Biological Action of Homeopathy Medicine

Mechanism of biological action(s) of the potentized homeopathic remedies can be scientifically explained through a working hypothesis

One of the main reasons for the lack of acceptance of homeopathy as a scientific mode of treatment is the absence of a clear understanding of its mechanism of action, particularly of the ultra-high diluted potentized remedies. At the present state of our knowledge, the mechanism of action of the potentized homeopathic remedies diluted beyond Avogadro's limit is extremely difficult to understand and explain. But here is a working hypothesis that can throw light on the possible mechanism of biological action of the potentized homeopathic remedies with concrete supportive evidences. In the beginning, let me focus on the intricacies that come in the way of a scientific explanation of the mechanism of action of the homeopathic remedies, particularly those diluted above the Avogadro's limit. Firstly, to understand the mechanism of action of the potentized homeopathic drugs, one has to satisfactorily answer the problem of how the medicinal property of the original drug substance can be transferred to and retained by the ethanolic vehicle. Several working hypotheses have been proposed in this aspect (Anaganostatos et al., 1998; Allegre et al., 1989; Bellavite and Signorini, 2002; Elia and Niccoli 1999; Lo, 1996; Anick and Ives, 2007; Rey et al., 2007). The leading current proposal for the mode of action of such "ultra molecular" dilutions is that water (and aquatic ethanol as well) is capable of storing information relating to substances with which it has been in contact and subsequently can transmit this information to pre-sensitized biological system. The process is believed to be mediated by structural modification of water (or the aquatic ethanol, the "vehicle" of the homeopathic remedies), analogous to storage of information by magnetic media. Such information is retained in physical, rather than chemical form (Bellavite and Signorini, 2002). Several aspects of the "water structure" and "memory of water" have been critically analyzed recently by Chaplin (2007). Results of studies conducted on nature of molecular clustering in water solutions showed that as a solution is made more and more dilute, larger and very stable aggregates develop (Samal and Geckeler, 2001). This would mean that residual molecular clusters of the original substance may be present in homeopathic dilutions. Interestingly, alcohol also forms clusters with water (Wisnieski et al., 2001), which would further support the homeopathic analogy. Being an associative liquid, that is having hydrogen bonds between molecules, it is thus possible that alcohol can form its very own clusters. In fact, the presence of alcohol in water may actually be favorable for the moderation of succussion energy (by increasing vapor pressure). Andersson et al (1997) and Markovic et al (1999) were able to create individual clusters, using sudden evaporative cooling. According to these authors, cluster mixtures can generate many different isomeric forms in different geometric forms, and each isomer can represent a unique bit of information. Thus it is possible that millions of different "information bits" can be produced which may be "biologically active". Hameroff and Penrose (2001) have proposed that microtubules in our brain are the seat of our conscious mind and consist of quantum micro switches (protein qubits), which contain "pure" ordered water. The microtubules interconnect every cell in a multicellular organism possibly being the means to extend the brain's consciousness throughout the entire body. These authors further suggest that this microtubule network can explain the mind-body interactions, such as psycho-somatic pain. There are many research articles that suggest the role of cell cytoskeleton in genome regulation in cancer. Further there are evidences that support the fact that internal microtubules of the cell control mitosis (cell division), regulate the genes, decide which gene to turn on and so forth not only in terms of differentiation in development but also in health and in the steady-state. They believe that consciousness, the microtubules and quantum coherence play an essential role in health and diseases. Thus, in fine, the water cluster model and microtubule network model are comparable with Hahnemann's concept of vital force in which a biological network offers all the action/reaction properties. Additionally, Hameroff's (2001) work suggests that the quantum state of the microtubule's elemental building block can be altered by an 'ordered' water structure attached to the external side of the microtubules. The water cluster(s), which is (are) specific to the problems, can simply throw the right switch or switches to correct the error in the network. Thus molecular architecture of water has a key role to play in understanding homeopathic mechanism of action and has therefore been vigorously studied (Dantas et al., 2007; Elia et al., 2007; Anick and Ives (2007).

The next part of the mechanism that should be addressed is how the tiny drop of a potentized remedy or a few sugar globules soaked with the remedy can trigger the recovery process of the disease/abnormal symptoms. There are several hypotheses based on in vivo and in vitro studies with various animal models in relation to various immunological, cardiovascular and molecular aspects and various kinds of hypotheses like "hormesis", "allergic reaction", or "immunological responses" etc. have been proposed (see The HomBRex database, available at: http://www.carstens-stiftung.de/hombrex ). These hypotheses have their apparent merits, but also suffer from some inadequacies in explaining the action of homeopathic drugs in unicellular organisms like protozoans or bacteria. Any acceptable hypothesis has to be able to explain the mechanism of action universally in all kinds of organisms where homeopathic remedies have been reported to act. Khuda-Bukhsh (1997, 2003, 2006, 2009) proposed a hypothesis based on various scientific evidences that potentized homeopathic drugs act through regulation of relevant gene expression. The possible pathways and sites of action have also been discussed by this group (Khuda-Bukhsh, 1997; Mallick et al., 2003; Pathak et al., 2006; 2007; Khuda-Bukhsh and Pathak, 2008). According to this hypothesis, homeopathic remedies carry specific "signals"/"information" that can be identified by specific receptors of the cells. These "signals" can act as a "trigger" for turning "on" or "off" some relevant genes, initiating a cascade of gene actions to alter and correct the gene expressions that went wrong to produce the disorder/disease. Presumably, the homeopathic drugs are mediated through cytokine responses. Thus, administration of a potentized homeopathic drug can elicit response in suitable signal proteins and can either up-regulate or down-regulate such signal proteins to bring back the recovery of the patient to normal health (Khuda-Bukhsh et al., 2009). For the same reason a global microarray after a homeopathic drug administration in a disease/abnormal state should provide evidence for modulated expression of a large number of genes that has actually been reported recently (D Oliveira et al., 2008).

In higher eukaryotes like mammals, the regulation of gene expression is, however, a very complex phenomenon. In principle, the expression of genes might be regulated at any one of several stages. At least five control points can be distinguished that form the series, through which gene expression is regulated (see Lewin 2004): Activation of structure - Initiation of transcription - Processing the transcript - Transport to cytoplasm - Translation of mRNA. Many eukaryotic genes have multiple regulatory binding sites and are controlled by more number of regulatory proteins (see Watson et al., 2004). Enhancers bind regulators responsible for activating the gene at a given time and place. Alternative enhancers bind different groups of regulators and control expression of the same gene at different times and places in response to different signals. Some regulators bind sites far from the genes they control, by repressor proteins. Repressors work by a variety of ways, including the way which is called "gene silencing". Modification to regions of chromatin keep genes in sometimes large stretches of DNA "switched off". Similarly, there are "activators", which do not always have well-defined structure. Activators recruit the transcriptional machinery to the gene, and sometimes recruit nucleosome modifiers that help the transcription machinery bind at the promoter. Activators are known to work synergistically to integrate signals. Synergy is critical for signal integration by activators. Each signal is communicated to the gene by a separate activator (signal recognition particle). Signals are often communicated to transcriptional regulators through signal transduction pathways. There are various ways that signals are detected by a cell and communicated to a gene. It can also refer to the "information" as it passes from detection of the ligand to the regulators that directly control the genes- that is, as it passes along a signal transduction pathway. In a signal transduction pathway, the initiating ligand is typically detected by a specific cell surface receptor: the ligand binds to an extracellular domain of the receptor and this binding is communicated to the intracellular domain. From there the signal is relayed to the relevant regulator, often through a cascade of kinases. Generally binding of ligand alters the shape (and thus activity) of the intracellular domain. Alternatively the ligand can act simply to bring together two or more receptor chains, allowing interaction domains of those receptors to activate each other (see Watson et al., 2004). However, how a homeopathic drug can elicit response in a cell receptor and can bind with the receptor is not precisely known. These are the areas that should be more specifically searched. Our research points out a possibility that homeopathic drug can have interaction with some nano-particles that can tag onto some proteins. That can make it possible for some drug-associated nano-particles to get attached to some proteins, which then act as the ligand (Bhattacharyya et al., 2008). However, this is only speculative at this stage of our knowledge and needs validation by further research. A recent finding that nano-particles of metals can be traced in ultra-high diluted homeopathic remedies through high powered electron microscopes vindicates our hypothesis.

Likewise, there are STAT and MAP kinase pathways activated by ligand (a cytokine) in which phosphorylation and dephosphorylation of different kinases occur to carry forward the information to produce activities in genes downstream as a cascade of reactions. Perhaps signals of potentized homeopathic drugs are carried through this method (Khuda-Bukhsh, 2009; Khuda-Bukhsh et al., 2009). Gene expression can also be controlled by "signals" received by a cell from its environment. For example, the presence of sugar lactose activates the transcription of the lac operon in E. coli, while viral infection activates the expression of β-interferon gene in mammals (see Watson et al., 2004). Interestingly, in our recent experiment on E. coli subjected to low dose of arsenic, Arsenicum Album 30C showed modulatory effect in their glucose uptake (results unpublished). Generally speaking, the strategies that are used to instruct genetically-identical cells to express distinct sets of genes and thereby perform different sets of function include m-RNA localization, cell-to-cell contact and signaling through the diffusion of a secreted signaling molecule. Microarray assays can thus help further in the analysis of gene expression profiles of experimental organisms administered micro doses of the homeopathic drug.

In fine, the reasons and circumstantial evidences that support gene regulatory hypothesis are:

• The use of various state-of-the art techniques has now quite convincingly proved the efficacy of several potentized homeopathic drugs diluted beyond Avogadro's limit in modulating significantly many parameters of study. These parameters are considered scientifically acceptable and dependable. The potentized homeopathic remedies are capable of modulating such parameters, while the succussed alcohol (placebo) can not. Further, the homeopathic drug can bring about changes in multiple parameters at any given time frame, which without a cascade of gene actions is not possible.
• The homeopathic remedies are diluted to such an extent that there can not be physical existence of a single molecule of the original drug substance. Therefore it is simply not possible for the remedy to react chemically with all the cells undergoing some degree of quantifiable/visible changes at the patho-physiological level. But there are many evidences that demonstrate modulations by the potentized homeopathic remedy (diluted beyond Avogadro's limit) in regard to almost every endpoint studied.
• This leaves us with the only possibility that the homeopathic drug carries molecular imprints or definite signal/information of the original drug molecule that can be deciphered by the cells' receptors. If one knows a particular language, no matter if the size of the script is small or large, the message will carry the same meaning. Therefore, if they indeed carry some signals, and the cells can perceive that, then there should be a reactive process going on in the transduction of the signal to the target organ. Most of the parameters of study, particularly the expression of signal proteins, are indicators of the fact that their genes have been expressed in terms of either up-regulation or down regulation, as the case may be, that is good evidence of their ability to exert influence on the regulatory genes of these signal proteins; since the expression of these signal proteins and some other specific receptor proteins (like AhR or PCNA) have also been tested positive by some other confirmatory tests like immunohistochemical localization as well. Further, the ability of the homeopathic drugs in bringing about structural alterations at the sub-cellular levels in some vital organs (like liver) has been evidenced by electron microscopic studies as well, that would also speak for its ability to tackle matters at the molecular level.
• The use of several modern techniques involving monoclonal antibodies for the detection of these signal proteins through immunoblot experiments are currently accepted to be quite dependable scientific tool for ascertaining gene modulating effects of any drug.
• Nanoparticles have been shown to have effect on the physico-chemical property of the homeopathic dilutions (Bhattacharyya et al., 2008), that may indicate some role of the nano-particles during the potentization process of the homeopathic drugs.
• All parameters of study tested so far are strictly gene controlled.
• Homeopathic dilutions have been noted to have demonstrable and reproducible action on plants, and bacteria, which lack any central nervous system.
• And finally, the simultaneous treatment of Actinomycin D, a transcription blocker, the action of a potentized homeopathic remedy fails (Dutta et al., 1999; Chakrabarti et al., 2001).

Thus, our extensive research on homeopathy yielded much suggestive information for understanding the biological part of the mechanism of action. More in-depth research, particularly by other independent researchers, to test and verify our research findings will be highly solicited, because validation of results obtained by us can indeed open up newer avenues, paving the way for a better understanding of the mechanism and pathways of biological action of the potentized homeopathic remedies in near future, and to establish the "gene regulation" hypothesis as universally acceptable one.

References

1. Allegre CJ, Provost A, Jaupart C. Oscillatory zoning: a pathological case of crystal growth. Nature 1989; 294: 223-228.
2. Anagnostatos GS, Pissis P, Viras K, et al. Theory and experiments on high dilutions. In: Homoeopathy - a critical appraisal (Eds: Ernst E and Hahn EG), Butterworth-Heinemann, Reed Educational and Professional Publishers Ltd., 1998; 153-166.
3. Andersson PU, Tomsic A, Andersson MB,et al. Emission of small fragments during water cluster collisions with graphite surface. Chem Phys Lett 1997; 279: 100-106.
4. Anick DJ, Ives JA. The silica hypothesis for homeopathy:physical chemistry. Homeopathy 2007; 96: 189-195.
5. Bellavite P, Signorini A. The Emerging Science of Homeopathy . North Atlantic, Berkeley, 2002.
6. Bhattacharyya SS, Mandal SK, Biswas R et al. In vitro studies demonstrate anticancer activity of an alkaloid of plant Gelsemium semipervirens. Exp Biol Med 2008; 233: 1591-1601.
7. Chakrabarti, J., Biswas SJ, Khuda-Bukhsh AR. Cytogenetical effects of sonication in mice and their modulations by actinomycin D and a homeopathic dug., Arnica 30. Indian J Exp Biol 2001: 39: 1235-1242.
8. Chaplin MF. The memory of water: an overview. Homeopathy 2007; 96: 143-150.
9. Doliviera de Oliveira CC, de Oliveira SM, Goes VM, Probst CM, Krieger MA, Buchi Dde F. Gene expression profiling of macrophages following mice treatment with an immunomodulator medication.. J Cell Biochem . 2008; 104: 1364-1377.
10. Dantas F, Fisher P, Walach H, et al. A systematic review of the quality of homeopathic pathogenetic trials published from 1945 to 1995.Homeopathy 2007; 96: 4-16.
11. Dutta S., Malick P., Khuda-Bukhsh AR. Of a potentized homepahic drug (Arsenicum Album 30) in reducing genotoxic effects produced by arsenic trioxide in mice. .II. Comparative efficacy of an antibiotic, actinomycin D alone and in combination with either of two microdoses. Comp Ther Med 1999, 7: 156-163.
12. Elia V, Napoli E, Germano R. The "Memory of Water": an almost deciphered enigma. Dissipative structures in extremely dilute aqueous solutions. Homeopathy 2007; 96:163-169.
13. Elia V, Niccoli M. Thermodynamics of extremely diluted aqueous solutions. Ann N Y Acad Sci. 1999; 879: 241-248.
14. Hameroff S. Departments of Anesthesiology and Psychology, Associate Director, Center for Consciousness Studies, The University of Arizona, Tucson, Arizona, 2001. Available at http://www.consciousness.arizona.edu/hameroff
15. Khuda-Bukhsh AR. Some homeopathic drugs as radio-protective agents in X-irradiated mice. Perspectives in Cytology and Genetics (eds. G.K. Manna and U Sinha), 1986, 5: 407-412.
16. Khuda-Bukhsh AR : Potentized homeopathic drugs act through regulation of gene expression : a hypothesis to explain their mechanism and pathways of action in vivo. Comp Ther Med 1997; 5: 43-46.
17. Khuda-Bukhsh AR : Towards understanding molecular mechanisms of action of homeopathic drugs:An over view. Mol Cell Biochem 2003; 253: 339-345.
18. Khuda-Bukhsh AR, Laboratory research in homeopathy:pro. Integr Cancer Ther 2 Khuda-Bukhsh AR. Mice as a model for homeopathy research. Homeopathy. doi: 10.1016/j.homp.2009.09.007 available online at http://www.sciencedirect.com ; 006; 5: 1-14.
19. Khuda-Bukhsh AR, Bhattacharryya SS, Paul S, Datta S, Boujedaini N, Belon P. Modulation of signal proteins: a plausible mechanism to explain how a potentized drug Secale cor 30C diluted beyoned Avogadro's limit combat skin papilloma in mice Evid BasedComplememt Alternet Med 2009, 1-12, doi:10.1093/ecam/nep084
20. Khuda-Bukhsh AR, Pathak S. Homeopathic drug discovery: theory update and methodological aspect. Expert Opin. Drug Discov 2008; 3: 979-990.
21. Lewin B. Genes VIII. 2004. Pearson Prentice Hall, Pearson Education International, New Jersy, USA, 2004.
22. Lo SY, Bonavida B. A book containing a dozen experiments using I E crystals. In: Lo SY, Bonavida B, eds. Proceedings of the First International Symposium on I E water clusters. Singapore: World Scientific; 1998; 81-90.
23. Mallick P, Chakrabarti Mallick J, uha B and Khuda-Bukhsh AR. Ameliorating effect of microdoses of a potentized homeopathic drug, Arsenicum Album, on arsenic-induced toxicity in mice. BMC Comp Alt Med, 2003, 3 : 7., available at : http://www.biomedcentral.com/1472-6882/3/7
24. Markovic N, Andersson PU, Nagard MB, et al. Scattering of water from graphite: Simulations and experiments. Chem Phys 1999; 247: 413.
25. Pathak S, Banerjee A, Paul S, Khuda-Bukhsh AR. Protective potentials of a plant extract (Lycopodium clavatum) on mice chronically fed hepato-carciinogens. Indian J Exp Biol, 2009, Vol. 47, July issue, (In Press).
26. Pathak, S., Das JK, Biswas SJ and Khuda-Bukhsh, AR. Protective potentials of a potentized homeopathic drug, Lycopodium-30, in ameliorating azo dye induced hepatocarcinogenesis in mice. Mol Cell Biochem, 2006, DOI: 10.1007/s11010-005-9065-7.
27. Pathak, S., Bhattacharjee N., Das JK, Chaki Choudhury S, Roy Karmakar S, Banerjee, A., Khuda-Bukhsh AR. Supportive evidence for the anticancerous potential of alternative medicine against hepatocarcinogenesis in mice. Forsch Komplementararmed 2007: 14: 148-156. DOI:10.1159/000103280
28. Rey L . Can low-temperature thermoluminiscence cast light on nature of ultra-high dilutions? Homeopathy 2007; 96: 170-74.
29. Samal S, Geckeler KE. Unexpected solute aggregation in water on dilution. Chem Commun 2001; 2224-2225.
30. Watson JD, Baker TA, Bell SP, Gann A, Levine M, Losick R. Molecular biology of the gene,5 th edition. 2004.
31. Wisniewski ES, Folmer DE, Castleman AW jr. Spectroscopic studies of cluster species. Final Program 383, PHYS Fall 2000 Technical Program, 222 nd ACS National Meeting, Chicago, ILL, 2001; 26-30.