The Eighteenth Days of Neuropsychopharmacology Posters Posters Impairment in pain perception in adult rats treated with N-(-2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) as neonates Beata Adamus-Sitkiewicz, Eva Kõrössy, Kamila Bojanek, Marta Adwent, Micha³ Ba³asz, Ma³gorzata Kniaœ, Ryszard Szkilnik Department of Pharmacology, Medical University of Silesia, Jordana 39, PL 41-808 Zabrze, Poland To examine the impact of a central noradrenergic le- paracetamol was also greatly diminished in the DSP-4 sion on antinociceptive effects of morphine, paraceta- group and significant in all tested intervals. Nefopam mol and nefopam, we compared intact male rats with produced only slight analgesia in both tested groups. rats in which noradrenergic nerve terminals were In biochemical studies we showed that in DSP-4 largely destroyed with the neurotoxin DSP-4 [N-(2-chlo- treated rats there was a marked decrease in NA level roethyl)-N-ethyl-2-bromobenzylamine; 50 mg/kg, sc ×2] in the prefrontal cortex (to 10.4%, p < 0.005), the shortly after birth, on the 1st and 3rd day of postnatal thalamus with the hypothalamus (to 54.4%, p < 0.005) life. When the rats attained 10 weeks of age, painful and the spinal cord (to 12.3%, p < 0.005) in compari- reactions were assessed by means of the tail immer- son to the control group. Conversely, in the cerebel- sion test (thermal stimulus) and the paw pressure test lum and brain stem of rats with DSP-4 lesions there (mechanical stimulus). In addition, monoamine levels was a significant increase in the NA content vs. con- in some parts of the brain were estimated using the trol (to 171.2% and 123.5% of NA, respectively, with high pressure liquid chromatography with electro- p < 0.005 and p < 0.05, respectively). In the striatum chemical detection (HPLC/ED) method. In the tail we did not observe any changes in NA level between immersion test, we showed that there were no differ- the examined groups. The levels of 5-HT and its me- ences in the antinociceptive effect evoked by mor- tabolite 5-HIAA were also not altered by DSP-4 treat- phine (5.0 mg/kg, sc) and paracetamol (100 mg/kg, ment in all tested structures with the exception of the ip) between control and DSP-4 rats. Nefopam spinal cord (approx. 40% decrease) and the level of (20 mg/kg, ip) elicited only slight analgesia in control DOPAC (also 40% reduction). In conclusion, ob- rats (~17%), and this effect was not observed in the tained results showed that neonatal DSP-4 treatment DSP-4 treated group; differences were statistically alters the antinociceptive effects of tested drugs (each significant at 90 and 120 min of this test. In the paw of them with a different mechanism of action). These pressure test we demonstrated that morphine pro- data lead to the proposal that perhaps there is a need duced lower analgesia in DSP-4 rats in comparison to to adjust the doses of analgesics applied to patients the control, and the effect was significant at 60, 90 with noradrenergic system dysfunction (e.g., depres- and 120 min of the test. The antinociceptive effect of sion and/or anxiety disorders). Pharmacological Reports, 2009, 61, 369379 369 Inflammatory and visceral pain perception in rats lesioned with DSP-4 as neonates Beata Adamus-Sitkiewicz, Eva Kõrössy, Kamila Bojanek, Marta Adwent, Micha³ Ba³asz, Urszula Miko³ajun, Rafa³ Muchacki, Ryszard Szkilnik Department of Pharmacology, Medical University of Silesia, Jordana 39, PL 41-808 Zabrze, Poland We investigated the reactivity of N-(2-chloroethyl)- Paracetamol elicited lower analgesia in control in N-ethyl-2-bromobenzylamine lesion (DSP-4; 50 mg/kg, comparison to DSP-4 rats; the effect was significant sc × 2, P1 and P3) on the inflammatory and visceral at 20–30, 30–40 and 40–50 min intervals of observa- pain perception after morphine, paracetamol and ne- tion. Nefopam was without effect in this regard. In fopam administration. When rats attained 10 weeks of biochemical assays, equally high levels of 5-HTP in age, painful reactions were assessed by means of the the prefrontal cortex, thalamus with hypothalamus formalin test (inflammatory stimulus) and the writh- and brainstem were observed when comparing control ing test (chemical stimulus). Furthermore, accumula- and DSP-4 lesioned animals (after 0.9% saline). Mor- tion of L-dihydroxyphenylalanine (L-DOPA) and phine significantly increased the 5-HTP level only in 5-hydroxytryptamine (5-HTP) after administration of the prefrontal cortex of control rats. Paracetamol ele- the aromatic amino acids inhibitor – hydroxybenzyl- vated 5-HTP content in the DSP-4 group in the pre- hydrazine (NSD-1015) 100 mg/kg, ip in some part of frontal cortex but diminished the 5-HTP levels in the thalamus with hypothalamus; at the same time no ef- the brain was examined using high pressure liquid fect was observed in control animals (in all tested chromatography with the electrochemical detection brain parts). Conversely, nefopam decreased 5-HTP (HPLC/ED) method. Thirty minutes after a morphine content in the prefrontal cortex and thalamus with hy- (5.0 mg/kg, sc) challenge, rats were subcutaneously pothalamus of DSP-4 rats but no effect was noted in injected into the plantar surface of the right hind paw the brain stem. Nefopam did not cause accumulation with 50 µl 5% formalin solution. Both groups showed of 5-HTP in control rats. Equally high levels of the typical biphasic nocifensive response curve lasting L-DOPA in all examined parts of the brain were also 60 min of testing but DSP-4 lesioned rats scored more noted when comparing control and DSP-4 lesioned points (spending more time licking/biting the injected animals after saline administration. Morphine did not hind paw) in the first and second phase as well as the affect L-DOPA levels in any tested structures of either interphase period of the formalin test than the control experimental group of rats. Nefopam reduced L-DOPA group. After paracetamol (100 mg/kg, ip) and nefo- only in the brainstem of DSP-4 treated animals in com- pam (20 mg/kg, ip) administration, typical biphasic parison to control (after nefopam administration) and nocifensive response curve were also observed; how- DSP-4 after saline injection. The results of the present ever, no differences between control and DSP-4 study demonstrated that noradrenergic system dysfunc- treated rats were noticed. Injections of morphine tion produced by neonatal DSP-4 treatment modified evoked similar antinociception in the visceral pain the antinociceptive effects of the examined analgesics. model in both tested groups (control and DSP-4). 370 Pharmacological Reports, 2009, 61, 369379 The Eighteenth Days of Neuropsychopharmacology Posters Influence of imipramine, moclobemide and fluoxetine on pro-inflammatory (TNF-a, IL-1b) and anti-inflammatory (IL-10) cytokines in lipopolysaccharide-stimulated primary rat mixed glial cell cultures Anna M. Bielecka, Anna Pude³ko, Ewa Obuchowicz Department of Pharmacology, Medical University of Silesia, Medyków 18, PL 40-752, Poland; e-mail: [email protected] It is suggested that glial activation and neuroinflamma- The results have shown that imipramine, moclobe- tory processes play an important role in the pathogenesis mide and fluoxetine decrease TNF-a and IL-1b con- of psychiatric and neurodegenerative diseases. Activated centrations in the culture medium but they have no glial cells secrete various cytokines that influence neuro- significant influence on IL-10 levels. Moclobemide transmission, HPA axis activity and neuronal plasticity, and fluoxetine reduced TNF-a release at concentra- and may contribute to neuronal cell death. tions from 108 to10 µM. Imipramine produced this The anti-inflammatory effects of antidepressants effect at concentrations from 108 to 100 µM. Inhibi- with different influences on monoaminergic systems tion of IL-1b release was observed when moclobe- (imipramine, moclobemide, fluoxetine) were investi- mide or fluoxetine was given at concentrations from gated using 13–14 day primary rat mixed glial cul- 106 to10 µM. Imipramine induced the same effect at tures stimulated by lipopolysaccharide (LPS). Cell concentrations from106 to 100 µM. Levels of IL-10 cultures were prepared from cerebral hemispheres of did not changed significantly after administration of one-day old newborn Wistar rats. Levels of TNF-a, the studied drugs at concentrations from 106 to 100 IL-1b, and IL-10 were evaluated in culture medium (or to 10 µM for fluoxetine). with ELISA kits. The cultures were stained with Ric- Our results support the observation that antidepres- inus communis agglutinin-1 and an antibody against sants have anti-inflammatory effects in central nervous GFAP. The strongest stimulation of TNF-a release was system because they affect the balance between pro-and observed after 6-h incubation with 1 µg/ml of LPS, and anti-inflammatory cytokines (IL-1b, TNF-a/IL-10) in for IL-1b and IL-10 after 48-h incubation with 2 µg/ml of primary mixed glial cell cultures. LPS. Antidepressants were used in concentrations rang- 8 ing from 10 to 100 µM. Fluoxetine was only applied at Acknowledgment: a concentration up to 10 µM because the higher concen- This work was supported by Grant N 401 130 31/2871 from the tration was cytotoxic as determined by MTT and the Try- Ministry of Science of Higher Education. pan Blue exclusion method. Influence of baclofen and group I mGluR ligands on activity of rats in the Porsolt test Halina Car, Ró¿a J. Wiœniewska Department of Pharmacology, Medical University of Bia³ystok, Mickiewicza 2c, PL 15-222 Bia³ystok, Poland, e-mail [email protected] Depression is associated with disturbances in trans- brain. Data showing the antidepressant activity of missions in the glutamatergic and GABAergic sys- ligands for the metabotropic glutamate receptors tems, producing a misbalance of proper activity in the group I (I mGluRs) have been published.