Veronica Elisa Pimenta Vicentini

Assessment of two medicinal plants, Psidium guajava L. and Achillea millefolium L., in in vitro and in vivo assays

The use of medicinal plants by the general population is an old and still widespread practice, which makes studies of their genotoxicity essential. Psidium guajava L. and Achillea millefolium L. are examples of plants commonly used in popular medicine. P. guajava L. is indicated for diarrhea and also as an antiseptic, while A. millefolium L. is indicated as an analgesic, antispasmodic, digestive, diuretic, antiseptic, astringent, emollient, wound healer and hemorrhoid medication. The aim of this study was to determine the effects of the infusions of these two plant species on chromosomes and the cell cycle. Leaves from the plants were used to prepare infusions, in the same manner as teas, but at two different concentrations. Allium cepa L. root-tip cells (P. guajava L. - 2.62 and 26.2 mg/mL, and A. millefolium L. - 3.5 and 35.0 mg/mL) and Wistar rat bone marrow cells (P. guajava L. - 2.62 and 26.2 mg/100g body weight, and A. millefolium L. - 3.5 and 35.0 mg/100g body weight) were used as in vivo plant and animal test systems, respectively. Human peripheral blood lymphocytes (P. guajava L. - 0.262 and 2.62 mg/mL culture medium, and A. millefolium L. - 0.35 and 3.5 mg/mL culture medium) were used as in vitro test system. The P. guajava L. infusion at the higher concentration caused a statistically significant inhibition of cellular division in the onion root-tip cells, not observed in onion root-tip cells treated with A. millefolium L. No statistically significant alterations were found, as compared to untreated controls, in either the cell cycle or the number of chromosome alterations, after treatments with either plant, in rat cells or in cultured human lymphocytes. These results regarding the cytotoxicity and mutagenicity of these plants provide valuable information about the safety of using them as therapeutic agents.

Effects of Maytenus ilicifolia Mart. and Bauhinia candicans Benth infusions on onion root-tip and rat bone-marrow cells

Medicinal plants are widely used to treat various diseases, and in Brazil the plants Maytenus ilicifolia Mart. and Bauhinia candicans Benth are commonly used in popular medicine. However, there are a large number of compounds in plants which can produce alterations in genetic material, and this study was conducted to investigate any possible mutagenic and cytotoxic effects that M. ilicifolia and B. candicans infusions may have on the cell cycle and chromosomes. Infusions were prepared with in natura leaves to give two concentrations of infusions, one at the concentration normally used by the population in general and the other at 10 times this value (i.e. 3.5 and 35 mg/mL for M. ilicifolia and 0.465 and 4.65 mg/mL for B. candicans). Onion (Allium cepa L.) root-tip cells (RTC) and Wistar rat bone-marrow cells (BMC) were used as test systems in in vivo assays. The M. ilicifolia infusions at both concentrations, and the B. candicans infusion at the lower concentration, had no statistically significant depressive mitotic effect on RTC. A statistically significant depressive mitotic effect on RTC was found with the more concentrated (4.65 mg/mL) B. candicans infusion as compared with a negative control. In BMC, infusions of B. candicans and M. ilicifolia produced no statistically significant increase in the number of chromosome alterations or rates of cell division as compared to controls. The significance of these findings are discussed in the light of the use of these plants as therapeutic agents.

Investigation of genotoxic and antigenotoxic activities of chlorophylls and chlorophyllin in cultured V79 cells

Chlorophyll and its derivatives are examples of plant compounds (purified and/or extracted) which appear to protect DNA from damage caused by chemical or physical agents, although some studies have identified clastogenic activity of these compounds. This study was carried out to assess the genotoxic activity of chlorophyll-a (Chl-a), -b (Chl-b) and chlorophyllin (Chl) and their antigenotoxic activity against the DNA damage induced by methyl methanesulphonate (MMS) under conditions of simultaneous, pre-, post-treatment, and simultaneous treatment after pre-incubation of the chemical with MMS. The micronucleus (MN) test was used in binucleated cells (induced by cytochalasin-B) of a mammalian cell line (V79). The three concentrations of Chl-a, Chl-b or Chl (0.1375, 0.275, 0.55microM) were not genotoxic and the genotoxic action of MMS (400microM) decreased (74-117%) under all treatment conditions. The results showed that there was no significant difference among the treatment types, the concentration or the nature of chlorophyll used. The data obtained suggest that Chl-a, Chl-b and Chl when associated with the DNA damaging agent, MMS, may protect the DNA by desgenotoxic action and/or by bio-antigenotoxic mechanisms, with the similar efficiency.

Allium cepa L. as a bioindicator to measure cytotoxicity of surface water of the Quatorze River, located in Francisco Beltrão, Paraná, Brazil

Due to an increase in water consumption in the industrial sector and within the Brazilian population, surface water that receives wastewater from industries, domestic sewage, agricultural industries, and sewage treatment stations can pollute water bodies when not properly treated. The water quality has been linked to catchment characteristics and intensity of agricultural activities. Thus, the aim of this study was to monitor the cytotoxic potential of the water of the Quatorze River, located in the town of Francisco Beltrão, Paraná, Brazil, along its route in the rural area, using the root meristematic cells of Allium cepa L. as a bioindicator. The results showed that the water at points 2, 3, and 4 were not cytotoxic because the rates of A. cepa cell division were unaltered. Point 1 had presented a mitotic index that was statistically larger than the negative control, indicating that this water contained substances with mitogenic capacity, as demonstrated by elevated values in chemical oxygen demand (COD) and biochemical oxygen demand (BOD). However, the mitotic index values decreased along the route of the river (point 1 to point 4), possibly indicating a mechanism of self-purification, despite having received other sources of pollution. Thus, the results of this study show that the water of the Quatorze River should undergo periodic environmental monitoring at different times of the year, including cytotoxicity analysis, to evaluate the principal sources of contamination to maintain the quality of the river water and, consequently, to maintain human health and equilibrium of the entire ecosystem.

Radioprotective effect of the Barbados Cherry ( Malpighia glabra L.) against radiopharmaceutical Iodine-131 in Wistar rats in vivo

BackgroundThe increasing consumption of fruits and vegetables has contributed to the improvement of populational health, due in part, to the abundance of antioxidants in these foods. Antioxidants reduce the level of oxidative damage to DNA caused by free radicals and ionizing radiation, including the radioisotope iodine-131 (131I). This isotope is used for the diagnosis and treatment of thyroid injuries, such as hyperthyroidism and cancer.MethodsThis study aimed to evaluate the radioprotective and cytotoxic activity of acute and subchronic treatments with Barbados Cherry (BC) (Malpighia glabra L.) fruit juice (5 mg), which is rich in potent antioxidants such as vitamin C, phenols, carotenoids, anthocyanins and yellow flavonoids and its activity against the mutagenic activity of the therapeutic dose of 25 μCi of radioiodine for hyperthyroidism. The test system used was the bone marrow cells of Wistar rats (Rattus norvegicus) that were treated in vivo by gavage.ResultsBC showed radioprotective activity in acute treatments, which is most likely due to the joint action of its antioxidant components. In subchronic treatments, the continuous treatment presented an effective radioprotective activity, which was significantly different from treatment with the radiopharmaceutical only. Treatment with BC prior to (PRE) and simultaneous with (SIM) ionizing radiation decreased the number of induced chromosomal alterations, while post-treatment produced no protective effect. In addition, BC exhibited no cytotoxic activity.ConclusionsThese data serve as evidence that BC can be used as a preventive health measure to improve public health quality by countering the action of inevitable exposure to mutagens, such as 131I.

Antimutagenic Effect of Medicinal Plants Achillea millefolium and Bauhinia forficata In Vivo

The investigation of traditionally used medicinal plants is valuable both as a source of potential chemotherapeutic drugs and as a measure of safety for the continued use of these medicinal plants. Achillea millefolium L. (AM) is an ancient remedial herb native to Europe that is used to treat wounds, gastrointestinal and hepatobiliary disorders, inflammation, headaches, and pain. Bauhinia forficata Link (BF), an Asiatic plant, is one of the most commonly used plants in folk medicine against diabetes. The aim of this study was to evaluate the cytotoxic and antimutagenic potential of aqueous extracts of AM and BF on bone marrow cells of Wistar rats treated in vivo. These plant extracts possess considerable antioxidant activity due to the presence of flavonoids and phenolic compounds. These compounds were determinants to noncytotoxic and antimutagenic/protective action of these plants, that reduced statistically the percentage of chromosomal alterations induced by the chemotherapeutic agent cyclophosphamide in simultaneous (AM, 68%; BF, 91%), pre- (AM, 68%; BF, 71%), and post-treatment (AM, 67%; BF, 95%). Therefore, the results of this study indicate that extracts of A. millefolium and B. forficata have antimutagenic potential and that their consumption can benefit the health of those using them as an alternative therapy.

Cytotoxicity and mutagenicity of cola and grape flavored soft drinks in bone marrow cells of rodents

Devido ao grande consumo de refrigerantes no Brasil e no mundo nos ultimos anos, e tendo em vista que alguns dos componentes presentes na composicao destes possuem potenciais danosos para os organismos, em especial o humano, o objetivo deste trabalho foi avaliar o potencial citotoxico e mutagenico de uma marca de refrigerante sabor Cola e uma de sabor Uva. Foram utilizadas como sistema-teste as celulas de medula ossea de ratos Wistar, tratados via gavagem com dose unica do refrigerante sabor Cola ou Uva e, em seguida, fornecidos ad libitum (no lugar da agua), por 24 horas. Foi feito um controle negativo, administrando agua, em dose unica, e um controle positivo administrando ciclofosfamida, via intraperitoneal. A analise estatistica mostrou que os refrigerantes sabor Cola e Uva nao foram citotoxicos. Entretanto, o refrigerante sabor Cola foi mutagenico neste sistema-teste e tempo de tratamento. Desta forma, este estudo serve de alerta para o consumo de refrigerantes sabor Cola e indica que estudos subcronicos e cronicos com os refrigerantes devem ser realizados, a fim avaliar os efeitos mutagenicos e citotoxicos dessas substâncias a longo prazo.

Evaluation of the cytotoxicity, mutagenicity and antimutagenicity of a natural antidepressant, Hypericum perforatum L. (St. John's wort), on vegetal and animal test systems.

BackgroundSt. John’s wort (Hypericum perforatum L.) is an herbaceous plant that is native to Europe, West Asia and North Africa and that is recognized and used worldwide for the treatment of mild and moderate depression. It also has been shown to be therapeutic for the treatment of burns, bruises and swelling and can be used for its wound healing, antiviral, antimicrobial, antioxidant, analgesic, hepato-protective and anxiolytic properties. The aim of this study was to evaluate the potential cytotoxic, mutagenic and antimutagenic action of H. Perforatum.MethodsMeristematic cells were used as the test system for Allium cepa L., and bone marrow cells from Rattus norvegicus, ex vivo, were used to calculate the mitotic index and the percentage of chromosomal aberration. Statistical analysis was performed using the chi-square test.ResultsThis medicinal plant had no cytotoxic potential in the vegetal test system evaluated. In the animal test system, none of the acute treatments, including intraperitoneal gavage and subchronic gavage, were cytotoxic or mutagenic. Moreover, this plant presented antimutagenic activity against the clastogenic action of cyclophosphamide, as confirmed in pre-treatment (76% reduction in damage), simultaneous treatment (95%) and post-treatment (97%).ConclusionsThus, the results of this study suggest that the administration of H. perforatum, especially by gavage similar to oral consumption used by humans, is safe and with beneficial antimutagenic potential.

β-(1→3,1→6)-d-glucans produced by Diaporthe sp. endophytes: Purification, chemical characterization and antiproliferative activity against MCF-7 and HepG2-C3A cells.

This study reports the characterization and antiproliferative activity of exopolysaccharides (EPS) produced by submerged cultures of the endophytes Diaporthe sp. JF766998 and Diaporthe sp. JF767007 isolated from the medicinal plant Piper hispidum Sw. Both strains secreted a crude EPS that, upon size exclusion chromatography, showed to contain a heteropolysaccharide (galactose, glucose and mannose) and a high-molecular weight glucan. Data from methylation analysis, FTIR and NMR spectroscopy (1H, COSY, TOCSY and HSQC-DEPT) indicated that the purified glucan consisted of a main chain of glucopyranosyl β-(1→3) linkages substituted at O-6 by glucosyl residues. According to MTT assay, some treatments of both β-glucans have antiproliferative activity against human breast carcinoma (MCF-7) and hepatocellular carcinoma (HepG2-C3A) cells after 24 and 48h of treatment, exhibiting a degree of inhibition ratio that reached the highest values at 400μg/mL: 58.0% (24h) and 74.6% (48h) for MCF-7 cells, and 61.0% (24h) and 83.3% (48h) for HepG2-C3A cells. These results represent the first reports on the characterization and antiproliferative effect of β-glucans from Diaporthe species and also expand the knowledge about bioactive polysaccharides from endophytic sources.

Effect of Processing, Post-Harvest Irradiation, and Production System on the Cytotoxicity and Mutagenicity of Vitis labrusca L. Juices in HTC Cells

The juices of grapes (Vitis labrusca L.) are similar to the fruit itself because the main constituents of the fruit are present in the juice. However, their quality characteristics may be modified by the harsh technological processes used for the production of integral food, such as production systems of raw materials and post-harvest treatment of grapes with ultraviolet (UV) irradiation. Therefore, the present study analyzed juices produced naturally (by liquefying the fruit) or by the technological process of extraction by steam distillation (90°C) of grapes from organic and conventional production systems that were untreated or treated with UV type C (65.6 J/m2 for 10 minutes). Using cultures of Rattus norvegicus hepatoma cells (HTC) in vitro, cytotoxic effects were assayed by the MTT test and by calculating the cytokinesis blocked proliferation index (CBPI), and mutagenic effects were measured by the cytokinesis block micronucleus assay. The results of the MTT assay and the CBPIs indicated that none of the juices were cytotoxic, including those that induced cell proliferation. The results of the micronucleus assay showed that none of the juices were mutagenic. However, the average number of micronuclei was lower in the juices produced from organic grapes, and cell proliferation, soluble acids and phenolic compounds were significantly higher. Compared with the natural juices, the integral juices of conventional grapes showed a higher average number of micronuclei as well as lower stimulation of cell proliferation and lower levels of bioactive compounds. The results demonstrate a beneficial effect of UV-C irradiation of post-harvest grapes in stimulating the synthesis of nutraceutical compounds without generating cytotoxic or mutagenic substances. Taken together, our findings support the consumption of grape juice and the application of food production techniques that enhance its nutritional value and promote its production, marketing and consumption.