Ferrum phosphoricum D12 Treatment Affects J774A.1 Cell Proliferation, Transcription Levels of Iron Metabolism, Antioxidant Defense, and Inflammation-related Genes

Abstract

Background Ferrum phosphoricum (FP) is prescribed as a homeopathic remedy to treat the early stages of fever and inflammation in cases of colds or flu, muscle fatigue and anemia. We aimed to analyze the molecular mechanisms of action of FP D12 on cell proliferation and mRNA expression of iron metabolism, antioxidant defense and inflammation-related genes in mouse J774A.1 macrophages. Methods Cell proliferation was examined using the MTT test. RT-qPCR analyses were performed to estimate gene expression changes. Relative gene expression levels were calculated using the 2–ΔΔCt method. The effect of treatment using FP D12 tablets was compared with that using placebo tablets (PT). Results FP D12 in low concentrations (0.0125 mg/mL to 0.025 mg/mL) significantly stimulated proliferation of J774A.1 cells by up to 11% (p < 0.01) versus control untreated cells and by up to 40% (p < 0.01) versus PT-treated cells in the respective concentration. FP D12 versus PT induced a significant increase in mRNA expression of ferritin light chain (Ftl1) (by 8-fold, p < 0.01), β-2-microglobulin (B2m) (by 2.5-fold, p < 0.05) and iron-responsive element binding protein 2 (Ireb2) (by 4-fold, p < 0.05), and induced a slight decrease in myosin IE (Myo1e) mRNA expression levels (by 0.4-fold, p < 0.01) in macrophages. A highly significant (r2 = 0.99, p < 0.05) correlation was observed between Ireb2 and B2m transcription levels. Significant stimulation of antioxidant enzyme Gpx-1 (by 1.27-fold, p < 0.01) in cells by 0.025 mg/mL FP D12, but a slight decrease (by 0.12-fold, p < 0.05) in 0.0125 mg/mL-treated cells, was observed. A significant increase in the gene expression of IL-1β (by 3.5-fold, р < 0.05) in macrophages was also detected. Conclusion Ferrum phosphoricum in D12 dilution potentially exhibits iron retention, antioxidant and immunomodulation activities, possibly by modulating transcription levels of related genes in non-stimulated mouse macrophages.

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“…And Prof Oskan Tasinov (Bulgaria) described how Ferrum phosphoricum D12 (commonly used in acute infectious inflammatory states) alters the gene expression patterns of activated macrophages in vitro. Prof Tasinov's work was a follow-on project from a study presented in HRI London 2019 11 and forms a growing body of evidence that formally assesses the bioactivity of homeopathic tissue salts, which are a particular type of homeopathic medicine available over-the-counter that have so far been under-represented in the research literature.…”

Acute and Sub-acute Oral Toxicity Assessment of Ferrum phosphoricum in Rats

“…9 FP is also known as "cell salt" due to its oxygen-carrying capacity and related anti-inflammatory action and is used as a firstaid remedy. 10 According to the literature, it has been reported that FP has a potential effect on anemia by improving hemoglobin levels and providing strength to blood vessels. 11 Tasinov et al reported in 2022 that FP at high dilution has iron retention, antioxidant and immunomodulatory activities in mouse macrophages, possibly by modulating transcription levels of related genes.…”

“…10 According to the literature, it has been reported that FP has a potential effect on anemia by improving hemoglobin levels and providing strength to blood vessels. 11 Tasinov et al reported in 2022 that FP at high dilution has iron retention, antioxidant and immunomodulatory activities in mouse macrophages, possibly by modulating transcription levels of related genes. 11 FP has also been found to inhibit the production of free oxygen radicals in human neutrophils.…”

“…11 Tasinov et al reported in 2022 that FP at high dilution has iron retention, antioxidant and immunomodulatory activities in mouse macrophages, possibly by modulating transcription levels of related genes. 11 FP has also been found to inhibit the production of free oxygen radicals in human neutrophils. 12 However, data related to its safety profile at higher dilutions are unavailable.…”

Refining Research in Homeopathy

“…The ongoing pandemic also features in another of our papers: a study at a service sector company in Brazil, which observed a lower incidence of COVID-19 in employees who received a homeopathic medicine for disease prevention compared with those who did not receive the medication. 6 This issue of the journal is completed by two articles with an emphasis on in vitro research, 7 8 together with a clinical case report of individualized homeopathic treatment for co-morbid warts and vitiligo, 9 and a commentary article on the well-known ‘EPI3’ study in France, identifying that program of work as methodologically refined and fit for purpose. 10…”