Assays of homeopathic remedies in rodent behavioural and psychopathological models

The first part of this paper reviews the effects of homeopathic remedies on several models of anxiety-like behaviours developed and described in rodents. The existing literature in this field comprises some fifteen exploratory studies, often published in non-indexed and non-peer-reviewed journals. Only a few results have been confirmed by multiple laboratories, and concern Ignatia, Gelsemium, Chamomilla (in homeopathic dilutions/potencies). Nevertheless, there are some interesting results pointing to the possible efficacy of other remedies, and confirming a statistically significant effect of high dilutions of neurotrophic molecules and antibodies. In the second part of this paper we report some recent results obtained in our laboratory, testing Aconitum, Nux vomica, Belladonna, Argentum nitricum, Tabacum (all 5CH potency) and Gelsemium (5, 7, 9 and 30CH potencies) on mice using ethological models of behaviour. The test was performed using coded drugs and controls in double blind (operations and calculations). After an initial screening that showed all the tested remedies (except for Belladonna) to have some effects on the behavioural parameters (light–dark test and open-field test), but with high experimental variability, we focused our study on Gelsemium, and carried out two complete series of experiments. The results showed that Gelsemium had several effects on the exploratory behaviour of mice, which in some models were highly statistically significant (p < 0.001), in all the dilutions/dynamizations used, but with complex differences according to the experimental conditions and test performed. Finally, some methodological issues of animal research in this field of homeopathy are discussed. The “Gelsemium model” – encompassing experimental studies in vitro and in vivo from different laboratories and with different methods, including significant effects of its major active principle gelsemine – may play a pivotal rule for investigations on other homeopathic remedies.

Keywords

Behaviour - Anxiety - Psychopatology - Nervous system - Catalepsy - Rodent models - Thigmotaxis - Animal studies - Ignatia - Chamomilla - Gelsemium - Ultra-high dilutions - Review - Homeopathic remedies

• References

• 1 Young R., Johnson D.N. A fully automated light/dark apparatus useful for comparing anxiolytic agents. Pharmacol Biochem Behav 1991; 40: 739-743.
• 2 Bourin M., Petit-Demouliere B., Dhonnchadha B.N., Hascoet M. Animal models of anxiety in mice. Fundam Clin Pharmacol 2007; 21: 567-574.
• 3 Griebel G., Belzung C., Misslin R., Vogel E. The free exploratory paradigm: an effective method for measuring neophobic behaviour in mice and testing potential neophobia reducing drugs. Behav Pharmacol 1993; 4: 637-644.
• 4 Clement Y., Joubert C., Kopp C. et al. Anxiety in mice: a principal component analysis study. Neural Plast 2007 10.1155/2007/35457
• 5 Belzung C., Griebel G. Measuring normal and pathological anxiety-like behaviour in mice: a review. Behav Brain Res 2001; 125: 141-149.
• 6 Hascoet M., Bourin M., Dhonnchadha B.A. The mouse light–dark paradigm: a review. Prog Neuropsychopharmacol Biol Psychiatry 2001; 25: 141-166.
• 7 Bourin M., Hascoet M. The mouse light/dark box test. Eur J Pharmacol 2003; 463: 55-65.
• 8 Chen S.W., Min L., Li W.J., Kong W.X., Li J.F., Zhang Y.J. The effects of angelica essential oil in three murine tests of anxiety. Pharmacol Biochem Behav 2004; 79: 377-382.
• 9 Simon P., Dupuis R., Costentin J. Thigmotaxis as an index of anxiety in mice. Influence of dopaminergic transmissions. Behav Brain Res 1994; 61: 59-64.
• 10 Tetau J., Tetau M. Pharmacologie et psycho-pharmacologie de Thuya. Ann Homéop Fr 1960; 2: 669-678.
• 11 Julian O.A., Launay J. Psycho-pathological test on animals by reserpine and Cicuta virosa, according to the homeopathic laws of analogy and identity. J Am Inst Homeopath 1966; 59: 155-164.
• 12 Binsard A.M. Étude psycho-pharmacologique de diluitions homéopathiques d'Ignatia. Ann Homéop Fr 1978; 20: 313-321.
• 13 Binsard A.M. Étude psycho-pharmacologique d'Ignatia et rapprochement avec un autre polychreste. Ann Homéop Fr 1979; 21: 369-378.
• 14 Binsard A.M., Guillemain J., Platel A., Savini E.C., Tetau M. Étude psycho-pharmacologique de dilutions homéopathiques de Gelsemium et d'Ignatia. Ann Homéop Fr 1980; 22: 35-50.
• 15 Guillemain J., Huguet F., Binsard A.M., Tetau M., Narcisse G. Action anti-convulsivante expérimentale de diluitions d'Ignatia chez la souris. Ann Homéop Fr 1981; 23: 35-41.
• 16 Sukul N.C., Bala S.K., Bhattacharyya B. Prolonged cataleptogenic effects of potentized homoeopathic drugs. Psychopharmacology (Berl) 1986; 89: 338-339.
• 17 Sukul N.C., Sukul A. High dilution effects: physical and biochemical basis. Dordrecht: Kluwer; 2003.
• 18 Guillemain J., Rousseau A., Dorfman P., Tetau M. Recherche en psychopharmacologie. Cahiers de Biotherapie 1989; 103: 53-66.
• 19 Sukul N.C., Batuev A.S., Sabanov V., Kourzina N.P. Neuronal activity in the lateral hypothalamus of the cat the medial frontal cortex of the rat in response to homoeopathic drugs. Indian Biol 1991; 23 (02) 17-21.
• 20 Cristea A., Teodorescu-Negres S., Darie V. Chamomilla homeopathic dilution effect on central nervous system. An experimental pharmacological study. In: Bastide M. Signals and Images. 1997. Dordrecht (NL): Kluwer Acad. Publ.; 171-178
• 21 Sukul A., Sinhabau S.P., Sukul N.C. Reduction of alcohol induced sleep time in albino mice by potentized Nux vomica prepared with 90% ethanol. Br Hom J (Homeopathy) 1999; 88: 58-61.
• 22 Sukul N.C., Ghosh S., Sinhababu S.P., Sukul A. Strychnos nux-vomica extract and its ultra-high dilution reduce voluntary ethanol intake in rats. J Altern Complement Med 2001; 7: 187-193.
• 23 Bousta D., Soulimani R., Jarmouni I. et al. Neurotropic, immunological and gastric effects of low doses of Atropa belladonna L., Gelsemium sempervirens L. and Poumon histamine in stressed mice. J Ethnopharmacol 2001; 74: 205-215.
• 24 Ruiz-Vega G., Perez-Ordaz L., Leon-Hueramo O., Cruz-Vazquez E., Sanchez-Diaz N. Comparative effect of Coffea cruda potencies on rats. Homeopathy 2002; 91: 80-84.
• 25 de Paula C.C., D'Almeida V., Pedrazzolli-Neto M. et al. Therapeutic and pathogenetic animal models for Dolichos pruriens . Homeopathy 2006; 95: 136-143.
• 26 Epstein O.I., Pavlov I.F., Shtark M.B. Improvement of memory by means of ultra-low doses of antibodies to S-100B antigen. Evid Based Complement Alternat Med 2006; 3: 541-545.
• 27 Pankova T.M., Starostina M.V., Shtark M.B., Epstein O.I. Neuroprotective effect of ultra-low doses of antibodies against S100 protein in neuroblastoma culture during oxygen and glucose deprivation. Bull Exp Biol Med 2007; 144: 288-290.
• 28 da Silva Rocha M.P., Soares F.M., Martini L.C., Bonamin L.V. Behaviour of rats treated with Rhus toxicodendron 200CH. Int J High Dil Res 2008; 7 (22) 3-6.
• 29 Pinto S.A., Bohland E., Coelho C.P., Morgulis M.S., Bonamin L.V. An animal model for the study of Chamomilla in stress and depression: pilot study. Homeopathy 2008; 97: 141-144.
• 30 Magnani P., Conforti A., Bellavite P. Effects of homeopathic drugs on the anxiety-like behaviour in mice. In: Van Wassenhoven M. Proceedings of 63rd World Congress of the Liga Medicorum Homeopathica Internationalis. 2008. Ostend: LMHI Edition; (CD-ROM)
• 31 Bellavite P., Magnani P., Zanolin M.E., Conforti A. Homeopathic doses of Gelsemium sempervirens improve the behavior of mice in response to novel environments. Evid Based Complement Alternat Med 2009 10.1093/ecam/nep139
• 32 Ramos A., Pereira E., Martins G.C., Wehrmeister T.D., Izidio G.S. Integrating the open field, elevated plus maze and light/dark box to assess different types of emotional behaviors in one single trial. Behav Brain Res 2008; 193: 277-288.
• 33 Guillemain J., Huguet F., Segouin C., Bakri Logeais F., Tetau M., Narcisse G. Liaisons in-vitro de diluitions d'Ignatia et Strychninum aux récepteurs glycinergiques: recherche de spécificité. Ann Homéop Fr 1981; 23: 25-33.
• 34 Sukul N.C. Anticataleptic effect of Agaricus muscaris at ultra high dilutions. Indian J Physiol Allied Sci 1995; 49: 52-58.
• 35 Pinheiro S.H., Zangrossi Jr. H., Del-Ben C.M., Graeff F.G. Elevated mazes as animal models of anxiety: effects of serotonergic agents. An Acad Bras Cienc 2007; 79: 71-85.
• 36 Ruiz-Vega G., Perez-Ordaz L., Proa-Flores P., Aguilar-Diaz Y. An evaluation of Coffea cruda effect on rats. Brit Hom J (Homeopathy) 2000; 89: 122-126.
• 37 Ruiz-Vega G., Perez-Ordaz L., Cortes-Galvan L., Juarez G. A kinetic approach to caffeine—Coffea cruda interaction. Homeopathy 2003; 92: 19-29.
• 38 Ruiz-Vega G., Poitevin B., Perez-Ordaz L. Histamine at high dilution reduces spectral density in delta band in sleeping rats. Homeopathy 2005; 94: 86-91.
• 39 Ruiz-Vega G., Estevez-Delgado G. Non-linearity modeling of ultra-dilutions: the histamine disturbances case. In: Bonamin L.V. Signals and Images. Contributions and Contradictions about High Dilution Research. 2008. Dordrecht: Springer Science; 67-82
• 40 Linde K., Jonas W.B., Melchart D., Worku F., Wagner H., Eitel F. Critical review and meta-analysis of serial agitated dilutions in experimental toxicology. Hum Exp Toxicol 1994; 13: 481-492.
• 41 Witt C.M., Bluth M., Albrecht H., Weisshuhn T.E., Baumgartner S., Willich S.N. The in vitro evidence for an effect of high homeopathic potencies—a systematic review of the literature. Complement Ther Med 2007; 15: 128-138.
• 42 Van Wijk R., Albrecht H. Classification of systems and methods used in biological basic research on homeopathy. Homeopathy 2007; 96: 247-251.
• 43 Van Wijk R. The in vitro evidence for an effect of high homeopathic potencies—a systematic review of the literature. Complement Ther Med 2007; 15: 139-141.
• 44 Venard C., Boujedaini N., Belon P., Mensah-Nyagan A.G., Patte-Mensah C. Pharmacological modulators of the glycinergis system regulate allopregnanolone biosynthesis in the rat spinal cord. In: Panzica G.C., Gotti S. Proceedings of the 4th International Meeting on Steroids and Nervous System. 2007. Madrid: Trabajos del instituto cajal; 262
• 45 Venard C., Boujedaini N., Belon P., Mensah-Nyagan A.G., Patte-Mensah C. Regulation of neurosteroid allopregnanolone biosynthesis in the rat spinal cord by glycine and the alkaloidal analogs strychnine and gelsemine. Neuroscience 2008; 153: 154-161.
• 46 Peredery O., Persinger M.A. Herbal treatment following post-seizure induction in rat by lithium pilocarpine: Scutellaria lateriflora (Skullcap), Gelsemium sempervirens (Gelsemium) and Datura stramonium (Jimson Weed) may prevent development of spontaneous seizures. Phytother Res 2004; 18: 700-705.
• 47 Treit D., Fundytus M. Thigmotaxis as a test for anxiolytic activity in rats. Pharmacol Biochem Behav 1988; 31: 959-962.
• 48 Lamprea M.R., Cardenas F.P., Setem J., Morato S. Thigmotactic responses in an open-field. Braz J Med Biol Res 2008; 41: 135-140.
• 49 Leppanen P.K., Ravaja N., Ewalds-Kvist S.B. Twenty-three generations of mice bidirectionally selected for open-field thigmotaxis: selection response and repeated exposure to the open field. Behav Processes 2006; 72: 23-31.
• 50 Hodgson S., Hofford R., Buckman S., Wellman P., Eitan S. Morphine-induced stereotyped thigmotaxis could appear as enhanced fear and anxiety in some behavioural tests. J Psychopharmacol 2009
• 51 Kallai J., Makany T., Csatho A. et al. Cognitive and affective aspects of thigmotaxis strategy in humans. Behav Neurosci 2007; 121: 21-30.
• 52 Crawley J.N. Exploratory behavior models of anxiety in mice. Neurosci Biobehav Rev 1985; 9: 37-44.
• 53 Choleris E., Thomas A.W., Kavaliers M., Prato F.S. A detailed ethological analysis of the mouse open field test: effects of diazepam, chlordiazepoxide and an extremely low frequency pulsed magnetic field. Neurosci Biobehav Rev 2001; 25: 235-260.
• 54 Mitchell E.A., Herd M.B., Gunn B.G., Lambert J.J., Belelli D. Neurosteroid modulation of GABAA receptors: molecular determinants and significance in health and disease. Neurochem Int 2008; 52: 588-595.
• 55 Belelli D., Herd M.B., Mitchell E.A. et al. Neuroactive steroids and inhibitory neurotransmission: mechanisms of action and physiological relevance. Neuroscience 2006; 138: 821-829.
• 56 Ahboucha S., Coyne L., Hirakawa R., Butterworth R.F., Halliwell R.F. An interaction between benzodiazepines and neuroactive steroids at GABA A receptors in cultured hippocampal neurons. Neurochem Int 2006; 48: 703-707.
• 57 Pinna G., Costa E., Guidotti A. Fluoxetine and norfluoxetine stereospecifically and selectively increase brain neurosteroid content at doses that are inactive on 5-HT reuptake. Psychopharmacology (Berl) 2006; 186: 362-372.
• 58 Izumi Y., Murayama K., Tokuda K., Krishnan K., Covey D.F., Zorumski C.F. GABAergic neurosteroids mediate the effects of ethanol on long-term potentiation in rat hippocampal slices. Eur J Neurosci 2007; 26: 1881-1888.
• 59 Venard C., Boujedaini N., Mensah-Nyagan A.G., Patte-Mensah C. Comparative analysis of Gelsemine and Gelsemium sempervirens activity on neurosteroid allopregnanolone formation in the spinal cord and limbic system. Evid Based Complement Alternat Med 2009 10.1093/ecam/nep083
• 60 Patte-Mensah C., Kibaly C., Boudard D. et al. Neurogenic pain and steroid synthesis in the spinal cord. J Mol Neurosci 2006; 28: 17-31.
• 61 Strous R.D., Maayan R., Weizman A. The relevance of neurosteroids to clinical psychiatry: from the laboratory to the bedside. Eur Neuropsychopharmacol 2006; 16: 155-169.
• 62 Girdler S.S., Klatzkin R. Neurosteroids in the context of stress: implications for depressive disorders. Pharmacol Ther 2007; 116: 125-139.
• 63 Ahboucha S., Butterworth R.F. The neurosteroid system: an emerging therapeutic target for hepatic encephalopathy. Metab Brain Dis 2007; 22: 291-308.
• 64 Griffin L.D., Gong W., Verot L., Mellon S.H. Niemann-Pick type C disease involves disrupted neurosteroidogenesis and responds to allopregnanolone. Nat Med 2004; 10: 704-711.
• 65 Mellon S.H., Gong W., Schonemann M.D. Endogenous and synthetic neurosteroids in treatment of Niemann-Pick type C disease. Brain Res Rev 2008; 57: 410-420.
• 66 Marx C.E., Trost W.T., Shampine L.J. et al. The neurosteroid allopregnanolone is reduced in prefrontal cortex in Alzheimer's disease. Biol Psychiatry 2006; 60: 1287-1294.
• 67 Brown V., Ennis M. Flow-cytometric analysis of basophil activation: inhibition by histamine at conventional and homeopathic concentrations. Inflamm Res 2001; 50 (02) S47-S48.
• 68 Belon P., Cumps J., Ennis M. et al. Histamine dilutions modulate basophil activation. Inflamm Res 2004; 53: 181-188.
• 69 Eizayaga F.X., Aguejouf O., Belon P., Doutremepuich C. Platelet aggregation in portal hypertension and its modification by ultra-low doses of aspirin. Pathophysiol Haemost Thromb 2005; 34: 29-34.
• 70 Bellavite P., Conforti A., Pontarollo F., Ortolani R. Immunology and homeopathy. 2. Cells of the immune system and inflammation. Evid Based Complement Alternat Med 2006; 3: 13-24.
• 71 Chirumbolo S., Brizzi M., Ortolani R., Vella A., Bellavite P. Inhibition of CD203c membrane up-regulation in human basophils by high dilutions of histamine: a controlled replication study. Inflamm Res 2009 10.1007/s00011-009-0044-4
• 72 Roy R., Tiller W., Bell I.R., Hoover M.R. The structure of liquid water. Novel insights from materials research; potential relevance to homeopathy. Mater Res Innovat 2005; 9: 98-103.
• 73 Mastrangelo D. Hormesis, epitaxy, the structure of liquid water, and the science of homeopathy. Med Sci Monit 2007; 13: SR1-SR8.
• 74 Chaplin M.F. The memory of water: an overview. Homeopathy 2007; 96: 143-150.
• 75 Anick D.J., Ives J.A. The silica hypothesis for homeopathy: physical chemistry. Homeopathy 2007; 96: 189-195.
• 76 Weingartner O. The nature of the active ingredient in ultramolecular dilutions. Homeopathy 2007; 96: 220-226.
• 77 Ball P. Water – an enduring mystery. Nature 2008; 452: 291-292.
• 78 Demangeat J.L. NMR water proton relaxation in unheated and heated ultrahigh aqueous dilutions of histamine: evidence for an air-dependent supramolecular organization of water. J Mol Liq 2008 10.1016/j.molliq.2008.07.013
• 79 Poitevin B. The continuing mystery of the memory of water. Homeopathy 2008; 97: 39-41.
• 80 Bellavite P., Signorini A. The emerging science of homeopathy. Berkeley (CA): North Atlantic; 2002.
• 81 Bellavite P. Complexity science and homeopathy. A synthetic overview. Homeopathy 2003; 92: 203-212.
• 82 Bellavite P., Ortolani R., Pontarollo F., Pitari G., Conforti A. Immunology and homeopathy. 5. The rationale of the ‘Simile’. Evid Based Complement Alternat Med 2007; 4: 149-163.