Dynamized Homeopathic Preparations in Cell Culture- a study report in Amala Cancer Research Centre, Kerala

Courtesy : Oxford Journal

Although reports on the efficacy of homeopathic medicines in animal models are limited, there are even fewer reports on the in vitro action of these dynamized preparations. In Amla Cancer Research Centre, Kerala , Ellanzhiyil Surendran Sunila, Ramadasan Kuttan, Korengath Chandran Preethi and Girija Kuttan have evaluated the cytotoxic activity of 30C and 200C potencies of ten dynamized medicines against Dalton's Lymphoma Ascites, Ehrlich's Ascites Carcinoma, lung fibroblast (L929) and Chinese Hamster Ovary (CHO) cell lines and compared activity with their mother tinctures during short-term and long-term cell culture. The effect of dynamized medicines to induce apoptosis was also evaluated and they have studied how dynamized medicines affected genes expressed during apoptosis. Mother tinctures as well as some dynamized medicines showed significant cytotoxicity to cells during short and long-term incubation. Potentiated alcohol control did not produce any cytotoxicity at concentrations studied. The dynamized medicines were found to inhibit CHO cell colony formation and thymidine uptake in L929 cells and those of Thuja, Hydrastis and Carcinosinum were found to induce apoptosis in DLA cells. Moreover, dynamized Carcinosinum was found to induce the expression of p53 while dynamized Thuja produced characteristic laddering pattern in agarose gel electrophoresis of DNA. These results indicate that dynamized medicines possess cytotoxic as well as apoptosis-inducing properties.

Recently, limited investigations on the efficacy of dynamized medicines in animal models as well as clinical trials have reported that potentiated Lycopodium clavatum has protective action against CCl₄-induced liver damage in rats (3) and that chelidonium 30C could ameliorate both p-dimethylaminoazobenzene and azodye-induced hepatocarcinogenesis in mice (4,5). Anti-genotoxic effect of different dynamized medicines has also been reported (6,7): Arsenicum album was found to ameliorate arsenic-induced toxicity in mice as well as in clinical studies and could reduce the elevated antinuclear antibody titer and hematological toxicities (8,9). Homeopathic therapy for asymptomatic HIV carriers has also proven beneficial (10) and recently Rajendran (11) reported homeopathy as a supportive therapy in cancer. Pathak et al. (12) investigated Ruta 6 on regression of human glioma brain cancer cell growth clinically and found that Ruta induces severe telomere erosion in MGRI brain cancer cells but has no effect on B-lymphoid cells and normal lymphocytes. Banerji and Banerji (13) reported that Ruta was effective for intracranial cysticercosis.

Very few investigations have explored the action of dynamized medicine in in vitro cell culture systems. Podophyllum has been shown to inhibit adhesion of neutrophils to serum-coated micro plates (14). Traumeen S, a homeopathic formulation used clinically to relieve trauma and inflammation has been shown to inhibit the production of Interleukin-β, TNF- and Interleukin-8 by human T cells and monocytes in culture (15). Many homeopathic drugs at low potencies were found to potentiate oxidative metabolism in cultured cells (16).

Medicines Used :

From होम्योपैथी नई सोच/ नई दिशायें

Results:

Although the healing potential of homeopathic drugs is widely accepted, the exact mechanism of action is still unclear. In paragraphs 63–69 of Organon, Hahneman describes the mechanism of action through the ‘primary action’ of the medicine (dynamized or not) and the ‘secondary and curative reaction’ of the organism: ‘Every agent that acts upon the vitality, every medicine, deranges more or less the vital force, and causes a certain alteration in the health of the individual for a longer or a shorter period. This is termed primary action. Although a product of the medicinal and vital powers conjointly, it is principally due to the former power. To its action our vital force endeavors to oppose its own energy. This resistant action is a property, is indeed an automatic action of our life-preserving power, which goes by the name of secondary action or counteraction’. We have tried to explain the mechanism of action of the dynamized preparations taking into consideration the original proposition by Samuel Hahnemann and have approached this problem by investigating the action of dynamized drugs in various cultured cells in a systematic scientific manner.

Cytotoxic activity of a drug is often considered a first step towards elucidating its possible use against cancer and all of the drugs selected are being used by homeopathic practioners against cancer. They have  found that in short-term cytotoxicity research, some of the dynamized preparations showed significant cytotoxic actions against cancer cell lines and at times the activity was higher than that of the mother tinctures. For example, Conium at 200C potency was more cytotoxic than its mother tincture and that the cytotoxicity induced by Carcinosinum was higher at 200C than at 30C potency indicating that dynamization induces the cytotoxic potential of these medications. Results were more pronounced during MTT assay in which a longer period of incubation was involved. Many dynamized preparations at potency of 200C inhibited the growth of L929 cells. Clonogenic assay using CHO cells is a standard method to determine growth inhibitory activity of the drugs and we found that dynamized preparations of Thuja, Hydrastis, Carcinosinum and Podophyllum at 200C potency almost completely inhibited the CHO colony formation. As in other cases, Conium 200C was more active than 30C. We have confirmed the cytotoxic potential of dynamized preparations by thymidine uptake, for the marker of the inhibition DNA synthesis. As in the case other experiments, DNA synthesis was significantly inhibited by several dynamized preparations.

Cytotoxicity could be produced in cells either by necrosis or by apoptotic induction. Apoptosis, which is known as programmed cell death is highly regulated by events taking place within the cell and is highly relevant with respect to the destruction and removal of transformed cells from the body. The induction of apoptosis could be an external agent and a cascade of reactions taking place within the cell produces an ultimate cell death. Some of the events via occurring during apoptosis include morphological changes in the cell, production of apoptotic bodies, damage to genetic material and finally induction of proteolytic enzymes, which produces cellular destruction. Apoptosis could be visualized by morphology and DNA laddering. In the present study, dynamized preparations induced apoptosis as observed from their morphology and DNA laddering. Moreover, dynamized preparation of Carcinosinum could induce the p53, which is considered to be a proapoptotic protein and involved in signal transduction pathway.

The mechanism of action of some of the homeopathic drugs has been proposed. Potentiated preparation of Ruta possesses protective action on normal B-lymphoid cells against H₂O₂-induced chromosomal damage (13). Moreover, the telomere erosion was enhanced in cancer cells by treatment with Ruta while normal cells showed no change. Thus, the telomeres that protect individual chromosomes of cancer cells are damaged by Ruta, which may be the mechanism of its therapeutic action in brain cancer (13).

The protective effect of Chelidonium against p-DAB-induced hepatic cancer may occur by the modulating effect of the drug on restoration of damage caused to several gene-regulated phenomena like enzyme activities and chromosomal abnormalities. This gives insight into the mechanism of action, which may be by means of interfering with the process of carcinogenesis by actively modifying actions of oncogenes or by activating tumor suppressor genes (5). Another mechanism of actions of homeopathic drugs may occur through immune modification. Benveniste (17) has shown that human basophils undergo degranulation not only at usual anti-IgE antibody doses but also at extremely high dilutions. Bastide (18)has shown the therapeutic effect of high dilution of –β interferon and thymic hormones on cellular immunity and had good therapeutic effect in immunodepressed patients. Similarly Bentwich et al. (19) revealed that small amounts of antigens are capable of specific antibody response. The role of immunity in the therapeutic efficacy of homeopathic medicines has also been reviewed (20).

There  results indicate that the dynamized preparation initially produces a secondary action on cells that is in line with the original proposition by Hahnemann on the mechanism of action of medicines dynamized or not. However, limited knowledge in this area does not fully explain the mechanism of action of all drugs . More scientific analyses are warranted to elucidate these interesting preparations of ultra dilutions.

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in pdf : http://ecam.oxfordjournals.org/cgi/reprint/6/2/257

in text : http://ecam.oxfordjournals.org/cgi/content/full/6/2/257