Toxicological potential of Aloysia gratissima: Insights from chemical analysis and in vitro studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Aloysia gratissima leaves are popularly used to treat respiratory, digestive, and nervous system disorders. Several studies have been carried out to determine the biological activity of A. gratissima, such as its antibacterial and anti-edematogenic activities, but despite the beneficial uses of A. gratissima, few studies have examined the toxicological profile of this plant. AIM OF THE STUDY: This study aimed to determine the chemical composition, cytotoxic, genotoxic, mutagenic potential, and antioxidant activity of an aqueous extract of A. gratissima leaves (AG-AEL). MATERIAL AND METHODS: The phytochemical constitution of AG-AEL was assessed by colorimetric analyses and High-performance liquid chromatography (HPLC). The inorganic elements were detected by Particle-Induced X-ray Emission (PIXE). The antioxidant, cytotoxicity, genotoxic, and mutagenic activities were evaluated in vitro by Di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH), Sulforhodamine B (SRB) assay, comet assay, and Salmonella/microsome assays. RESULTS: AG-AEL indicated the presence of terpenoids, flavonoids, and phenolic acids. HPLC detected rutin at 2.41 ± 0.33 mg/100 mg. PIXE analysis indicated the presence of Mg, Si, P, S, K, Ca, Mn, and Zn. The 50% inhibitory concentration was 84.17 ± 3.17 µg/mL in the DPPH assay. Genotoxic effects were observed using the Comet assay in neuroblastoma (SH-SY5Y) cells and mutations were observed in TA102 and TA97a strains. The extract showed cytotoxic activities against ovarian (OVCAR-3), glioblastoma (U87MG), and colon (HT-29) cancer cell lines. CONCLUSIONS: In conclusion, AG-AEL increased DNA damage, induced frameshift, and oxidative mutations, and showed cytotoxic activities against different cancer cells. The in vitro toxicological effects observed suggest that this plant preparation should be used with caution, despite its pharmacological potential.

Protective effect of Hovenia dulcis Thunb. leaf extracts against ethanol-induced DNA damage in SH-SY5Y cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Hovenia dulcis Thunb. has been used as a medicinal herb for the treatment of hepatic diseases and alcohol intoxication. AIM OF THE STUDY: The genotoxic effect and the antigenotoxic potential of ethanolic extract of H. dulcis leaves and its methanolic fraction were evaluated against ethanol-induced damages in SH-SY5Y cells. MATERIALS AND METHODS: The phytochemical analysis and antioxidant activity of H. dulcis extracts were also assessed. In addition, a systems biology analysis was performed to investigate the molecular pathway of action of the H. dulcis leaves compounds. RESULTS: The ethanolic extract and its methanolic fraction presented genotoxicity through comet assay at 0.5 and 0.25 mg/mL. On the other hand, both extracts showed protective action against ethanol at all concentrations. Additionally, an NBT assay was performed and demonstrated an ability of the extracts to reduce superoxide anion formation when SH-SY5Y cells were challenged with ethanol. HPLC analysis indicated the presence of quercitrin, isoquercitrin, and rutin. Further, system biology assays indicated a molecular action pathway, where the compounds from the leaves of H. dulcis, in addition to performing free radical scavenging activity, activate PP2A, and may inhibit the apoptosis pathway activated by ethanol-induced oxidative stress. CONCLUSIONS: This work is important to indicate potential antigenotoxic and antioxidant properties of H. dulcis leaves, and its use can be investigated against DNA damage induced by ethanol.

Antimicrobial activity of dimeric acylphloroglucinols isolated from southern Brazilian Hypericum species against to resistant bacterial.

The hexanic extracts of Hypericum austrobrasiliense, H. caprifoliatum, H. denudatum, H. pedersenii and H. polyanthemum, and three isolated dimeric acylphloroglucinols (uliginosin B, japonicine A and hyperbrasilol B) were assayed for their antimicrobial activity against some Gram-positive and Gram-negative bacteria (including resistant strains). These extracts were assayed using the disc diffusion test, and the results indicated that the tested species did non exhibit activity on the Gram-negative strains. Subsequently, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were measured using the broth dilution technique adopted to macrodillution. The most susceptible strains were the MRSA and the S. aureus MLSb. Regarding these pathogens, the better MIC values were obtained with the extracts from H. austrobrasiliense, H. caprifoliatum and H. pedersenii. The acylphloroglucinols uliginosin B and hyperbrasilol B presented the lowest MIC values against Enterococcus faecalis, Staphylococcus aureus, MRSA and S. aureus MLSb resistance.

Antioxidant evaluation of the aqueous extract of hulls of Campsiandra laurifolia in colitis induced by acetic acid in Wistar rats / Avaliação antioxidante do extrato aquoso de cascas de Campsiandra laurifolia na colite induzida por ácido acético em ratos Wistar

Abstract Due to the ethnopharmacological use of Campsiandra laurifolia (Fabaceae), popularly known as Acapurana, to treat wounds and ulcers, associated with the lack of alternative treatments for intestinal inflammations such as ulcerative colitis (UC), the present work sought to characterize its phytochemical and antioxidant activities, and to evaluate remedial action in experimental colitis with acetic acid. Phytochemical analyzes were performed through qualitative and quantitative colorimetric tests of the main secondary metabolites. In the colitismodel, 24male Wistar rats aged±60 days oldwere used, divided into 4 groups: Control (CO) control+aqueous extract of C. laurifolia 50mg/kg (CO+A50); Colitis (CL); and Colitis+aqueous extract of C. laurifolia 50 mg/kg (CL+ A50).Measurement of sphincter anal pressure and histological tests of the large intestine, lipoperoxidation (LPO), enzymeactivity of superoxide dismutase (SOD), and levels of glutathione (GSH)were performed. For statistical analysis, the oxidative stress (OS) results were expressed as means±standard error, adopting a significance level of p < 0.05. The screening indicated the presence of flavonoids, saponins and tannins in the extract, with high levels of phenolic

Resumo Devido ao uso etnofarmacológico de Campsiandra laurifolia (Fabaceae), popularmente conhecida comoAcapurana, para tratar feridas e úlceras, associado à falta dealternativas de tratamentos para as inflamações intestinais como a retocolite ulcerativa (RCU), o presente trabalho buscou caracterizar sua constituição fitoquímica, sua atividade antioxidante, e avaliar sua ação reparadora na colite experimental com ácido acético. As análises fitoquímicas foram realizadas por meio de ensaios colorimétricos qualitativos e quantitativos dos principaismetabólitos secundários.Nomodelo de colite, foramutilizados 24 ratos machos Wistar de±60 dias de idade, divididos em 4 grupos: Controle (CO), controle+ extrato aquoso de C. laurifolia 50mg/kg (CO+A50); Colite (CL); e Colite+extrato aquoso de C. laurifolia (CL+ A50). Foram realizadas aferições da pressão anal esfincteriana e avaliações histológicas do intestino grosso, lipoperoxidação (LPO), atividade da enzima superóxido dismutase (SOD) e níveis da glutationa (GSH). Para a análise estatística, resultados do estresse oxidativo (EO) foram expressos em médias±erro padrão, adotando um nível de significância de p < 0,05. O screening indicou no extrato a presença de flavonoides, saponinas e taninos com altos teores de compostos fenólicos e taninos, relacionando-os a uma elevada capacidade antioxidante. Na análise histológica, o grupo CL apresentou perda das criptas, do edema e do infiltrado inflamatório. O uso do extrato de C. laurifolia reestruturou as criptas, diminuiu o edema e aumentou a pressão anal esfincteriana, com diminuição da LPO, da SOD, e aumento da GSH. Sugere-se que o uso do extrato de C. laurifolia diminui o EO por seu poder antioxidante, conferido pelos compostos fenólicos presentes no extrato.

Myrciaria dubia Juice (camu-camu) Exhibits Analgesic and Antiedematogenic Activities in Mice.

The Myrciaria dubia (Myrtaceae) fruit is traditionally used to treat malnutrition due to its high levels of vitamin C and phenolic compounds. Because of its composition, this plant is very promising in the research of novel natural treatment for pain disorders. This study analyzed the phytochemical profile of M. dubia juice and assessed its antinociceptive and antiedematogenic potential. The phytochemical profile was determined through high-performance liquid chromatography (HPLC), the oral antinociceptive effect of M. dubia 50% juice (Md50) was evaluated by formalin, hot plate and Complete Freund's Adjuvant tests and the antiedematogenic activity by paw edema. HPLC revealed the presence of ascorbic acid, rutin, and ellagic acid as major compounds. Md50 showed an antinociceptive effect in the acute and chronic phases of the formalin test. In the hot plate test, Md50 also induced an antinociceptive effect of 0.5 up to 6 h, showing antinociceptive and antiedematogenic potential without changing the spontaneous locomotion of animals. All protocols were submitted and approved by the Ethics Committee for use of Animals of the Lutheran University of Brazil (protocol No. 2013-30P).

In vivo and in vitro toxicological evaluations of aqueous extract from Cecropia pachystachya leaves.

CECROPIA PACHYSTACHYA: leaves are popularly used to treat asthma and diabetes. Despite the widespread consumption of this plant, there are few scientific studies regarding its toxicological potential. In order to conduct a thorough study concerning the potential adverse effects, the aim of this study was to assess acute and subacute toxicity tests of crude aqueous extract from C. pachystachya leaves (CAE-Cp) using in vivomodel, as well as in vitro cytotoxicity, genotoxicity and antioxidant activity. In addition, genotoxicity, and cytotoxicity of chlorogenic acid (CGA) and cytotoxicity of isoorientin (ISOO) were also evaluated. The antioxidant activity was verified by DPPH, cytotoxicity using sulforhodamine B (SRB) assay and genotoxicity by comet assay on V79 cells. The phytochemical analysis of CAE-Cp detected flavonoids and tannins, CGA and ISOO as the major compounds utilizing HPLC. The total flavonoid content (6.52 mg/g EQ) and antioxidant activity (EC50 = 62.15 µg/ml) of CAE-Cp were determined. In vitro evaluations with CAE-Cp showed genotoxic effects at 0.31 to 2.5 mg/ml and an expressive cytotoxicity on HT-29 (IC50 = 4.43 µg/ml) cells. CGA was genotoxic against V79 cells at 0.07 mg/ml and cytotoxic against to HT-29 (IC50 = 71.70 µg/ml), OVCAR-3 (IC50 = 80.07 µg/ml), MCF-7 (IC50 = 45.58 µg/ml) and, NCI-H460 (IC50 = 71.89 µg/ml) cancer cell lines. Wistar rats treated with a single dose (2,000 mg/kg) CAE-Cp decreased hemoglobin levels after 14 days, although no significant toxicity was observed in animals after 28 days. In view of the in vitro cytotoxicity and genotoxicity detected, further studies are necessary to establish the safe use of CAE-Cp.

Anti-hyperlipidemic effects of Campomanesia xanthocarpa aqueous extract and its modulation on oxidative stress and genomic instability in Wistar rats.

The use of natural products from herbs may be a therapeutic option in dyslipidemia treatment. Campomanesia xanthocarpa (Mart.) O. Berg (Myrtaceae) leaves have been used to decrease cholesterol levels. However, studies to determine activities of this plant on triglycerides metabolism have received little attention. The aim of this study was to examine anti-hyperlipidemic effects of a C. xanthocarpa aqueous leaf extract (CxAE) and assess protective actions against oxidative stress and DNA damage. The tyloxapol-induced hyperlipidemia model was used in Wistar rats. Rats were treated orally with CxAE either 250 or 500 mg/kg/day for 7 days prior to tyloxapol administration. Biochemical parameters, oxidative stress levels, and genomic instability were assessed in several tissues. CxAE decreased cholesterol and triglyceride levels in serum and hepatic and renal DNA damage in tyloxapol-treated rats. There was no marked effect on the micronucleus frequency in bone marrow. The extract increased catalase activity and decreased glutathione S-transferase activity in kidney tissue. CxAE showed anti-hyperlipidemic effects, improved oxidative parameters, and protected DNA against damage induced by tyloxapol-induced hyperlipidemia, suggesting C. xanthocarpa leaves may be useful in preventing dyslipidemias.Abbreviations: ALP: Alkaline phosphatase; ALT: Aspartate aminotransferase; ANOVA: Analysis of variance; AST: Aspartate aminotransferase; Ator: Atorvastatin; CAT: Catalase; Chol: Cholesterol; CxAE: Campomanesia xanthocarpa aqueous extract; GST: Glutathione S-transferase; HDL: High density cholesterol; i.p.: Intraperitoneal; NCE: Normochromatic erythrocyte; PBS: Phosphate buffer solution; PCE: Polychromatic erythrocyte; ROS: Reactive oxygen species; SD: Standard deviation; SOD: Superoxide dismutase; T: Tyloxapol; TBARS: Thiobarbituric acid reacting substances; TG: Triglyceride.

Antigenotoxic and antimutagenic effects of Myrciaria dubia juice in mice submitted to ethanol 28-day treatment.

Myrciaria dubia is a native plant from the Amazon region which produces red-purplish fruit rich in antioxidant compounds such as ascorbic acid, carotenoids, and phenolic. M. dubia fruit is used to prepare juices considered to possess high nutritional content providing health benefits. The aim of this study was to examine the ability of M. dubia juice to protect DNA against genomic instability induced by sub-acute ethanol consumption attributed to oxidative stress. Mice were treated for 28 days with juice at 25% and 50% diluted in distilled water or with the diluted combination juice plus ethanol (5 g/kg). The genotoxic/antigenotoxic and mutagenic/antimutagenic effects were assessed using comet assay in blood, liver, and kidney and micronucleus (MN) test with bone marrow. In addition, the mutagenicity was also evaluated using Salmonella/microsome assay. Phytochemical compounds were determined using HPLC/PDA/MS/MS. The juice did not induce genotoxic effects in blood, kidney, and liver cells at both doses. In combination with ethanol, the juice reduced the alcohol-mediated DNA damage in all tissues analyzed. Further, the juice did not produce mutagenic effects and decreased mutagenicity induced by ethanol in the bone marrow. The anthocyanins were major compounds detected by HPLC/PDA/MS/MS, which modulated genotoxic and mutagenic effects initiated by ethanol and at least in part appeared responsible for the observed antigenotoxic and antimutagenic effects of M. dubia juice.

Toxicological aspects of Campomanesia xanthocarpa Berg. associated with its phytochemical profile.

Campomanesia xanthocarpa leaves are used as tea to treat diarrhea, inflammation, and hypercholesterolemia. Some pharmacological studies noted its beneficial uses of C. xanthocarpa; however, few investigations examined the toxicological profile of this plant. The aim of this study was to determine the chemical composition, genotoxic, and mutagenic potential of an aqueous extract of C. xanthocarpa leaves (CxAE), and potential protective effects against oxidative damage. Phytochemical constituents were determined using HPLC, and antioxidant effect in vitro was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical assay. Genotoxic effects and chromosomic mutations were assessed using comet assay and micronucleus (MN) test in Wistar rats treated with CxAE at 250, 500 or 1000 mg/kg for 7 consecutive days. Lipid peroxidation and antioxidant enzyme activities were measured in several tissues. CxAE induced mutations in TA98, TA97a, and TA102 strains. However, in the presence of metabolic activation, data were negative for all strains tested. Lack of mutagenicity was also observed in the MN test. This extract did not induce DNA damage, except when the highest concentration was used. DNA oxidative damage induced by hydrogen peroxide (H2O2) decreased in blood after treatment with CxAE. Lipid peroxidation levels were reduced while superoxide dismutase (SOD) activity increased in kidneys. The inhibitory concentration of CxAE required to lower DPPH levels to 50% was 38.47 ± 2.06 µg/ml. In conclusion, frameshift and oxidative mutations were observed only in the absence of metabolic activation which may be attributed to the presence of flavonoids such as quercetin. It is of interest that CxAE also showed protective effects against DNA oxidative damage associated with presence of ellagic acid, a phenolic acid with antioxidant activities. CxAE did not induce in vivo mutagenicity, suggesting that this extract poses a low toxic hazard over the short term.

Nutritional composition of Eragrostis teff and its association with the observed antimutagenic effects.

Eragrostis teff is an Ethiopian native grass plant (Poaceae or Gramineae family) whose importance as a crop grain has increased in recent years. The aim of this study is to analyze the nutritional composition of its seeds and the mutagenic/antimutagenic activity of the hydroalcoholic extract of the seed flour. Chemical elements (colloquially known as minerals) were determined using Particle-Induced X-ray Emission (PIXE) and Flame Atomic Absorption Spectroscopy (FAAS), while the content of amino acids (aminogram) and fatty acids (profile of fatty acids) were quantified by HPLC. Mutagenic activities were tested using Salmonella/microsome assay. Mutagens doxorubicin, 4-nitroquinolin N-oxide, methylmethanosulphonate, and aflatoxin B-1 were used in Salmonella typhimurium TA98 and TA100 strains to assess antimutagenic activities. The major elements observed were K, P, S, Mg, and Ca. Almost all essential amino acids were observed and the predominance of unsaturated fatty acids in the total oil content of 2.72% (w/w) is also noted, including the two essential fatty acids alpha-linolenic acid (an omega-3 fatty acid) and linoleic acid (an omega-6 fatty acid). Hydroalcoholic extract of E. teff seed flour showed antimutagenic activity, protecting against frameshift and base pair substitution mutations. These findings provide valuable information for further development of healthier foods that can be produced with increasing yields and minimal environmental impact.

Genotoxic and chemopreventive assessment of Cynara scolymus L. aqueous extract in a human-derived liver cell line.

Cynara scolymus L., popularly known as artichoke, is consumed as food and used as tea infusions for pharmacological purposes to treat liver dysfunctions and other conditions. Scientific data on the safety and protective effect of artichoke in human-derived liver cells is missing. This study investigated the genotoxic and modulatory effect of a liophilized extract suspended in water of C. scolymus L. leaves. Four extract concentrations (0.62, 1.25, 2.5 and 5.0 mg/mL) were evaluated using the comet assay on human hepatocyte cultures, HepG2 cells. Genotoxicity was assessed after two treatment periods, 1 and 24 h. Antigenotoxicity was evaluated against oxidative lesions induced by hydrogen peroxide in pre-, simultaneous and post-treatment protocols. Artichoke leaves aqueous extract induced genotoxic effects in HepG2 cells after 1- and 24-h treatments. In turn, extract concentrations of 0.62, 1.25 and 2.5 mg/mL, exhibited a protective effect in pretreatment, compared to hydrogen peroxide alone. However, in simultaneous and post-treatment protocols, only the lowest concentration reduced the frequency of DNA damage induced by hydrogen peroxide. In addition, in the simultaneous treatment protocol, the highest artichoke extract concentration increased hydrogen peroxide genotoxicity. It can be concluded that artichoke is genotoxic, in vitro, to HepG2 cells, but can also modulate hydrogen peroxide DNA damage.

Protective effects of acerola juice on genotoxicity induced by iron in vivo.

Metal ions such as iron can induce DNA damage by inducing reactive oxygen species (ROS) and oxidative stress. Vitamin C is one of the most widely consumed antioxidants worldwide, present in many fruits and vegetables, especially inMalpighia glabra L., popularly known as acerola, native to Brazil. Acerola is considered a functional fruit due to its high antioxidant properties and phenolic contents, and therefore is consumed to prevent diseases or as adjuvant in treatment strategies. Here, the influence of ripe and unripe acerola juices on iron genotoxicity was analyzed in vivo using the comet assay and micronucleus test. The comet assay results showed that acerola juice exerted no genotoxic or antigenotoxic activity. Neither ripe nor unripe acerola juices were mutagenic to animals treated with juices, in micronucleus test. However, when compared to iron group, the pre-treatment with acerola juices exerted antimutagenic activity, decreasing significantly micronucleus mean values in bone marrow. Stage of ripeness did not influence the interaction of acerola compounds with DNA, and both ripe and unripe acerola juices exerted protective effect over DNA damage generated by iron.

Evaluation of Safety of Arrabidaea chica Verlot (Bignoniaceae), a Plant with Healing Properties.

Arrabidaea chica Verlot (Bignoniaceae) has been used as a medicinal herb to treat anemia, hemorrhage, inflammation, intestinal colic, hepatitis, and skin infections in the Brazilian Amazon region. Studies have demonstrated the healing properties of extracts obtained from A. chica leaves, which contain anthocyanins and flavonoids. However, few investigations have assessed the safe use of this plant species. In this study, mutagenic and genotoxic effects of a crude aqueous extract, a butanolic fraction, and aqueous waste from A. chica leaves were evaluated using the Salmonella/microsome assay in TA98, TA97a, TA100, TA102, and TA1535 strains and the alkaline comet assay in Chinese hamster ovary (CHO) cell culture with and without metabolic activation. The crude aqueous extract, butanolic fraction, and aqueous waste were not mutagenic in any of the Salmonella typhimurium strains tested, and showed negative responses for genotoxicity in CHO cells. High-performance liquid chromatography (HPLC) analysis indicated the presence of phenolic acids and flavonoids such as rutin and luteolin. The lack of mutagenic/genotoxic effects might be due to phytochemical composition with high concentrations of known anti-inflammatory compounds. Thus, the crude aqueous extract, butanolic fraction, and aqueous waste from A. chica leaves do not appear to pose short-term genotoxic risks.

Corrective effects of acerola (Malpighia emarginata DC.) juice intake on biochemical and genotoxical parameters in mice fed on a high-fat diet.

Acerola contains high levels of vitamin C and rutin and shows the corresponding antioxidant properties. Oxidative stress on the other hand is an important factor in the development of obesity. In this study, we investigated the biochemical and antigenotoxic effects of acerola juice in different stages of maturity (unripe, ripe and industrial) and its main pharmacologically active components vitamin C and rutin, when given as food supplements to obese mice. Initial HPLC analyses confirmed that all types of acerola juice contained high levels of vitamin C and rutin. DPPH tests quantified the antioxidant properties of these juices and revealed higher antioxidant potentials compared to pure vitamin C and rutin. In an animal test series, groups of male mice were fed on a standard (STA) or a cafeteria (CAF) diet for 13 weeks. The latter consisted of a variety of supermarket products, rich in sugar and fat. This CAF diet increased the feed efficiency, but also induced glucose intolerance and DNA damage, which was established by comet assays and micronucleus tests. Subsequently, CAF mice were given additional diet supplements (acerola juice, vitamin C or rutin) for one month and the effects on bone marrow, peripheral blood, liver, kidney, and brain were examined. The results indicated that food supplementation with ripe or industrial acerola juice led to a partial reversal of the diet-induced DNA damage in the blood, kidney, liver and bone marrow. For unripe acerola juice food supplementation, beneficial effects were observed in blood, kidney and bone marrow. Food supplementation with vitamin C led to decreased DNA damage in kidney and liver, whereas rutin supplementation led to decreased DNA damage in all tissue samples observed. These results suggest that acerola juice helps to reduce oxidative stress and may decrease genotoxicity under obesogenic conditions.

Protective activity of Cynara scolymus L. leaf extract against chemically induced complex genomic alterations in CHO cells.

Cynara scolymus L., popularly known as artichoke, has been widely used in traditional medicine as an herbal medicament for therapeutic purposes. The study aimed at assessing the protective activity of Cynara scolymus leaf extract (LE) against DNA lesions induced by the alkylating agent ethylmethnesulphonate (EMS) in Chinese hamster ovary cells (CHO). The ability of C. scolymus L. LE to modulate the mutagenicity of EMS was examined using the cytokinesis block micronucleus (CBMN) cytome assay in three antigenotoxic protocols, pre- post- and simultaneous treatments. In the pre-treatment, C. scolymus L. LE reduced the frequencies of MNi and NBUDs induced by EMS in the lower concentration. In contrast, at the highest concentration (5 mg/ml) artichoke enhanced the frequency of MNi, potentiating EMS genotoxicity. In the simultaneous treatment only the induction of MNi was repressed by the exposure of cells to C. scolymus L. LE. No modification in genotoxicity was observed in LE post-treatment. The results obtained in this study suggest that lower concentrations of artichoke prevent chemically induced genomic damage in mammalian cells. In this context, the protective activity of C. scolymus L. could be associated to its constitutive antioxidants compounds.

Evaluation of acute and subacute toxicity and mutagenic activity of the aqueous extract of pecan shells [Carya illinoinensis (Wangenh.) K. Koch].

The infusion of pecan shells has been used to prevent and control hypercholesterolemia, diabetes and toxicological diseases. The aim of the present study was to evaluate toxicity and mutagenic effects of pecan shells aqueous extract (PSAE). Wistar rats were treated with a single dose of 300 or 2000 mg/kg of PSAE in the acute toxicity test. For the subacute test, the animals received 10 or 100 mg/kg of PSAE for 28 days. The mutagenicity was evaluated using Salmonella/microsome assay in TA1535, TA1537, TA98, TA100 and TA102 S. typhimurium strains in the presence and absence of metabolic activation (S9 mix) and micronucleus test in bone marrow. HPLC analyses indicated the presence of tannins, flavonoids, gallic and ellagic acids. Except for triglycerides, all treated groups presented normal hematological and biochemical parameters. Lower levels of triglycerides and weight loss were observed in the 100 mg/kg group. Mutagenic activities were not detected in S. typhimurium strains and by the micronucleus test. Based on these results, PSAE was not able to induce chromosomal or point mutations, under the conditions tested. The 100mg/kg dose showed significant antihyperlipidemic action, with no severe toxic effects.

Evaluation of the mutagenicity and genotoxicity of Arrabidaea chica Verlot (Bignoneaceae), an Amazon plant with medicinal properties.

Arrabidaea chica Verlot (Bignoniaceae) is an important folk medicine plant native to the Amazon region and used to treat anemia, hemorrhage, inflammation, intestinal colic, hepatitis, and skin affections. Although studies showed its therapeutic properties, little knowledge regarding genotoxic properties of this plant is available. The aim of this study was to determine the potential mutagenic and genotoxic/antigenotoxic effects of an A. chica chloroformic fraction (Ac-CF) obtained from leaves containing bioactive metabolites. The mutagenic effects were evaluated using the Salmonella mutagenicity assay, with TA98, TA97a, TA100, TA102, and TA1535 strains, with and without metabolic activation. In vivo mutagenic and genotoxic/antigenotoxic effects were investigated using the micronucleus (MN) test in bone marrow and alkaline comet assay in blood and liver after administration of 100, 500, or 1000 mg/kg Ac-CF in CF-1 mice by gavage (once a day for 3 d). In vitro antioxidant potential was evaluated using DPPH and xanthine/hypoxanthine assays. Ac-CF was not mutagenic in any of the Salmonella typhimurium strains tested and showed negative responses for mutagenicity and genotoxicity in mice. Further, Ac-CF displayed antigenotoxic effects by decreasing the oxidative DNA damage induced by hydrogen peroxide by greater than 50% in blood and liver. The antioxidant action detected in the in vitro assays demonstrated IC50 of 0.838 mg/ml in the xanthine/hypoxanthine assay and IC50 of 28.17 µg/ml in the DPPH assay. In conclusion, Ac-CF did not induce mutagenic and genotoxic effects and was able to protect DNA against oxidative damage in vivo, suggesting that this fraction may not pose genetic risks, although further toxicology assays are necessary.

Artichoke induces genetic toxicity in the cytokinesis-block micronucleus (CBMN) cytome assay.

Artichoke leaves are used in traditional medicine as an herbal medicament for the treatment of hepatic related diseases, as well as choleretic and diuretic. The aim of the present study was to evaluate the capacity of Cynara scolymus L. leaves extract (LE) to cause chromosomal instability and cytotoxicity in Chinese hamster ovary cells (CHO) employing the cytokinesis-block micronucleus (CBMN) cytome assay. Cells were treated with four concentrations of C. scolymus for two exposure times: 1h and 24h. Our findings showed that LE did not increase the frequencies of nucleoplasmic bridges (NPBs) and nuclear bud (NBUD). However, all concentrations of the extract produced increments in micronuclei frequencies (MNi) in both exposure times, when compared to the negative control. No significant differences were observed in the nuclear division cytotoxicity index (NDCI), reflecting the absence of cytotoxic effects associated to LE. The results demonstrated the ability of C. scolymus LE to promote chromosomal mutations which are, probably, a result of the pro-oxidant activity of LE constituents such as flavonoids and chlorogenic acids. The data obtained in this study suggests that high concentrations of artichoke can pose a risk associated to its consumption.

Genotoxic and antigenotoxic activity of acerola (Malpighia glabra L.) extract in relation to the geographic origin.

Malpighia glabra L, popularly known as acerola, is considered a functional fruit and therefore is taken to prevent disease or as adjuvant to treatment strategies, since the fruit is an undeniable source of vitamin C, carotenoids, and flavonoids. Acerola is a natural source of vitamin C, flavonoids, and carotenoids. Its chemical composition is affected by genetic uniformity of the orchards and environmental factors. Considering the extensive growth of the culture of acerola in Brazil as well as its widespread use, this study evaluates the genotoxic and antigenotoxic activity of acerola in relation to geographical origin using the comet assay in mice blood cells in vitro. No acerola samples showed potential to induce DNA damage, independently of origin. Also, for antigenotoxicity activity, only the acerola sample from São Paulo reduced DNA damage induced by hydrogen peroxide (by about 56%). The sample from Ceará showed good antioxidant activity by the 2,2-diphenyl-1-picrylhydrazyl assay, in agreement with its higher rutin, quercetin, and vitamin C levels. Additional studies with other treatment regimens are necessary to better understand the impact of the complex mixture of acerola on genomic stability.

Artichoke induces genetic toxicity and decreases ethyl methanesulfonate-related DNA damage in Chinese hamster ovary cells.

Cynara scolymus L. (Asteraceae), popularly known as artichoke, has been widely used in herbal medicine for the treatment of hepatic diseases. The genotoxicity of C. scolymus L. leaf extract (LE) and the ability to modulate the genetic toxicity of the alkylating agent ethyl methanesulfonate (EMS) were assessed using the comet assay on Chinese hamster ovary cells. Genotoxicity was evaluated after 1- and 24-h treatments using four different LE concentrations: 0.62, 1.25, 2.5, and 5.0 mg/mL. Antigenotoxicity was assessed for pretreatment, simultaneous treatment, and post-treatment. All doses used led to a significant increase in the frequency of DNA damage, after exposure for 1 and 24 h. In the antigenotoxicity experiments, LE reduced the frequency of DNA damage induced by EMS in the simultaneous treatment only. However, the lowest dose was more protective than higher concentrations. Flavonoids and phenolic compounds are, probably, the C. scolymus constituents responsible for its genotoxic and antigenotoxic effects.