ABSTRACT
BACKGROUND: We have previously shown that children with mild asthma have a modest improvement in their pulmonary function tests after aerosolized furosemide. The mechanism of action is not known. The observation that furosemide possesses a similar profile of protection as sodium cromoglycate and nedocromil sodium suggests that furosemide may inhibit mediator production and release. OBJECTIVE: We studied the in vitro effects of furosemide on cytokine release from normal human peripheral blood mononuclear cells (PBMC) induced by E. coli lipopolysaccharide (LPS). METHODS: Peripheral blood mononuclear cells were isolated by density gradient centrifugation, stimulated with LPS and incubated at 37 degrees C with varying concentrations of furosemide, hydrocortisone, sodium cromoglycate, and nedocromil sodium for 24 hours. Supernatants were extracted and study for levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and interleukin-8 (IL-8). Intracellular IL-6 and TNF-alpha concentrations were also measured by cell cytometry. Cell viability was examined using XTT cell proliferation test and-measuring the release of lactate dehydrogenase (LDH). RESULTS: There was a significant reduction in levels of TNF-alpha and IL-6 at a furosemide concentration of 0.5 x 10(-2) M and a reduction in IL-8 levels at 10(-2) M. This inhibition was comparable to that found with equivalent molar concentrations of hydrocortisone. These findings were also confirmed with measurements of intracellular IL-6 and TNF-alpha by cell cytometry. High concentration of furosemide at 10(-2) M caused significant cellular cytotoxicity. CONCLUSION: These data suggest that furosemide may exhibit an anti-inflammatory effect. Specifically, the addition of furosemide resulted in decreased production of cytokines. This effect may be due to an immunosuppressive activity on monocytes as well as a direct cytotoxic effect at high furosemide concentrations.
Subject(s)
Furosemide/immunology , Immunosuppressive Agents/pharmacology , Leukocytes, Mononuclear/drug effects , Anti-Asthmatic Agents/pharmacology , Cell Division/drug effects , Cell Division/immunology , Cell Survival/drug effects , Cytokines/drug effects , Cytokines/metabolism , Diuretics/immunology , Diuretics/toxicity , Flow Cytometry , Furosemide/toxicity , Humans , Interleukin-6/metabolism , Interleukin-8/metabolism , Intracellular Fluid/chemistry , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/metabolism , Staining and Labeling , Tumor Necrosis Factor-alpha/drug effects , Tumor Necrosis Factor-alpha/metabolismABSTRACT
The effects of prenatal cocaine exposure on the levels of carotid body dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were investigated in 5-day-old rat pups exposed to normoxic and hypoxic conditions. Timed-pregnant Sprague-Dawley rats were injected b.i.d. with either cocaine HCl (30 mg/kg) or isotonic saline (1 ml/kg) from gestational days 7-21. On the fifth postnatal day, pups were subjected to either 20 min of 0.21 or 0.08 fractional inspired oxygen (FlO2). Under a strictly timed protocol, both carotid bodies were removed from each pup, placed in an antioxidant solution to prevent DA breakdown, and subsequently analyzed via HPLC with electrochemical detection to determine carotid body DA and DOPAC content. Two-way ANOVA revealed decreases in DA in cocaine-exposed pups. No HVA was detectable in any of the samples. The 0.08 FlO2 condition decreased DA compared to 0.21 FlO2. The additive consequences of DA depletion resulting from the combination of prenatal cocaine and postnatal hypoxia decreased carotid body DA to 14% of control levels, with several animals exhibiting DA content below detection limits. Considering the role of the carotid body in the ventilatory response to hypoxia, these data suggest that prenatal cocaine exposure may adversely affect the normal chemoreceptive function of the carotid body.
