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EFFECT OF FERRUM PHOSPHORICUM D12 (IRON PHOSPHATE) ON GENE EXPRESSION IN J744A.1 MACROPHAGE CELL LINE

Deyana Vankova, Oskan Tasinov, Milena Pasheva, Desislava Ivanova, Diana Ivanova, Yoana Kiselova

Abstract

Introduction: Regardless of their extensive application and the huge amount of empirical data, scientific research on biological activities of homeopathic remedies is still missing. Ferrum phosphoricum D12 (iron phosphate) (FP) Schussler salt is applied in the initial phase of infections and inflammations, fever, as well as fresh wounds, cuts, scratches, and first-degree burns.

Aim: The aim of the present study was to examine the effect of FP-containing tablets on cell viability and the expression of genes associated with antioxidant protection and inflammatory status in J744A.1 macrophage cells.

Materials and Methods: Commercially available PF-containing tablets and placebo tablets, provided by the manufacturer, were used for cell treatment. Tablets were dissolved in nutrient medium to concentrations from 0.0125 mg/mL to 10 mg/mL. MTT reduction assay was performed to assess cell viability. Expression of antioxidant defense enzymes glutamine-cysteine ligase (GCL) and glutathione peroxidase (GPx) and the inflammatory cytokines interleukin 1 beta (IL-1β), interleukin 6 (IL-6) and TNF-alpha, and the phagocytic enzyme NADPH oxidase was measured using real-time qPCR.

Results: FP–containing tablets of 0.0125 mg/mL and 0.025 mg/mL exerted significant proliferative activity in the medium. Concentrations from 2.5 mg/mL to 10 mg/mL appeared to have increasing cytotoxic effect up to 30%. Treatment of the cells with FP tablets had no significant effect on GPx expression as compared to non-treated cells. However, when compared to placebo control, 2.5 mg/mL and 0.025 mg/mL concentrations were established to increase the expression by 24% and 27%, respectively. Expression of IL-1β gene, interestingly, was significantly affected by placebo 0.0125 mg/mL tablets (67% decrease, p<0.05) when compared to non-treated control. At the same time, the slight nonsignificant stimulatory effect of the 0.0125 mg/mL FP tablets resulted in a 3.5 fold difference (p<0.05) to its respective placebo ones. Obviously, placebo tablets content should be taken into account when evaluating the effects of Schussler salts incorporated into the commercial drug forms. No significant changes in other genes expression levels were established.

Conclusion: This pilot study showed that even in very high dilutions FP preparations may exert biological activity in cell cultures.

Acknowledgements: This study was supported by Alpen Pharma Ltd





DOI: http://dx.doi.org/10.14748/ssp.v4i1.3955

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About The Authors

Deyana Vankova

Oskan Tasinov

Milena Pasheva

Desislava Ivanova

Diana Ivanova

Yoana Kiselova

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