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.
INTRODUCTION: Nowadays Gla-rich protein (GRP) is recognized as a novel biomarker playing a pivotal role in the crosstalk between chronic inflammation and vascular calcification. AIM: The aim of this article is to study the link between circulating GRP, cardiovascular pathology, and the degree of arterial calcification evaluated by the coronary arterial calcium score (CACS) in a Bulgarian population sample. MATERIALS AND METHODS: Adult participants (n=81) of both genders were divided into: controls (n=41)-subjects with estimated moderate-to-high risk without known cardiovascular diseases (CVDs) and a combined CVD group (n=40)-patients with paroxysmal or persistent atrial fibrillation in sinus rhythm, and heart failure subjects with preserved ejection fraction. A structured interview was carried out for evaluation of the classical CVD risk factors. CACS was determined by multislice computed tomography. Routine laboratory parameters were extracted from medical records. Serum levels of total GRP, matrix Gla protein, and osteocalcin were estimated by commercial ELISA kits. Standard statistical methods (descriptive statistics, Student's t-test and Spearman's correlation) were applied. Statistical significance was considered at p<0.05. RESULTS: Significantly lower GRP levels were established in patients with coronary calcium compared to those without calcium deposits. Clear tendency for decreased levels of GRP was observed in the combined CVD group vs controls. Circulating GRP significantly correlates with uncarboxylated matrix Gla protein. An association between serum GRP, CRP, and low-density lipoproteins (LDLs) was demonstrated. CONCLUSION: This study adds new information regarding the role of circulating GRP as a new player in calcification inhibition. Our findings illuminate the link between total circulating GRP, CVD pathology, and the degree of coronary calcification.
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