Vlasta K. Zbuzek

Ondansetron exhibits the properties of a local anesthetic.

The purpose of this study was to determine whether ondansetron (OND) has local anesthetic effects.Using a patch-clamp technique, we showed that OND concentration dependently blocked Na channel currents in freshly isolated neurons of rat brains with a 50% inhibition concentration of 12 micro M. The blockade started immediately when OND was applied to the cell body using a fast perfusion system, reached a plateau within 15 s, and recovered to the control level within 30 s after washout of the OND-containing solution. Because this is a known property of local anesthetics, we used the tail-flick technique to verify this effect in vivo in Sprague-Dawley rats (n = 46). OND was injected subcutaneously into the tail at the doses of 0.08, 0.16, and 0.2 mg. The tail-flick latency increased 2 min after OND injection, reaching the plateau within 5 min. This effect was dose-related, lasting from 10 to 25 min. These preliminary data indicate that OND, a selective 5-HT3 receptor antagonist, might serve as a prototype molecule for development of a novel series of local anesthetics. Implications: Ondansetron is a drug used to prevent vomiting, especially in cancer patients after chemotherapy. We found that it also causes numbness when injected under the skin. This new action may contribute to its role in calming the stomach. We studied the effect of ondansetron on the isolated brain cells of live rats. (Anesth Analg 1997;85:1116-21)

Nifedipine-Induced Analgesia After Epidural Injection in Rats

We explored the analgesic effect of epidural nifedipine in male Sprague-Dawley rats. By using an implanted epidural catheter, the rats were given 35 microL of dimethylsulfoxide (DMSO) alone or DMSO containing 2.5, 5, 10, or 20 microM of nifedipine. Analgesia was measured by tailflick (TF) involving spinal reflexes, and by hotplate (HP) requiring an intact central nervous system. The latencies were recorded up to 120 min after the injection. The cutoff time of the noxious stimuli was 20 s in the TF and 60 s in the HP to prevent tissue damage. The TF technique revealed a significant difference from the control at doses of 5, 10, and 20 microM with no difference among the groups. Maximum latencies (cutoff time) lasted for 15, 30, and 40 min at doses of 5, 10, and 20 microM, respectively. The HP technique disclosed a dual effect: a significant decrease at the dose of 2.5 microM, no effect at 5 microM, and an increase at 10 and 20 microM. However, the median latency did not reach the cutoff time. We conclude that nifedipine, given epidurally, possesses antinociceptive properties at the dose of 5 microM and higher, detected better by the TF than HP. Our data suggest that the antinociceptive effect of nifedipine, at the studied doses, is more prominent at the spinal than the supraspinal level.

The effect of aging on vasopressin system in Fisher 344 rats.

Vasopressin (VP) was measured by RIA in the plasma, neurohypophysis (NH) and hypothalamus (HT) of young (2 months), adult (12 months) and old (30 months) male Fisher 344 rats, ten of each age. Plasma VP concentration was significantly lower in old compared to adult and to young rats. VP content in the NH expressed per mg weight was similar in all three groups, while the hypothalamic VP content was decreased in the aged rats. This suggests that reduced synthesis and release and/or increased degradation of VP occurs in aged rats. These data are in an agreement with our previous results obtained with Sprague-Dawley rats indicating that these differences are not strain-related. In a pilot experiment, we further studied VP release from isolated perifused NH of young and old rats. Two glands of each age were simultaneously perifused in individual microchambers with medium TC 199 and subsequently stimulated by Lockes medium containing 56 mM K+. Both, the initial and the basal VP release from the NH of the old rats, as well as the response to high K+, were about a half that of the young rats. Thus, a decreased VP release may contribute to the findings of lower plasma VP concentration in aged rats.

Ondansetron blocks nifedipine-induced analgesia in rats

The serotonergic system is involved in pain transmission and the 5-hydroxytryptamine (5-HT3) receptor subtype mediates some of these effects at the spinal level. Therefore, we explored the effects of the serotonergic system on nifedipine-induced analgesia by using the 5-HT3 receptor antagonist ondansetron. Male Sprague-Dawley rats were pretreated with ondansetron (1 mg/kg intraperitoneally) or normal saline. After 15 min, rats received injections of nifedipine (15 mg/kg intraperitoneally) or dimethylsulfoxide (DMSO), solvent for nifedipine, as a control. Nociception was assessed by tail-flick method. Rats treated with nifedipine alone had an increase in tail-flick latency of 122%, as measured by the area under the curve, compared to rats treated with DMSO alone. Pretreatment with ondansetron, however, completely blocked the analgesic effect of nifedipine, with tail-flick latency remaining at baseline throughout the measurement period. These results indicate that the 5-HT (3) receptor plays an important role in the analgesic response to nifedipine and that medications that block this receptor may decrease the analgesic effectiveness of this type of therapy. (Anesth Analg 1996;82:498-500)

Nicotene-induced analgesia in rats: The role of calcium and the diversity of responders and nonresponders

Following a single dose of nicotine, (NIC, 1 mg/kg s.c.), 60% of tested rats revealed significant antinociception as measured by the tail-flick (TF) test, and were classified as responders, with those in which TF latencies did not change, nonresponders. The following experiments were carried out one week later. In nonresponders, pretreatment with ethylenediaminetetraacetic acid (EDTA, 250 microM/kg s.c. four times every 15 min) followed by 1 mg NIC, produced significant analgesia in 50% of rats, to the same magnitude as did nicotine alone (1 mg) in responders. The other 50% of rats which failed to respond to EDTA pretreatment, all revealed similar analgesia following the higher dose of NIC (1.5 mg/kg s.c.), with similar side effects, as generally observed in responders. In responders, pretreatment with CaCl2 (1.5 mM/kg s.c.) completely abolished NIC (1 mg/kg s.c.)--induced analgesia in all rats. Our data provide stronger evidence and a further verification that EDTA potentiates, whereas CaCl2 completely abolishes, nicotine-induced analgesia in rats; supporting our hypothesis of the involvement of calcium ions in this effect.

Vasopressin Release from Individually Superfused Neurohypophyses Decreases in Aged Rats

We studied the effect of aging on vasopressin (VP) release from isolated neurohypophyses (NH) individually superfused with synthetic medium TC 199 with Hanks salts. The superfusion technique is described in detail. Male Fisher 344 rats 2 months (young, n = 17), 12 months (adult, n = 14) and 30 months (old, n = 17) of age were used. VP was measured by radioimmunoassay (RIA). The age-related VP release was analyzed as follows: initial traumatic release, basal release, VP release evoked by electrical stimulation (10-second trains every 20 s of matched biphasic stimuli, 0.2 ms width, 8 mA, 30 Hz), by chemical stimulation (56 mM K+) and total release for the entire superfusion period. When VP release was expressed per milligram NH, it was significantly lower under all conditions in the old rats than in the young ones. In the adult rats, traumatic, basal and total release values were similar to those of the young rats, whereas their responses to chemical and electrical stimulation were similar to those of old rats. Residual VP content expressed per whole NH was significantly higher in the old and adult rats, reflecting a larger glandular size, but when expressed per milligram NH tissue, it was low in the old rats. The percent of VP released during the entire superfusion period relative to the residual VP content was significantly lower in the old than in the young and adult rats. The magnitude of the maximal VP release exceeding the basal release, in response to electrical and chemical stimulation, was similar in young and old rats. However, in the adult rats it was significantly lower than in both the young and old following chemical stimulation. A significantly larger number of old and adult NHs exhibited a more delayed response to chemical, but not to electrical stimulation, than did the young NHs. These data demonstrate an age-related decrease in VP release in Fisher 344 rats. Since the traumatic and basal VP release in the adult rats is similar to the release in the young rats, while the stimulated release in the adult rats resembles the response of the old animals, the results suggest that an impairment of stimulated VP release occurs at an earlier stage of the aging process than does an impairment of spontaneous (traumatic and basal) release.

GABAA ANTAGONIST AND NICOTINE-INDUCED ANTINOCICEPTION

Rats, pretreated with saline or GABA(A) antagonist bicuculline (BIC) at the doses of 2, 4, 6, 10 and 20 mg/kg, were injected with nicotine (NIC, 1 mg/kg) 30 min later. Tail-flick (TF) latencies were measured before (baseline) and three times at 10 min interval after pretreated compounds, continued every 10 min up to 1 hr after NIC injection. In all groups, median TF latencies did not change from the baseline for the first 30 min after the pretreatment. Following NIC alone (control) and in the group pretreated with 2 mg/kg BIC, 60% rats reached ceiling TF latencies (20 sec) lasting for 10 min. In groups with higher BIC doses (4 to 10 mg/kg), median TF latencies were in the range of 5-7 sec with 30% rats reaching the ceiling TF latencies. Following 20 mg/kg BIC, one out of five rats reached 20 sec; the median was in the range of 4-5 sec. Significantly lower responses were observed following 4 mg BIC and higher doses with no difference among the groups. In conclusion, our novel data show that BIC alone, injected systemically, does not possess any effect on the thermal nociceptive transmission as measured by the tail flick test. However, pretreatment with BIC partially prevents NIC-induced antinociception, in a non dose related manner. This suggests that GABA(A) receptors may, at least in part, contribute to the complex mechanisms involved in NIC-induced antinociception.

The effects of serotonin biosynthesis inhibition on nicotine and nifedipine-induced analgesia in rats.

The calcium channel blocker nifedipine has analgesic properties that are enhanced by nicotine.Although it is not known how this analgesic state might affect the awareness of anginal pain and impending myocardial infraction, recent studies have shown an increased mortality associated with the use of large doses of nifedipine. Because both nifedipine- and nicotine-induced analgesia involve serotonergic mechanisms, we studied the effects of the serotonin biosynthesis inhibitor parachlorophenylalanine (pCPA) on nifedipine- and nicotine-induced analgesia. Nociception was assessed by tail-flick method. Rats pretreated with pCPA (300 mg/kg intraperitoneally [IP]) followed by either nifedipine (15 mg/kg IP) or nicotine (1 mg/kg subctaneously) had a increase in tail-flick latency of 41% (P = 0.09) and 50% (P = 0.05), respectively, compared with animals that did not receive pCPA. Additionally, rats pretreated with pCPA followed by a combination of nicotine and nifedipine doubled their tail-flick latency (P = 0.0001) compared with animals that were not treated with pCPA. These data further support the involvement of the serotonergic system in both nifedipine- and nicotine-induced analgesia and suggest that drugs that affect serotonin levels, including tricyclic antidepressants and serotonin-specific reuptake inhibitors, may also affect the analgesia induced by nifedipine and nicotine. Implications: This study examines the effect of serotonin depletion on nicotine- and nifedipine-induced analgesia. Nifidipine is a calcium channel blocker used to treat high blood pressure. It also has pain-relieving properties that are enhanced by nicotine. Because both nifedipine- and nicotine-induced analgesia involve the neurotransmitter serotonin, it is important to know how changes in serotonin concentration might affect both nicotine- and nifedipine-induced analgesia. This study not only supports the involvement of the serotonergic system in both nifidipine- and nicotine-induced analgesia, but also suggests that drugs that affect serotonin levels may also affect analgesia induced by nifidipine and nicotine. (Anesth Analg 1998;87:1109-12)

Hypotension does not alter the antinociceptive effect of nifedipine

We explored the relationship between antinociceptive and hypotensive effects of nifedipine (NIF) injected intraperitoneally ( ip, 15 mg/kg) and epidurally (epi, 20 microM), as compared to verapamil (VER, 10 mg/kg ip) and nitroglycerin (NTG, 0.1 and 0.15 mg/kg ip). The systolic blood pressure (BP) and tail-flick (TF) latencies were measured simultaneously every 10 min for 2 hours and individual values of both measurements were correlated. The highest antinociceptive as well as hypotensive effects were both measured in the group receiving NIF epi., with the correlation coefficient r2=0.2878. Injected ip., NIF revealed similar antinociceptive effect, whereas the other studied drugs were not effective. As to the degree of hypotensive activity, NIF epi was followed by VER, NTG 0.1, NIF ip. and NTG 0.15. No significant correlation was found between BP and TF latencies in any group receiving the drugs. We concluded that the antinociceptive response, measured by the tail-flick technique, is independent of the hypotensive activity of the studied drugs, including NIF.

Effect of Chronic Nicotine Treatment and its Withdrawal on the Vasopressinergic System in Rats

We have studied the effect of chronic nicotine treatment and its withdrawal on the hypothalamo‐neurohypophyseal Vasopressinergic system in male Sprague‐Dawley rats. They were subcutaneously infused with low and high doses of nicotine, free base (0.6 and 6.0 mg/kg/day, respectively) for 28 days, via Alzet osmotic pumps. The studies were carried out immediately after the period of infusion and 1, 7, 14 and 28 (the latter in high dose only) days later. Basal, high K+‐stimulated and total vasopressin release from the superfused neural lobes, the residual vasopressin content in the neural lobes, and hypothalamus and plasma vasopressin concentration were measured by radioimmunoassay.