In vitro effects of epinephrine, norepinephrine, and dobutamine on lipopolysaccharide-stimulated production of tumor necrosis factor-α, interleukin-6, and interleukin-10 in blood from healthy dogs.

OBJECTIVE: To determine the in vitro effects of epinephrine, norepinephrine, and dobutamine on lipopolysaccharide (LPS)-stimulated production of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10) in blood from healthy dogs. SAMPLES: Blood samples from 9 healthy dogs. PROCEDURES: Blood samples were incubated with LPS from Escherichia coli O127:B8 or PBSS (control) for 1 hour. Afterward, the samples were incubated with 10µM epinephrine, norepinephrine, or dobutamine or with saline (0.9% NaCl) solution (control) for 23 hours. Leukocyte viability was assessed by use of trypan-blue exclusion in blood from 2 dogs to ensure cell viability was not altered by the catecholamines. Tumor necrosis factor-α, IL-6, and IL-10 concentrations were measured in the supernatant in duplicate with a canine-specific multiplex bead-based assay. Blood samples from 2 dogs were used to create dose-response curves to evaluate whether the observed cytokine modulation was dependent on catecholamine concentration. RESULTS: Incubation of blood with epinephrine and norepinephrine significantly increased LPS-stimulated production of IL-10, compared with the control. Epinephrine and norepinephrine significantly decreased LPS-stimulated production of TNF-α, compared with the control. Epinephrine and norepinephrine did not significantly alter LPS-stimulated production of IL-6. Dobutamine did not alter catecholamine production. CONCLUSIONS AND CLINICAL RELEVANCE: Epinephrine and norepinephrine, but not dobutamine, had immunomodulatory effects on LPS-stimulated TNF-α and IL-10 production in blood from healthy dogs in this in vitro model of sepsis. Data suggested that dobutamine may have immune system-sparing effects in dogs with sepsis.

Feasibility of hepatic fine needle aspiration as a minimally invasive sampling method for gene expression quantification of pharmacogenetic targets in dogs.

BACKGROUND: Quantifying hepatic gene expression is important for many pharmacogenetic studies. However, this usually requires biopsy (BX), which is invasive. OBJECTIVES: The objectives of this study were to determine the feasibility of using minimally invasive fine needle aspirate (FNA) to quantify hepatic gene expression and to assess expression variability between different sampling sites. METHODS: Biopsy and FNA samples were acquired from central and peripheral locations of the right and left lateral liver lobes of a dog. Relative expression of ABCB1, GSTT1 and CYP3A12 were measured via reverse transcriptase, quantitative PCR. The effect of sampling method, lobe and location within the lobe on gene expression was assessed using a three-way ANOVA. RESULTS: Relative expression of ABCB1 and GSTT1 were not statistically different between sampling methods but CYP3A12 expression was higher in samples collected by BX (p = .013). Lobe sampled affected ABCB1 expression (p = .001) and site within lobe affected ABCB1 (p = .018) and GSTT1 (p = .025) expression. CONCLUSIONS: FNA appears to be a feasible technique for minimally invasive evaluation of hepatic gene expression but results should not be directly compared to biopsy samples. Sampling location impacts expression of some targets; combination of FNAs from multiple sites may reduce variation.