Claudia Gomez

Rapid anxiolytic activity of progesterone and pregnanolone in male rats

The effect of different doses of progesterone (1.0, 3.0, 10.0, 30.0, and 100.0 mg/kg) and pregnanolone (1.0, 3.0, 10.0, and 30.0 mg/kg) upon burying defensive and elevated plus-maze (EPM) tests was investigated in adult male rats and compared with the effects of diazepam (0.25. 0.50, 1.0, and 2.0 mg/kg). All drugs were suspended in a 0.2% methylcellulose solution and administered intraperitoneally 30 min prior to testing. Progesterone and pregnanolone were found to produce anxiolytic-like effects similar to those of diazepam. Thus, at certain doses, both drugs significantly increased the latency for burying and decreased the cumulative burying behavior, without modifying the number of shocks, and increased the time spent in the open arms of the maze, without affecting the spontaneous locomotor activity. These data clearly demonstrate that the defensive burying paradigm is useful to detect the anxiolytic-like properties of pregnanolone. An important finding was that progesterone produces significant behavioral effects 30 min after its administration. This finding suggests a rapid bioconversion of progesterone to its active ring-A reduced metabolites; however, the possibility remains that rapid behavioral effects of progesterone are due to a direct interaction with specific steroid receptors located on the plasma membrane, independently from the gamma-aminobutyric acid(A) (GABA(A)) receptor complex modulation.

Spasmolytic activity of Rosmarinus officinalis L. involves calcium channels in the guinea pig ileum.

ETHNOPHARMACOLOGICAL RELEVANCE Rosmarinus officinalis L. is a plant used around the world for its properties to cure pain in several conditions, such as arthritic and abdominal pain or as an antispasmodic; however, there are no scientific studies demonstrating its spasmolytic activity. Therefore, the aim of the present study was to investigate the effect of an ethanol extract from Rosmarinus officinalis aerial parts and the possible mechanism involved by using rings from the isolated guinea pig ileum (IGPI). MATERIALS AND METHODS The IGPI rings were pre-contracted with potassium chloride (KCl; 60 mM), acetylcholine (ACh; 1 × 10(-9) to 1 × 10(-5)M) or electrical field stimulation (EFS; 0.3 Hz of frequency, 3.0 ms of duration and 14 V intensity) and tested in the presence of the Rosmarinus officinalis ethanol extract (150, 300, 600 and 1 200 μg/mL) or a referenced smooth muscle relaxant (papaverine, 30 μM). In addition, the possible mechanism of action was analyzed in the presence of hexametonium (a ganglionic blocker), indomethacine (an inhibitor of prostaglandins), l-NAME (a selective inhibitor of the nitric oxide synthase) and nifedipine (a calcium channel blocker). RESULTS Rosmarinus officinalis ethanol extract exhibited a significant and concentration-dependent spasmolytic activity on the contractions induced by KCl (CI(50) = 661.06 ± 155.91 μg/mL); ACh (CI(50) = 464.05 ± 16.85 μg/mL) and EFS (CI(50) = 513.72 ± 34.13 μg/mL). Spasmolytic response of Rosmarinus officinalis (600 μg/mL) was reverted in the presence of nifedipine 1 μM, but not in the presence of hexamethonium 0.5mM, indomethacine 1 μM or L-NAME 100 μM. CONCLUSION The present results reinforce the use of Rosmarinus officinalis as antispasmodic in folk medicine. Moreover, it is demonstrated the involvement of calcium channels in this activity, but not the participation of nicotinic receptors, prostaglandins or nitric oxide.

A simplified procedure for the quantitative measurement of neurological deficits after forebrain ischemia in mice.

This study describes a comprehensive method to assess neurological deficits after brain ischemia produced by sequential common carotid artery sectioning (SCAS) in aged mice, and a scale to determine the degree of functional incapacity of ischemic animals. The method involves an initial phase of undisturbed observation and a later manipulative phase during which each animal is subjected to a sequence of very simple manipulations. Sham-operated animals demonstrated 96% survival throughout the study period (72 h), whereas the 24, 48 and 72 h survival rates of SCAS-mice were 48, 38 and 36%, respectively. In the surviving SCAS-mice, we detected a total of 23 neurological alterations throughout the observation period (72 h); the most frequent alterations were: motor incoordination, abnormal body position, hypomobility, decreased body tone and muscular strength, tremor, hunched back, passivity, forelimb flexion and ataxic gait. Based on these alterations, we used a global scale that comprises 10 progressive grades beyond 0 (normal), extending to status 10 (death due to SCAS), with higher scores indicating greater deficit. The median neurological scores for sham-operated animals were 1.36, 1.48 and 1.32 at 24, 48 and 72 h, respectively, whereas total neurological scores in SCAS-mice of 6.1, 6.8 and 7.4, at 24, 48 and 72 h, respectively, were substantially greater than those observed in sham-operated animals. The simplicity of the procedure, herein described, to measure the functional neurological condition of ischemic animals, and the remarkable level of functional impairment produced by SCAS offer the possiblity to test the efficacy of putative stroke therapies and to monitor progress of deficits over time in groups of animals.

Effect of flumazenil and diazepam on transient actions in defensive burying elicited by the social interaction experience in rats.

In the present work, we studied the effects of benzodiazepine (BZ) receptor antagonist, flumazenil, and of the agonist, diazepam, on social interaction-induced transient changes in defensive burying (DB). Enhanced defensive burying was observed after 1.5 min of social interaction experience, while a longer social interaction experience, 15 min, inhibited the expression of burying behavior. Defensive burying and social interaction paradigms have been used for the screening of compounds with anxiolytic potential and, more extensively, to study the neurobiology of anxiety. To elucidate the participation of the BZ receptor on transient changes induced by intervals of social interaction experience, its receptor antagonist, flumazenil (2.5, 5, and 10 mg/kg) was intraperitoneally injected (IP). Flumzenil enhanced in a dose-dependent manner, the blocking effect of the saline IP injection on facilitated DB in 1.5-min social interaction-experienced subjects. In addition, flumazenil enlarged in a dose-dependent manner the blocking effect of saline IP on defensive burying levels in animals exposed to social interaction experience for 15 min. To analyze the presumed participation of the BZ receptor mediating enhanced burying behavior levels in subjects exposed to 1.5 min of social interaction, a suboptimal dose of diazepam (0.25 mg/kg) was administered. Diazepam enhanced the saline IP elicited defensive burying reduction. Results are discussed in terms of the suggested BZ receptor mediation on transient changes in defensive burying elicited by social interaction experience.

Neuroprotective effects of Tilia americana var. mexicana on damage induced by cerebral ischaemia in mice

Abstract Tilia americana var. mexicana (T. americana) is a plant widely used in Mexico for its medicinal properties on the central nervous system. In the present study, we designed a protocol to investigate the neuroprotective effects of non-polar and polar extracts of T. americana on damage induced by cerebral ischaemia in mice. Vehicle or extracts were administered immediately after ischaemia. Functional neurological deficit, survival percentage and infarct area were determined in each experimental group. Results showed that groups treated with non-polar or polar extracts of T. americana had increased survival rate, improved neurological deficits and diminished the infarct area in relation to the ischaemic group. In conclusion, this study confirms the neuroprotective activity of T. americana, suggests a possible synergism between non-polar and polar constituents and supports its potential as a useful aid in the clinical management of stroke.