Células cebadas en pulmón y nervio periférico en la intoxicación crónica con karwinskia humboldtiana en rata wistar: estudios histológico e histoquímico / Lung and peripheral nerve mast cells in chronic intoxication with karwinskia humboldtiana in wistar rat: histological and histochemical studies

Karwinskia humboldtiana (Kh) es un arbusto venenoso responsable de numerosos casos de intoxicación accidental en humanos. En estudios previos en nuestro laboratorio reportamos un incremento de células cebadas en nervio periférico (NP) durante la intoxicación con Kh, este hallazgo no ha sido reportado previamente en otros órganos durante esta intoxicación por lo que en el presente estudio buscamos la presencia de estas células en otros órganos, además de distinguir subpoblaciones de células cebadas mediante reacciones histoquímicas para la identificación de los gránulos de secreción. El objetivo de este estudio fue evaluar la presencia de células cebadas en órganos distintos al NP y diferenciar histoquímicamente la composición de sus gránulos. Se utilizaron 32 ratas Wistar, se dividieron en cuatro grupos (n= 8) en donde 5 ratas de cada grupo fueron intoxicadas y 3 fueron control no intoxicadas. A las ratas intoxicadas se les administraron por vía oral 3,5 g/kg del fruto seco y molido de Kh fraccionados en 5 dosis de 1,5; 0,5, 0,5; 0,5 y 0,5 g/kg los días 0, 3, 7, 10 y 14 respectivamente. Las ratas control solo recibieron agua. Cada grupo fue sacrificado a diferentes tiempos según la evolución de la parálisis. Se obtuvieron muestras de Hígado, Riñón, Pulmón y SNP, se procesaron hasta obtener bloques de parafina, se obtuvieron cortes y se tiñeron con azul de toluidina, PAS, Azul alciano/PAS y Azul alciano/Safranina. Se identificó la presencia de células cebadas en NP y pulmón con la tinción de azul de toluidina y se realizo un estudio morfométrico observando un incremento progresivo del número de células cebadas por grupo así como variaciones histoquímicas en sus gránulos en cada etapa y órgano analizado, lo que sugiere la participación de las células cebadas y sus secreciones en cada una de las etapas de la intoxicación crónica con el fruto maduro de Kh.

Karwinskia humboldtiana (Kh) is a poisonous shrub causing a number of accidental intoxications in humans. In previous studies in our laboratory, we reported an increased number of mast cells present in peripheral nerve of Kh intoxicated rats. This finding has not been reported in other organs of intoxicated animals. For this reason, in the present study we searched for mast cells in several organs, identifying mast cell subpopulations on the basis of different histochemical reactivity of their secretory granules. Thus the objective of the present study was to evaluate the presence of mast cells in organs other than peripheral nerve and, to distinguish mast cells by their granule content, applying histochemical reactions. 32 Wistar rats were divided into 4 groups (n=8). For each group, 5 rats were intoxicated with Kh and 3 received water only as a control.Intoxicated rats received 3.5 g/ Kg body weight of dry powder of Kh fruits, fractionated in 5 doses as follows 1.5, 0.5, 0.5, 0.5, 05 on days 0, 3,7,10,14 respectively. Control rats received water only. Each group was killed at different times during paralysis evolution. Samples of liver, kidney, lung and brain, were obtained and processed by routine technique until paraffin embedding. Sections were obtained and stained with toluidine blue, PAS, alcian blue/PAS and alcian blue/safranin. Mast cells infiltrates were observed in peripheral nerve and lung. Mast cells were counted. An increasing number of mast cells were recorded as well as variations in the histochemical pattern of their granules for each organ. These findings suggest a role for mast cells and their secretions in the intoxication with mature fruit of Kh.

Amphetamine-type central nervous system stimulants release norepinephrine more potently than they release dopamine and serotonin.

A large body of evidence supports the hypothesis that mesolimbic dopamine (DA) mediates, in animal models, the reinforcing effects of central nervous system stimulants such as cocaine and amphetamine. The role DA plays in mediating amphetamine-type subjective effects of stimulants in humans remains to be established. Both amphetamine and cocaine increase norepinephrine (NE) via stimulation of release and inhibition of reuptake, respectively. If increases in NE mediate amphetamine-type subjective effects of stimulants in humans, then one would predict that stimulant medications that produce amphetamine-type subjective effects in humans should share the ability to increase NE. To test this hypothesis, we determined, using in vitro methods, the neurochemical mechanism of action of amphetamine, 3,4-methylenedioxymethamphetamine (MDMA), (+)-methamphetamine, ephedrine, phentermine, and aminorex. As expected, their rank order of potency for DA release was similar to their rank order of potency in published self-administration studies. Interestingly, the results demonstrated that the most potent effect of these stimulants is to release NE. Importantly, the oral dose of these stimulants, which produce amphetamine-type subjective effects in humans, correlated with the their potency in releasing NE, not DA, and did not decrease plasma prolactin, an effect mediated by DA release. These results suggest that NE may contribute to the amphetamine-type subjective effects of stimulants in humans.

Opioid peptide receptor studies. 12. Buprenorphine is a potent and selective mu/kappa antagonist in the [35S]-GTP-gamma-S functional binding assay.

We utilized the [(35)S]-GTP-gamma-S functional binding assay to determine the selectivity of opioid receptor agonists in guinea pig caudate membranes. The study focused on two opioid agonists used for treating opioid-dependent patients: methadone and buprenorphine. Selective antagonists were used to generate agonist-selective conditions: TIPP + nor-BNI to measure mu receptors, CTAP + nor-BNI to measure gamma receptors and TIPP + CTAP to measure kappa receptors. The assay was first validated with opioid agonists of known subtype specificity (DAMGO for mu, SNC80 for delta, and U69, 593 for kappa receptors). Methadone-stimulated [(35)S]-GTP-gamma-S binding was mu-specific and less potent and efficacious than etorphine (K(d) = 1,537 nM vs. K(d) = 7.8 nM). Buprenorphine failed to stimulate [(35)S]-GTP-gamma-S binding but inhibited agonist-stimulated [(35)S]-GTP-gamma-S binding. The antagonist-K(i) values (nM) of buprenorphine at mu, delta, and kappa receptors were 0.088 nM, 1.15 nM, and 0.072 nM, respectively. The antagonist-K(i) values (nM) of naloxone at mu, delta, and kappa receptors were 1.39 nM, 25.0 nM, and 11.4 nM, respectively. Autoradiographic studies showed that buprenorphine failed to stimulate [(35)S]-GTP-gamma-S binding in caudate-level rat brain sections but blocked DAMGO-stimulated [(35)S]-GTP-gamma-S binding. In cells expressing the cloned rat mu receptor, buprenorphine was a partial agonist and potent mu antagonist. Administration of buprenorphine to rats produced a long-lasting (>24 h) decrease in mu and kappa2 receptor binding and attenuated mu-stimulated [(35)S]-GTP-gamma-S binding. Viewed collectively, these data indicate that, in this assay system, buprenorphine is a potent mu and gamma receptor antagonist. The clinical implications remain to be elucidated. Synapse 34:83-94, 1999. Published 1999 Wiley-Liss, Inc.