Regulation of nitric oxide production from macrophages by lipopolysaccharide and catecholamines☆
Section snippets
Experimental procedures
Materials. Mouse Raw264.7 macrophage cell line was obtained from American Type Culture Collection (ATCC, Manassas, VA). Raw264.7 cell line was maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum, penicillin (50 U/ml), and streptomycin (50 μg/ml). LPS (Escherichia coli serotype 0111:B4), epinephrine, norepinephrine, and dopamine were purchased from Sigma (St. Louis, MO).
Effect of LPS and catecholamines on NO production in macrophages. Two milliliters of RAW264.7 murine
Results
Effect of catecholamines and LPS on nitrite production from macrophages. Two milliliters of RAW264.7 murine macrophage cells (1.8×106/ml) were cultured in 12-well tissue culture plates with medium or LPS at a concentration ranging from 0.5 to 10 ng/ml for 24 h. The nitrite production from macrophages treated with LPS was increased in a dose-dependent manner. LPS at concentrations of 5–10 ng/ml significantly (P<0.05) increased nitrite production. Epinephrine (Epi), norepinephrine (Nor), or dopamine
Discussion
Catecholamines are elaborated in stress responses to mediate vasoconstriction and elevate systemic vascular resistance and blood pressure. Sympathetic nervous system activation together with microbial infection results in a synergistic interaction that could modulate inflammatory processes [26]. In septic shock, NO has been shown to be involved in vascular collapse, which is a major contributor to mortality. Thus, it is important to understand the interaction between catecholamines and LPS with
Acknowledgements
This work was supported by NIH Grant HL63070, the Ruth R. Harris Endowment, and RDC and CVRI of ETSU.
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Part of this work was presented in preliminary form at the Immunology 2000 AAI/CIS Joint Annual Meeting, Seattle, WA, May 12–16, 2000.