Mice behavioural models with pooled data analysis of Gelsemium studies and new findings about Ignatia and Aconitum

Authors

  • Paolo Bellavite University of Verona (I)
  • Paolo Magnani University of Verona (I)
  • Marta Marzotto University of Verona (I)
  • Mirko Cristofoletti University of Verona (I)
  • Mariaelisabetta Zanolin University of Verona (I)
  • Anita Conforti University of Verona (I)

Abstract

Objective: We investigated the effects of three different homeopathic medicines in several dilutions/dynamizations on mice, using validated models which explore anxiety-like and emotional symptoms. Two complete series of investigations were performed in order to assess the activity of Gelsemium sempervirens; furthermore, we investigated Ignatia amara and Aconitum napellus in the same model systems. Methods: Mice of CD1 strain were randomized in different cages (minimum 8 mice per treated group in each experiment) and treatment solutions were coded in such a way that all protocols were carried out fully in blind. The indicated compounds at various centesimal dilutions/dynamizations, a control solution (the solvent vehicle of drugs, which was succussed before administration) or the reference drugs diazepam (1 mg/kg body weight) or buspirone (5 mg/kg body weight) diluted in the same succussed solvent were delivered intraperitoneally (0.3 ml/mice) for 9 days. A series of changes of animal behavior were assessed by the Light-Dark (LD) choice test and the Open-Field (OF) exploration test. Two series of studies with little technical differences, exploiting a total of 14 separate experiments, were carried out with Gelsemium, five complete experiments with Ignatia and four complete experiments with Aconitum. Results: In both series of experiments Gelsemium showed anxiolytic-like effects using both OF test (permanence and movement in centre area of field) and LD test (time spent in lit area and number of light-dark transitions). However, due to high variability of animal responses and possibly to some minor differences in protocols, those effects reached the threshold of statistical significance only in OF in the first series and only in LD in the second series. Cumulative analysis of the two series demonstrated a highly significant (p<0.0001) effect of Gelsemium 5CH,7CH, and 30CH in OF parameters and of Gelsemium 5CH, 9CH, and 30CH in LD parameters. In OF the effect of Gelsemium was evident at variance with the effect of the standard drugs diazepam and buspirone, suggesting an increase in exploratory behavior and a decrease in thigmotaxis or in neophobia, instead of a pure anxiolytic effect; in LD test the effects of Gelsemium were in the same direction as those of benzodiazepines. Ignatia showed statistically significant effects only in the LD test parameters (peak at 9CH dilution/dynamization), while Aconitum anxiolytic-like effects were evident only in one experiment where the basal anxiety level of mice was very high. In addition, we observed a tendency of Ignatia to decrease the number of urinations and of Aconitum to decrease the number of stools produced by animals, an effect that was in the same direction as the activity of diazepam. Homeopathic medicines did not alter the general locomotion of mice in the OF, indicating that their effect was actually anxiolytic-like and not sedative, while buspirone significantly inhibited this parameter. Conclusions: Pharmacological effects of highly diluted/dynamized medicines on behavioural and emotional symptoms are clearly detectable also in experimental animals. Among the three tested remedies, in these model paradigms Gelsemium showed the highest activity, which was statistically significant as compared with pure solvent even in solutions diluted beyond the Avogadro constant. The effects of different medicines on the anxiety-like symptoms were qualitatively different and differed also from those of benzodiazepines and of serotonin partial agonists. These experiences raise some technical issues related to the animal models and to the possible translation to homeopathy in humans.

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