Mercurius solubilis: actions on macrophages.

BACKGROUND: Macrophages play central roles in homeostasis as well as host defence in innate and acquired immunity, auto-immunity and immunopathology. Our research group has demonstrated the effects of highly diluted toxic substances in macrophages. AIM: To investigate if highly diluted Mercurius solubilis (Merc sol), can activate or modulate macrophage functions. METHODS: We evaluated the effects of Merc sol in the 6, 12, 30 and 200 centesimal high dilutions (CH) potencies on mice peritoneal macrophages (in vitro and in vivo). Merc sol was added to mice's drinking water for 7 days (in vivo treatment) and animals were euthanised and cells were collected. In vitro treatment was performed on macrophages and bone-marrow cell cultures. RESULTS: Macrophages showed activated morphology, both when Merc sol was added directly to the cell culture and to drinking water. The in vitro experiments showed enhanced morphological activation, increased interferon (IFN)γ release in the supernatant at lower dilutions and interleukin (IL)-4 production at higher dilutions. Increase in nitric oxide and decrease in superoxide (O(2)(-)) and hydrogen peroxide (H(2)O(2)) were also observed. In vivo treatment caused a decrease in O(2)(-) and increase in H(2)O(2) production by macrophages. DISCUSSION: Taken together, the results allow us to conclude that highly diluted Merc sol modulates reactive oxygen species (ROS), reactive nitrogen species (RNS) and cytokine secretion, which are central mediators of the immune system, wound healing and body homeostasis.

Phagocytosis, endosomal/lysosomal system and other cellularaspects of macrophage activation by Canova medication.

Canova is a homeopathic medication with immunomodulatory properties, recommended for diseases where the immune system is depressed. Our research aims to study the activation of mice peritoneal macrophages when submitted to in vivo and in vitro Canova treatment. Morphological parameters and acid phosphatase activity were analyzed using light and transmission electron microscopy. Differential interference contrast microscopy, including serial time acquisition in living cells, was also performed. The results demonstrated a greater spreading ability in Canova treated macrophages, a higher phagocytic activity of non-infective microorganisms (Saccharomyces cerevisiae and Tripanosoma cruzi epimastigotes) and a tendency to lower the phagocytic activity of the infective microorganisms T. cruzi trypomastigotes and Leishmania amazonensis, when compared with control cells. Acid phosphatase activity was analyzed and showed that Canova treatment stimulates an increase of the endosomal/lysosomal system. Treated macrophages that do or do not interact with yeast present a higher number of acid phosphatase marked vesicles compared to control cells. In contrast, the activity of tartrate resistant acid phosphatase (TRAP), is lower in Canova treated macrophages. The net results demonstrate that Canova medication is an effective stimulator of macrophage activity.

Essiac tea: scavenging of reactive oxygen species and effects on DNA damage.

Essiac, a tea reportedly developed by the Ojibwa tribe of Canada and widely publicized as a homeopathic cancer treatment, is prepared from a mixture of four herbs Arctium lappa, Rumex acetosella, Ulmus rubra and Rheum officinale. Each of these herbs has been reported to possess antioxidant and anti-cancer activity. Essiac itself has also been reported to demonstrate anti-cancer activity in vitro, although its effects in vivo are still a matter of debate. We prepared an extract of Essiac tea from a concentration of 25mg/mL and boiled it for 10 min. From this preparation we used concentrations of 5, 10, 25 and 50% to measure Essiac effects. In this study, we examined the effects of Essiac on free radical scavenging and DNA damage in a non-cellular system, as well as the effects Essiac on lipid peroxidation using the RAW 264.7 cell line. We observed, using electron spin resonance, that Essiac effectively scavenged hydroxyl, up to 84% reduction in radical signal at the 50% tea preparation concentration, and superoxide radicals, up to 82% reduction in radical signal also at the 50% tea preparation concentration, as well as prevented hydroxyl radical-induced DNA damage. In addition, Essiac inhibited hydroxyl radical-induced lipid peroxidation by up to 50% at the 50% tea preparation concentration. These data indicate that Essiac tea possesses potent antioxidant and DNA-protective activity, properties that are common to natural anti-cancer agents. This study may help to explain the mechanisms behind the reported anti-cancer effects of Essiac.

Analysis of IL-2, IFN-gamma and TNF-alpha production, alpha5 beta1 integrins and actin filaments distribution in peritoneal mouse macrophages treated with homeopathic medicament.

The newer forms of immune modulatory therapy are aimed at specific cells or cytokines that contribute to the immune response. These forms of immunotherapy have been referred to as 'biological response modifiers'. Our lab was interested in investigating if a homeopathic medicament 'Metodo Canova' (MC), sold in homeopathic drugstores, does enhance immunological system responses acting through macrophages pathway. Mice peritoneal macrophages were cultivated with or without homeopathic medicament for 24 h for alpha5, beta1 and actin filaments distribution analyses through immunolabelling for confocal microscopy. To detect the IL-2, IFN-gamma and TNF-alpha production these cells were cultivated for 48 h with or without medicament, followed by analyses of these cytokines in supernatant culture with ELISA kits. It was observed differences in morphology and molecular distribution (alpha5 and beta1 integrins, actin filaments and Fc receptors) between the groups control and treated with MC. In control group macrophages had the morphology of resident cells and in MC treated group macrophages were more spread, had many cellular projections and a substantial increase in cytoplasmic volume. In addition, macrophages culture with two doses of MC showed that TNF-alpha production decreased when compared with control group.

Canova, a Brazilian medical formulation, alters oxidative metabolism of mice macrophages.

Macrophages play a significant role in the host defence mechanism. When activated they can produce reactive oxygen species (ROS) as well as related reactive nitrogen species (RNS). ROS are produced via NAD(P)H oxidase which catalyzes superoxide (O2-) formation. It is subsequently converted to hydrogen peroxide (H2O2) by either spontaneous or enzyme-mediated dismutation. Nitric oxide synthase (NOS) catalyzes nitric oxide (NO) formation. Canova (CA) is a Brazilian medication produced with homeopathic techniques, composed of Aconitum, Thuya, Bryonia, Arsenicum, Lachesis in distilled water containing less than 1% ethanol. Previous studies demonstrated that CA is neither toxic nor mutagenic and activates macrophages decreasing the tumor necrosis factor-alpha (TNFalpha) production. In this assay we showed that macrophages triggered with Canova increased NAD(P)H oxidase activity as well as that of iNOS, consequently producing ROS and NO respectively. Cytochrome oxidase and peroxisomes activities were inhibited by NO. As NO and O2- are being produced at the same time, formation of peroxynitrite (ONOO-) may be occurring. A potential explanation is provided on how treatment with Canova may enhance immune functions which could be particularly important in the cytotoxic actions of macrophages. CA can be considered as a new adjuvant therapeutic approach to known therapies.

Effect of Atropa belladonna and Echinacea angustifolia in homeopathic dilution on experimental peritonitis.

Atropa belladonna and Echinacea angustifolia have been used in homeopathy as modulators of inflammatory processes, in simple potency or 'accord of potencies', as recommended by homotoxicology. We evaluated their effects on leukocyte migration and macrophage activity induced by experimental peritonitis in vivo. Mice were injected (i.p.) with LPS (1.0mg/kg) and treated (0.3ml/10g/day, s.c.) with different commercial forms of these medicines. Echinacea angustifolia D4--a simple potency preparation--and Belladonna Homaccord, Belladonna Injeel, Belladonna Injeel Forte, Echinacea Injeel and Echinacea Injeel Forte--all in 'accord of potencies'--were tested. The association of A. belladonna and E. angustifolia in 'accord of potencies' produced an increase of polymorphonuclear cell migration (Kruskal-Wallis, P = 0.03) and a decrease of mononuclear cell percentages (Kruskal-Wallis, P < or = 0.04), when compared with control, mainly in preparations containing low potencies. The proportion of degenerate leukocytes was lower in the treated groups, compared to a control group (P < or = 0.05). The treated groups showed increased phagocytosis (P < or = 0.05), mainly in preparations containing high potencies. Our results suggest that A. belladonna and E. angustifolia, when prepared in 'accord of potencies', modulate peritoneal inflammatory reaction and have a cytoprotective action on leukocytes.