First cytotoxic, genotoxic, and antigenotoxic assessment of Euterpe oleracea fruit oil (açaí) in cultured human cells.

Euterpe oleracea Mart., popularly known as "açaí", is a tropical fruit from the Amazon region where it has considerable economic importance. Açaí has been used as food and for several medicinal purposes. Despite the widespread use of this fruit, there is a lack of data regarding the safety of using this fruit oil exclusively. Therefore, we evaluated the in vitro cytotoxic, genotoxic, and antigenotoxic effects of E. oleracea fruit oil (EOO) in cultured human lymphocytes (non-metabolizing cells) and HepG2 cell line (human hepatoma) (metabolizing cells) by using MTT, comet, and micronucleus assays. A wide range of EOO concentrations was tested with a preliminary MTT assay, which allowed selecting five concentrations for comet and micronucleus assays: 2.5, 10, 100, 500, and 1000 µg/mL. The results showed that none of the EOO tested concentrations presented cytotoxic effects. The genotoxic assessment revealed an absence of significant DNA and chromosome damage in human lymphocytes and HepG2 cells but did not show chemoprotection against the DNA damage induced by methyl methanesulfonate and benzo[a]pyrene, used as DNA-damaging agents.

Evaluation of the genotoxicity of Euterpe oleraceae Mart. (Arecaceae) fruit oil (açaí), in mammalian cells in vivo.

E. oleracea is a tropical plant from the Amazon region, with its fruit used for food, and traditionally, as an antioxidant, anti-inflammatory, hypocholesterolemic, for atherosclerotic disease, and has anticancer properties. The oil of the fruit has antidiarrheic, anti-inflammatory and antinociceptive activities, but without genotoxicity evaluation. Therefore, the aim of this study was to evaluate the genotoxic potential of E. oleracea fruit oil (EOO), in rat cells. Male Wistar rats were treated with EOO by gavage at doses of 30, 100 and 300 mg/kg, for 14 days, within a 24 h interval. The DNA damage in the leukocytes, liver, bone marrow and testicular cells, was assessed by the comet assay, and the clastogenic/aneugenic effects in the bone marrow cells, by the micronucleus test. Our phytochemicals characterization of the EOO showed the presence of vanillic, palmitic, γ-linolenic, linoleic, oleic, cinnamic, caffeic, protocatechuic, ferulic, syringic acids, and flavonoids quercetin and kaempferol rutinoside as the main constituents. Both cytogenetic tests performed showed that EOO presented no significant genotoxic effects in the analyzed cells, at the three tested doses. These results indicate that, under our experimental conditions, E. oleracea fruit oil did not reveal genetic toxicity in rat cells.

Genotoxic evaluation of the antimalarial drugs artemisinin and artesunate in human HepG2 cells and effects on CASP3 and SOD1 gene expressions.

The malaria treatment recommended by the World Health Organization involves medicines derived from artemisinin, an active compound extracted from the plant Artemisia annua, and some of its derivatives, such as artesunate. Considering the lack of data regarding the genotoxic effects of these compounds in human cells, the objective of this study was to evaluate the cytotoxicity and genotoxicity, and expressions of the CASP3 and SOD1 genes in a cultured human hepatocellular liver carcinoma cell line (HepG2 cells) treated with artemisinin and artesunate. We tested concentrations of 2.5, 5, 7.5, 10, and 20 µg/mL of both substances with a resazurin cytotoxicity assay, and the concentrations used in the genotoxicity experiments (2.5, 5, and 10 µg/mL) and gene expression analysis (5 µg/mL) were determined. The results of the comet assay in cells treated with artemisinin and artesunate showed a significant dose-dependent increase (P < 0.001) in the number of cells with DNA damage at all concentrations tested. However, the gene expression analysis revealed no significant change in expression of CASP3 or SOD1. Our data showed that although artemisinin and artesunate exhibited genotoxic effects in cultured HepG2 cells, they did not significantly alter expression of the CASP3 and SOD1 genes at the doses tested.

Safety and efficacy analysis of liposomal insulin-like growth factor-1 in a fluid gel formulation for hair-loss treatment in a hamster model.

Insulin-like growth factor (IGF)-1 has shown some interesting results in studies examining its use as a hair-loss treatment. IGF-1 works by regulating cellular proliferation and migration during the development of hair follicles. Hepatotoxicity and myelotoxicity were evaluated in hamsters (Mesocricetus auratus) after topical application of the liquid gel vehicle (placebo), 1% IGF-1 or 3% IGF-1. No significant difference in the levels of aspartate aminotransferase or alanine aminotransferase was found between the control and treated groups. ELISA did not shown any increase in the plasma level of IGF-1. A haematopoietic niche was found, but it was not associated with myelotoxicity. Efficacy was determined by dermatoscopy analysis of hair density and microscopy analysis of hair diameter, with hair found to be thicker and with more rapid growth in the 3% group than in either the 1% group or the control group. These results strongly suggest that liposomal IGF-1 in a liquid gel formulation is a safe and efficient treatment for hair loss.

Genotoxicity testing of Ambelania occidentalis (Apocynaceae) leaf extract in vivo.

Ambelania occidentalis is routinely used in folk medicine for treating gastrointestinal disorders, even though there have been no safety trials. We evaluated the genotoxic potential of hydro-alcoholic extracts of this plant in mice; induced DNA damage was assessed in peripheral blood leukocytes and micronucleus induction was assessed in polychromatic erythrocytes from bone marrow. The extract was administered by an oral route at single doses of 1000, 1500 and 2000 mg/kg body weight. N-nitroso-N-ethylurea was used as a positive control. The comet assay was performed on peripheral blood leukocytes at 4 and 24 h after treatment, and the micronucleus test was carried out on bone marrow cells collected at 24 and 48 h after treatment. The ratio of polychromatic/normochromatic erythrocytes was scored for cytotoxicity assessment. No increase in the number of micronucleated polychromatic erythrocytes from bone marrow or in leukocyte DNA damage was observed. The hydro-alcoholic extracts of A. occidentalis had no mutagenic or cytotoxic effects in the mouse cells.

The mutagenic potential of Clusia alata (Clusiaceae) extract based on two short-term in vivo assays.

We examined the genotoxic and mutagenic effects of a crude extract of Clusia alata (a potential medicinal plant) on peripheral leukocyte and bone marrow cells of mice, using the comet and chromosome aberration assays. Extracts at doses of 1000, 1500 and 2000 mg/kg were administered by gavage, and a positive control, N-nitroso-N-ethylurea (50 mg/kg) was injected intraperitoneally. Peripheral blood leukocytes were collected 4 and 24 h after the treatments for the comet assay, and bone marrow cells were collected 24 h after the treatments, for the chromosome aberration assay. The comet assay showed that C. alata extract causes an increase in damage to DNA in the peripheral blood leukocytes, but it was significant only with the 2000 mg/kg dose after 24 h; the extract also induced a small but significant increase in the mean number of chromosome aberrations in the bone marrow cells at doses of 1500 and 2000 mg/kg. No evidence of a significant decrease in the mitotic index was observed. Acute consumption of high concentrations of C. alata extract produced some mutagenic effects in bone marrow cells.

Mutagenicity of the Musa paradisiaca (Musaceae) fruit peel extract in mouse peripheral blood cells in vivo.

Plants are a source of many biologically active products and nowadays they are of great interest to the pharmaceutical industry. In the present study, the mutagenic potential of the Musa paradisiaca fruit peel extract was assessed by the single-cell gel electrophoresis (SCGE) and micronucleus assays. Animals were treated orally with three different concentrations of the extract (1000, 1500, and 2000 mg/kg body weight). Peripheral blood cells of Swiss mice were collected 24 h after treatment for the SCGE assay and 48 and 72 h for the micronucleus test. The results showed that the two higher doses of the extract of M. paradisiaca induced statistically significant increases in the average numbers of DNA damage in peripheral blood leukocytes for the two higher doses and a significant increase in the mean of micronucleated polychromatic erythrocytes in the three doses tested. The polychromatic/normochromatic erythrocyte ratio scored in the treated groups was not statistically different from the negative control. The data obtained indicate that fruit peel extract from M. paradisiaca showed mutagenic effect in the peripheral blood cells of Swiss albino mice.

Lack of clastogenic/genotoxic effects of Baccharis dracunculifolia extract on Swiss mouse peripheral blood cells.

Baccharis dracunculifolia De Candole (DC) (Asteraceae) is indigenous throughout southeastern Latin America and is used by local people in traditional medicine. This plant is known to be the source of resin for the highly valued Brazilian green propolis. As no information is available on the safety of high doses of B. dracunculifolia extract, we evaluated the mutagenic potential of high doses of this plant extract in vivo on peripheral blood cells of Swiss mice using the comet assay and the micronucleus test. The extract was administered by gavage at doses of 1000, 1500 and 2000 mg/kg body weight. Peripheral blood cell samples were collected 4 and 24 h after treatment for the comet assay (genotoxicity assay), and at 48 and 72 h for the micronucleus test (clastogenicity assay). The B. dracunculifolia extract was devoid of clastogenic/genotoxic activity at all doses.

In vivo study of the anti-inflammatory effect of Rhus toxicodendron.

BACKGROUND: Homeopathic Rhus toxicodendron (Rhus tox) is used in various inflammatory conditions. We screened its effect compared to succussed ethanol controls and appropriate active controls. METHOD: We initially experimented with Rhus tox 6, 12, 30 and 200 cH, using carrageenan-induced paw oedema in rats. The 6 cH dilution appeared most effective and was used in subsequent assays. We used pre-treatment and single treatment regimes in Wistar rats, and mice. RESULTS: We found significant reductions compared to control in carrageenan-induced paw oedema, vascular permeability, writhing induced by intraperitoneal acetic acid and stress induced gastric lesions. CONCLUSIONS: Rhus tox in homeopathic dilution appears to interfere with inflammatory processes involving histamine, prostaglandins and other inflammatory mediators.

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.

Anti-inflammatory activity of Arnica montana 6cH: preclinical study in animals.

The anti-inflammatory effect of Arnica montana 6cH was evaluated using acute and chronic inflammation models. In the acute, model, carrageenin-induced rat paw oedema, the group treated with Arnica montana 6cH showed 30% inhibition compared to control (P < 0.05). Treatment with Arnica 6cH, 30 min prior to carrageenin, did not produce any inhibition of the inflammatory process. In the chronic model, Nystatin-induced oedema, the group treated 3 days previously with Arnica montana 6cH had reduced inflammation 6 h after the inflammatory agent was applied (P < 0.05). When treatment was given 6 h after Nystatin treatment, there was no significant inhibitory effect. In a model based on histamine-induced increase of vascular permeability, pretreatment with Arnica montana 6cH blocked the action of histamine in increasing vascular permeability.

Action of Causticum in inflammatory models.

The anti-inflammatory effect of Causticum was evaluated using acute and chronic inflammatory models in vivo. The administration of concentrated Causticum solution into the hind paw of rats produced an inflammatory reaction with oedema formation within the first hour, showing that Causticum acts as an oedematogenic agent. Carrageenin induced rat paw oedema was significantly inhibited (P<0.05) in the group treated with Causticum 30cH solution compared to control. Groups treated with potentized Causticum (6cH, 12cH, 30cH and 200cH), showed significant inhibition (P<0.05) of the inflammation pre-induced by carrageenin. However pre-treatment with Causticum 30cH for 6 days (0.5 ml, daily) did not significantly inhibit granulation using an implantation method.

Central properties of the essential oil and the crude ethanol extract from aerial parts of Artemisia annua L.

The present study evaluated the central activity of the essential oil and the ethanolic extract from Artemisia annua L. in animals as a part of a psychopharmacological screening of this plant. The extract was prepared with fresh leaves in ethanol (AEE) and the essential oil (AEO) was obtained by hidrodestilation. The ED(50) and the LD(50) obtained for the essential oil were 470mg/kg (correlation coefficient r=0.97333 and linear regression y=-26.52x+0.158) and 790mg/kg, and for the extract, 450mg/kg (correlation coefficient r=0.99266 and linear regression y=-27.34+0.156) and more than 2g/kg, respectively. The doses increased the latency time to convulsions induced by picrotoxin and pilocarpine but prevented the onset of pentylenotetrazol and strychnine induced seizures. In addition to, the products have caused marked inhibition in the Rota-rod assay. According to the results, the AEO has a high acute toxicity and a possible cholinergic action, and the AEE showed a possible central activity as dopaminergic and cholinergic agents, and did not present a significant acute toxicity. These differences should be due to chemical substances present in each product. These products had no significant effect as an anticonvulsant, while exhibited a strong depressant activity on the CNS.

Atividade antiinflamatória do granulado de Calendula officinalis L. e Matricaria recutita L.

Este trabalho descreve os resultados da atividade antiinflamatória de um granulado composto pelos extratos secos de Calendula officinalis L. e Matricaria recutita L. (GECOMR® - granulado encapsulado de Calendula officinalis e Matricaria recutita). Estas espécies são utilizadas na medicina popular devido às suas atividades antiinflamatória, antiúlcera gástrica e antiespasmódica. A redução do processo edematogênico nos grupos de animais tratados com o GECOMR foi observada para todos agentes inflamatórios testados (Carragenina, dextrana e histamina). Esta associação mostrou-se eficaz sobre os modelos de edema de pata induzidos pelas diferentes substâncias inflamatórias, com as doses de 100 e 250 mg.kg-1. A associação dos extratos hidroalcoólicos destas espécies medicinais não modificou a indicação isolada de cada uma.

This study shows the results of the anti-inflammatory activity of a phytocomplex composed by dry extracts of Calendula officinalis L. and Matricaria recutita L. (GECOMR®). These plants have been used by folk medicine for its anti-inflammatory, antiulcerogenic and antispasmodic activities. The reduction of the edematogenic process in the treated animals with GECOMR was observed for all inflammatory agents tested, (Carrageenan, Dextran and Hystamine). Its association has been efficient in rat paw edema induced by inflammatory substances in 100 and 250 mg.kg-1 doses. This association of the ethanolic extracts did not modify the indication of each plant isolated.

Atividade antiinflamatória de quefir, um probiótico da medicina popular

O quefir é uma bebida fermentada originária do Cáucaso e formada por bactérias ácido-lácticas e leveduras, sendo utilizada por suas propriedades organolépticas e terapêuticas. O presente estudo avaliou a atividade antiinflamatória deste probiótico sobre modelo de indução de tecido granulomatoso e contorções abdominais induzidas por ácido acético, in vivo. Os resultados obtidos mostraram que o quefir inibiu a formação do tecido granulomatoso, bem como o número de contorções abdominais causadas por ácido acético, quando comparado ao grupo controle. Desta forma, pode-se sugerir uma possível ação antiinflamatória.

Kefir is a fermented beverage with its origin in Caucasian region, formed by acid-lactic bacteria's and yeasts, and used due to its organoleptic and therapeutic properties. The present study evaluated the possibility of an anti-inflammatory activity for induction of granulomatous tissue in rats and writhing test in mice. The results obtained shown that the kefir has inhibited the formation of the granulomatous tissue, as well as the number of writhings induced by acetic acid as compared to a control group. In this way the authors suggest a possible antiinflammatory action.