activation. Here we assess the role of reactive oxygen species (ROS) in the secretion of the CCL2 and the activation of mitogen-activated protein kinases (MAPKs) by human monocytic cells infected with Mycobacterium bovis bacillus Calmette Guérin (BCG). Methods: CCL2 mRNA and protein expression were measured by reverse transcriptase polymerase chain reaction (RT-PCR), quantitative PCR and ELISA. Kinase phosphorylation was determined by immunoblotting. Results: Treatment of human monocytic cells with M. bovis BCG activated rapid superoxide generation. mRNA expression of CCL2 was increased in M. bovis BCG-infected monocytic cells, and this increase was abrogated by administration of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenyleneiodonium (DPI). Importantly, M. bovis BCG-induced CCL2 protein secretion was also inhibited by the NADPH oxidase inhibitor DPI, the selective inhibitor of NADPH oxidase apocynin, the mitochondrial electron transfer chain subunit I inhibitor rotenone and H2O2 scavenging enzyme catalase, indicating that the inhibition is through the NADPH/ROS pathway. Analysis of downstream signals showed that inhibition of NADPH oxidase inhibited M. bovis BCG-induced phosphorylation of MAPK (extracellular signal-regulated kinase (ERK) 1/2 and p38). Conclusions: These results strongly suggest that NADPH oxidase-derived ROS-mediated activation of p38 and ERK 1/2 is essential for the M. bovis BCG-induced CCL2 production.
AB - Background and Aims: CCL2 plays an important role in mycobacterial infection by inducing leukocyte recruitment and activation. Here we assess the role of reactive oxygen species (ROS) in the secretion of the CCL2 and the activation of mitogen-activated protein kinases (MAPKs) by human monocytic cells infected with Mycobacterium bovis bacillus Calmette Guérin (BCG). Methods: CCL2 mRNA and protein expression were measured by reverse transcriptase polymerase chain reaction (RT-PCR), quantitative PCR and ELISA. Kinase phosphorylation was determined by immunoblotting. Results: Treatment of human monocytic cells with M. bovis BCG activated rapid superoxide generation. mRNA expression of CCL2 was increased in M. bovis BCG-infected monocytic cells, and this increase was abrogated by administration of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenyleneiodonium (DPI). Importantly, M. bovis BCG-induced CCL2 protein secretion was also inhibited by the NADPH oxidase inhibitor DPI, the selective inhibitor of NADPH oxidase apocynin, the mitochondrial electron transfer chain subunit I inhibitor rotenone and H2O2 scavenging enzyme catalase, indicating that the inhibition is through the NADPH/ROS pathway. Analysis of downstream signals showed that inhibition of NADPH oxidase inhibited M. bovis BCG-induced phosphorylation of MAPK (extracellular signal-regulated kinase (ERK) 1/2 and p38). Conclusions: These results strongly suggest that NADPH oxidase-derived ROS-mediated activation of p38 and ERK 1/2 is essential for the M. bovis BCG-induced CCL2 production.
KW - CCL2
KW - Cell signaling
KW - MAPK
KW - Mycobacterium bovis
KW - Reactive oxygen species
UR - http://www.scopus.com/inward/record.url?scp=78650736641&partnerID=8YFLogxK
U2 - 10.1016/j.arcmed.2010.10.009
DO - 10.1016/j.arcmed.2010.10.009
M3 - Artículo
C2 - 21199725
SN - 0188-4409
VL - 41
SP - 579
EP - 585
JO - Archives of Medical Research
JF - Archives of Medical Research
IS - 8
ER -