General and genetic toxicology studies of Aleurites moluccana (L.) Willd. seeds in vitro and in vivo assays.

ETHNOPHARMACOLOGY RELEVANCE: Aleurites moluccana is popularly used for the diseases like ulcers, fever, headache, asthma, conjunctivitis, gonorrhea, inflammation, hepatitis, and rheumatism. The seed, also known as "noz da Índia", has been popularly consumed for weight loss purposes but reports of toxicity have been associated with its ingestion. In the literature, there are not enough studies to elucidate its toxicology, so evaluating the general and genetic toxicological of A. moluccana seeds can provide data to ensure their intake. AIM OF THE STUDY: The objective of the present study was to elucidate the oral toxicity, mutagenicity, genotoxicity and cytotoxicity of A. moluccana seeds in vitro and in vivo assays. MATERIALS AND METHODS: The chemical composition of the aqueous extract of A. moluccana seeds (AEAMS) was analyzed in relation to phenolic compounds, tannins, flavonoids and fatty acid. For the in vitro assays, the cytotoxic potential was assessed by the MTS assay whereas the mutagenic potential was assessed by the Ames test. For in vivo assays, was conducted an acute oral toxicity study, with "Up-and-Down Procedure" and repeated dose toxicity with "Repeated Dose 28-Day Oral Toxicity". To assess genetic damage, mutagenic potential was assessed by the micronucleus test whereas the polychromatic erythrocyte/normochromatic erythrocyte ratio was obtained with bone marrow cells to determine the cytotoxic potential and genotoxic potential was assessed by the comet assay using peripheral blood cells. RESULTS: AEAMS did not show cytotoxic and mutagenic potential in vitro. No clinical signs of toxicity were observed in animals after the acute oral toxicity test, suggesting that the LD50 of aqueous extract of A. moluccana seeds > 2000 mg/kg in a single dose by intragastric route. However, in toxicity at repeated doses for 28 days, the doses initially established (250; 500 and 750 mg/kg/day by intragastric route) caused mortality in the animals and the reestablished doses (25, 50 and 100 mg/kg/day by intragastric route) showed no changes in parameters or clinical signs attributed to toxicity. Furthermore, AEAMS also did not show mutagenic, genotoxic and cytotoxic potential in vivo. CONCLUSIONS: AEAMS did not show cytotoxic, genotoxic and mutagenic potential in vitro and in vivo. And although the AEAMS has an LD50 > 2000 mg/kg, and does not have physiological, biochemical, hematological, histopathological changes or clinical signs related to toxicity when administered in low concentrations and for a short period, in high concentrations and continued use caused toxicity and mortality in Wistar rats. In order to obtain complementary results, is recommended highly that further mid and long-term toxicological studies are investigated, and in no-rodent specie.

Cytotoxic, genotoxic and mutagenic evaluation of Alibertia edulis (rich.) a. Rich. ex DC: an indigenous species from Brazil.

Tea leaves of Alibertia edulis is popularly used in folk medicine. However, studies on the genotoxicity of this plant are not available. We aimed to investigate the in vivo and in vitro cytotoxic, genotoxic and mutagenic potentials of the aqueous extract of A. edulis leaves (AEAE). Antioxidant assays, the Artemia salina test, MTT in human platelets, micronucleus in bone marrow and comet in peripheral blood were performed. Animals received four different doses of the AEAE by oral gavage for 30 days. Saline and cyclophosphamide were used as controls. The AEAE exhibited a maximal inhibition at 100 and 250 µg/mL, according to the ABTS and DPPH methods, respectively. The A. salina assay showed that the AEAE presented some toxicity at doses of 100, 250 and 500 µg/mL. Through the MTT assay, the AEAE showed no toxic effects on human platelets during the incubation period. The alkaline comet assay showed that all doses of the AEAE were statistically similar to the negative control group since they did not induce any significant increase of the overall number of damaged cells nor the severity of the cell damage. In the micronucleous assay, results demonstrate that the AEAE did not increase the production of micronucleated polychromatic erythrocytes and was statistically similar to the negative control. The four doses of the plant extract did not affect the production of new erythrocytes and were statistically similar to the negative control groups. Furthermore, the AEAE demonstrated no cytotoxicity, genotoxicity and mutagenicity at the doses tested in rats.

Preclinical safety evaluation of the ethanolic extract from Campomanesia pubescens (Mart. ex DC.) O.BERG (guavira) fruits: analysis of genotoxicity and clastogenic effects.

Genotoxicity studies of plants with medicinal and nutritional properties are recommended by international regulatory agencies as part of the risk assessment. Due to their consumption as food, nutraceutical use and ethnopharmacological relevance, Campomanesia pubescens represents one of these plants to be studied. The aim of the present study was to evaluate the genotoxic, cytotoxic potential and clathogenic effects of the ethanolic extract obtained from the pulp of C. pubescens (EEFCP) fruits on rats submitted to experimental genotoxicity models and through the SMART test performed in Drosophila melanogaster. The comet assay and the micronucleus test were performed on peripheral and bone marrow blood, respectively, of Wistar rats orally treated with EEFCP at doses of 125, 250, 500 and 1000 mg per kg per bw for 28 days. In the SMART test, the standard cross between three mutant D. melanogaster strains was used. Larvae were treated with EEFCP at different concentrations and the wings of adult flies were evaluated for the presence/frequency of mutant spots and compared to the negative control group. Phytochemical analysis of EEFCP indicated high levels of flavonoids. The tests performed in rats showed that EEFCP did not present significant genotoxic or clastogenic effects. The biotransformation metabolites of EEFCP did not present genotoxic activity, as demonstrated by the SMART test. Together, all results indicate that, under the experimental conditions used, EEFCP did not reveal any preclinical genetic toxicity. Therefore, the safe consumption can be fomented increasing, consequently, the economic liquidity in the industrial market from the fruits of guavira.

Preclinical safety evaluation of the ethanolic extract from guavira fruits (Campomanesia pubescens (D.C.) O. BERG) in experimental models of acute and short-term toxicity in rats.

Campomanesia pubescens is a fruit plant widely distributed in South America and used by the population for medicinal and nutritional purposes, with important economic and cultural value. This study evaluated the toxic potential of the ethanolic extract from C. pubescens (EEFCP) fruits through acute and short-term toxicity tests. For the acute toxicity test, female rats received a single oral dose of 2000 mg/kg body weight of EEFCP and were observed for 14 days. In the short-term toxicity test, male and female rats received repeated oral doses of 125, 250, 500 or 1000 mg/kg of EEFCP, being treated and observed for 28 days, and after the treatment period, a satellite and satellite control group remained under observation for another 14 days. No mortality, clinical and organ weight alterations were observed, indicating that LD50 is greater than 2000 mg/kg body weight. In addition, the doses tested did not produce significant changes in the behavioral, physiological, hematological or histopathological parameters of animals. These results demonstrate the low acute and short-term toxicity of EEFCP in rats. The data obtained are of great relevance since they provide important information about a plant species of great economic, nutritional and ethnopharmacological value.

Study on the Cytotoxic, Genotoxic and Clastogenic Potential of Attalea phalerata Mart. ex Spreng. Oil Pulp In Vitro and In Vivo Experimental Models.

Attalea phalerata Mart. ex Spreng. (Arecaceae), popularly known as "bacuri", is used in Brazilian folk medicine. Its oil is used orally to relieve pulmonary congestion and joint pain. In topical applications, it is applied as an effective hair tonic and anti-dandruff. The in natura pulp and its nuts are used as food because of its nutritional value. Despite its use in folk medicine, there is a lack of data regarding its in vivo/in vitro cytotoxic/genotoxic and clastogenic effects. Therefore, in this study, we evaluated the cytotoxic, genotoxic and clastogenic effects of Attalea phalerata Mart. ex Spreng. oil (APMO) in vitro and in vivo. For the analysis of cytotoxic potential, the Artemia salina and MTT (3-(4,5-dimethizzol-zyl)-2,5-diphenyltetrazolium bromide) assays were performed. Possible cytotoxic, genotoxic and clastogenic effects of APMO intake were determined by performing the comet and micronucleus assays. Male and female Wistar rats were orally treated with doses of 125, 250, 500 or 1000 mg.kg-1 of the APMO daily for 28 consecutive days (four weeks). The results showed that the APMO did not induce cell death in the experiments of Artemia salina and MTT, indicating that it has no cytotoxicity. The APMO did not cause significant damage to the DNA of the rats in the four doses used when compared to the negative control group (saline + Tween® 80). The APMO did not present any significant increase in micronucleated polychromatic erythrocytes (MNPCEs) for the four tested doses. When compared to the positive control group, all groups (comet and micronucleus tests) were statistically different. These data suggest that the administration of Attalea phalerata Mart oil. ex Spreng does not cause cytotoxicity, genotoxicity and clastogenicity in experimental models in vitro and in vivo following oral administration in this study.

Toxicological analysis and anti-inflammatory effects of essential oil from Piper vicosanum leaves.

This study assessed the anti-inflammatory effects of the essential oil from Piper vicosanum leaves (OPV) and evaluated the toxicological potential of this oil through acute toxicity, genotoxicity and mutagenicity tests. The acute toxicity of OPV was evaluated following oral administration to female rats at a single dose of 2 g/kg b.w. To evaluate the genotoxic and mutagenic potential, male mice were divided into five groups: I: negative control; II: positive control; III: 500 mg/kg of OPV; IV: 1000 mg/kg of OPV; V: 2000 mg/kg of OPV. The anti-inflammatory activity of OPV was evaluated in carrageenan-induced pleurisy and paw edema models in rats. No signs of acute toxicity were observed, indicating that the LD50 of this oil is greater than 2000 mg/kg. In the comet assay, OPV did not increase the frequency or rate of DNA damage in groups treated with any of the doses assessed compared to that in the negative control group. In the micronucleus test, the animals treated did not exhibit any cytotoxic or genotoxic changes in peripheral blood erythrocytes. OPV (100 and 300 mg/kg) significantly reduced edema formation and inhibited leukocyte migration analyzed in the carrageenan-induced edema and pleurisy models. These results show that OPV has anti-inflammatory potential without causing acute toxicity or genotoxicity.