Endogenous peripheral antinociception in early inflammation is not limited by the number of opioid-containing leukocytes but by opioid receptor expression

Abstract 

Endogenous inhibition of inflammatory pain is mediated by leukocytes that secrete opioid peptides upon exposure to stress (cold water swim stress, CWS) or after local injection of corticotropin releasing factor (CRF). Since in early inflammation few opioid-containing leukocytes are detected and since peripheral opioid-mediated antinociception is low we examined whether antinociception could be augmented by increased recruitment of opioid-containing polymorphonuclear cells (PMN). Rats were intraplantarly (i.pl.) injected with Freund's complete adjuvant (FCA) and with the PMN-recruiting chemokine macrophage inflammatory protein-2 (MIP-2, 1–10 μg; control: saline) for 2 h. Intraplantar leukocytes were quantified by flow cytometry. Paw pressure threshold (PPT) was determined before and after exposure to CWS, i.pl. injection of CRF and opioid peptides. Opioid receptors (OR) were measured by binding studies in dorsal root ganglia (DRG) and by immunohistochemistry in the paw. Our studies showed that (i) MIP-2 injection dose-dependently augmented recruitment of PMN and opioid-containing leukocytes (5-fold increase in cells/paw, P<0.05), (ii) PPT was not different between groups at baseline and after CWS or CRF (maximum MPE: 20±2.3–29±7.2%, P<0.05), (iii) injection of opioid peptides dose-dependently increased the PPT (P<0.05, maximum MPE: and 18±2.6–21±3.6%), (iv) MOR (μ OR, MOP) binding sites in the ipsilateral DRG were unchanged (24±2–22±1.2fmol/mg protein, P>0.05, ANOVA) and (v) the number of MOR and DOR (δ OR, DOP) stained nerve fibers in peripheral tissue were unaltered (both P>0.05, t-test). In summary, antinociception during early inflammation is apparently not limited by the number of opioid-containing leukocytes but by OR availability.

Keywords:  Freund's complete adjuvant, Chemokine, β-endorphin, PMN, Opioid receptors

Abbreviations:  mAb, monoclonal antibody, BL, baseline, CRF, corticotropin releasing factor, CWS, cold water swim stress, CyC, Cy chrome, DAMGO, [D-Ala²,N-MePhe⁴,Gly⁵-ol]enkephalin, DOR, (=DOP) δ-opioid receptor, DPDPE, ([D-Pen²,D-Pen⁵]-enkephalin, DRG, dorsal root ganglia, END, β-endorphin, ENK, Met-enkephalin, FACS, fluorescence activated cell staining, FCA, Freund‘s complete adjuvant, FITC, fluorescein isothiocyanate, MIP-2, macrophage inflammatory protein-2, MOR, (=MOP) μ-opioid receptor, MPE, maximum possible effect, NLX, naloxone, NTI, naltrindole, PE, phytoerythrin, PMN, polymorphonuclear cells, PPT, paw pressure threshold