ABSTRACT
This study aimed to evaluate the modes of action of harringtonine (HT) and homoharringtonine (HHT) alkaloids in cell with wild (HepG2/C3A) and mutant p53 (HuH-7.5). We performed assays for cytotoxicity, genotoxicity, induction of apoptosis, cell cycle phase, and membrane integrity. Obtained data were compared with the relative expression of mRNA of genes related to proliferation, apoptosis, cell cycle control, metabolism of xenobiotics, and reticulum endoplasmic stress. The relative expression of the genes showed an increase in apoptosis-inducing mRNAs, such as TNF and BBC3, as well as a reduction in BCL2 and BAK. The mRNAs of CYP2E1 and CYP2C19 xenobiotic metabolism genes increased in both lineages, while CYP3A4 increased only in the HuH-7.5 lineage. The mRNA expression of endoplasmic reticulum (ER) stress genes (ERN1 and EIF2AK3) was shown to increase in HHT and HT treatments. A similar increase was recorded in the mRNA expression of the TRAF2 gene. The changes observed in this study support the hypothesis that ER stress was more strongly associated with TNF induction, causing cell death by apoptosis in p53 mutant cells. This result with wild and mutant p53 cells may have clinical implications in the use of these compounds.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Carcinoma, Hepatocellular/drug therapy , Harringtonines/pharmacology , Liver Neoplasms/drug therapy , Tumor Suppressor Protein p53/genetics , Apoptosis/drug effects , Carcinoma, Hepatocellular/genetics , Cell Cycle/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Endoplasmic Reticulum Stress/drug effects , Humans , Liver Neoplasms/geneticsABSTRACT
AIMS: Fructooligosaccharides (FOSs) known for their health properties and ß-(2â6)-levan-type FOSs have shown prebiotic and immunomodulatory activities that overcome those of commercial ß-(2â1)-FOSs, but costs do not favour their use. Moreover, FOSs can reach the bloodstream through the diet, and little is known about their direct effect on cells. The aim of this work was to produce high-content FOSs by Bacillus subtilis natto CCT7712 in a bioreactor using commercial sucrose and to evaluate their antiproliferative effects in OVCAR-3 cells. METHODS AND RESULTS: FOS production reached 173·60 g l-1 , 0·2 vvm aeration and uncontrolled pH. Levan-type FOSs, composed of ß-(2 â 6) links and mainly GF3 (6-nystose), were identified using RMN spectroscopy, FT-IR and ESI-MS. FOSs decreased the viability and proliferation of OVCAR-3 cells, and the effects were associated with an increased pro-inflammatory response by the induction of IL-8 and TNF-α, and the repression of ER-ß genes. The metabolic profiles showed disruption of cellular homeostasis that can be associated with a decrease in proliferation. CONCLUSIONS: The high production of levan-type FOSs from B. subtilis natto CCT7712 in a bioreactor was achieved, and they showed antiproliferative potential in OVCAR-3 cells. SIGNIFICANCE AND IMPACT OF THE STUDY: FOS could be a good target for future therapeutic studies and commercial use.
Subject(s)
Bacillus subtilis/metabolism , Cell Proliferation/drug effects , Oligosaccharides/metabolism , Oligosaccharides/pharmacology , Bioreactors , Cell Line, Tumor , Cell Survival/drug effects , Fructans/chemistry , Fructans/metabolism , Fructans/pharmacology , Humans , Oligosaccharides/chemistry , Sucrose/metabolismABSTRACT
The compounds 6-dimethylaminopurine (6-DMAP) and cyclohexamide (CHX) are currently used to stimulate the development of embryos produced by nuclear transfer in the production of cloned mammals with a great deal success. This study investigated the effects of 6-DMAP and CHX in vivo using biological assays to evaluate reproductive performance in females, teratogenesis, and mutagenesis. The results of this study demonstrated that the activating agents of oocyte cytoplasm, 6-DMAP and CHX, altered the reproductive performance of the experimental animals, as well as increased the rate malformations. In addition to these adverse effects, the administration of these compounds in pregnant females resulted in genotoxic and mutagenic toxicity, as determined by comet and micronucleus assays carried out in peripheral blood samples, respectively. Based on these findings and that alterations in DNA are important, morpho-functional teratogenesis and diminished embryonic viability, suggesting that 6-DMAP and CHX, which are utilized during the cloning of mammals, are responsible for the fact that embryos produced by nuclear transfer show low viability after implantation in utero or after birth because of congenital malformations and/or alterations in their DNA.
Subject(s)
Adenine/analogs & derivatives , Cloning, Organism , Cycloheximide/adverse effects , Mutagens/adverse effects , Reproduction/drug effects , Adenine/adverse effects , Animals , Embryo Transfer , Female , Mice , Pregnancy , Reproduction/genetics , Teratogenesis/drug effectsABSTRACT
Polyphenolic compounds present in rosemary were found to have antioxidant properties, anticarcinogenic activity, and to increase the detoxification of pro-carcinogens. The aim of the study was to determine the effect the aqueous extract of rosemary (AER) on mutagenicity induced by methylmethane sulfonate in meristematic cells of Allium cepa, as well as to describe its mode of action. Anti-mutagenicity experiments were carried out with 3 different concentrations of AER, which alone showed no mutagenic effects. In antimutagenicity experiments, AER showed chemopreventive activity in cultured meristematic cells of A. cepa against exposure to methylmethane sulfonate. Additionally, post-treatment and simultaneous treatment using pre-incubation protocols were the most effective. Evaluation of different protocols and the percent reduction in DNA indicated bioantimutagenic as well desmutagenic modes of action for AER. AER may be chemopreventive and antimutagenic.
Subject(s)
Antimutagenic Agents/pharmacology , Meristem/cytology , Mutagens/pharmacology , Onions/cytology , Plant Extracts/pharmacology , Rosmarinus/chemistry , Water/chemistry , Chromosome Aberrations , DNA Damage , Methyl Methanesulfonate/pharmacology , Mitotic IndexABSTRACT
We evaluated the effects of glutamine on clastogenic and genotoxic damage prevention caused by the administration of cisplatin. Forty Swiss mice were divided into 8 experimental groups: G1 and G2, which were control groups; G3, G4, and G5, which were administered [2 doses of glutamine (orally)] separated by a 24-h period (150, 300, and 600 mg/kg, respectively), and a dose of phosphate-buffered saline by intraperitoneal injection; G6, G7, and G8, which were treated in the same manner as the previous groups, but received cisplatin rather than phosphate-buffered saline. The antimutagenicity groups showed damage reduction percentages of 79.05, 80.00, and 94.27% at the time point T1, 53.18, 67.05, and 64.74 at time point T2 for the 150, 300, and 600 mg/kg doses of glutamine, respectively. Antigenotoxic activity was evident for all 3 doses with damage reduction percentages of 115.05, 119.06, and 114.38 for the doses of glutamine of 150, 300, and 600 mg/ kg, respectively. These results suggest that further studies are needed to confirm the clastogenic activity of glutamine. However, our results may lead to rational strategies for supplementation of this antioxidant as an adjuvant in cancer treatment or for preventing genomic lesions.
Subject(s)
Antimutagenic Agents/pharmacology , Antioxidants/pharmacology , Cisplatin/pharmacology , Glutamine/pharmacology , Mutagens/pharmacology , Animals , Antineoplastic Agents/pharmacology , Cisplatin/antagonists & inhibitors , Comet Assay , DNA Damage , Injections, Intraperitoneal , Male , Mice , Micronucleus TestsABSTRACT
The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.
Subject(s)
Adenine/analogs & derivatives , Cloning, Organism/methods , Comet Assay , Cycloheximide/toxicity , Mutagens/toxicity , Adenine/toxicity , Animals , Cell Culture Techniques , Cell Survival/drug effects , Coloring Agents , Cytokinesis/drug effects , Hep G2 Cells/drug effects , Humans , Male , Mammals , Mice , Micronucleus Tests , Mutagenicity Tests , Nuclear Transfer Techniques , Tetrazolium Salts/pharmacology , Thiazoles/pharmacology , Trypan Blue/pharmacologyABSTRACT
The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.
Subject(s)
Animals , Humans , Male , Mice , Adenine/analogs & derivatives , Comet Assay , Cloning, Organism/methods , Cycloheximide/toxicity , Mutagens/toxicity , Adenine/toxicity , Cell Culture Techniques , Coloring Agents , Cell Survival/drug effects , Cytokinesis/drug effects , /drug effects , Mammals , Micronucleus Tests , Mutagenicity Tests , Nuclear Transfer Techniques , Tetrazolium Salts/pharmacology , Thiazoles/pharmacology , Trypan Blue/pharmacologyABSTRACT
Fluoxetine, commonly known as Prozac, is the first representative of the so-called new generation of antidepressants that promise efficacy, with few side effects, against deep depression, nervous bulimia, and anxiety. As there is a growing number of people suffering from anxiety and depression; consequently, the use of fluoxetine is also increasing. Verifying absence of drug effects such as cytotoxicity or mutagenicity is of great importance. Certain vitamins, such as vitamin A (retinol, retinoids) and vitamin C (ascorbic acid) protect and are extremely active against mutagens. We evaluated the cytotoxic and mutagenic activity of fluoxetine, with and without concomitant administration of vitamin A or C, in Allium cepa meristem cells and Wistar rat bone marrow cells. The A. cepa meristem cells showed fluoxetine cytotoxicity; concomitant treatment with vitamin A or C proved non-protective. Treatment of Wistar rats with fluoxetine intraperitoneally or via gavage did not affect cell division or cause clastogenic effects. Vitamin A and C did not affect the cytotoxicity or mutagenicity of fluoxetine in the rat cells.
Subject(s)
Fluoxetine/administration & dosage , Mutagens/administration & dosage , Onions/drug effects , Animals , Ascorbic Acid/administration & dosage , Cell Division/drug effects , Drug Combinations , Fluoxetine/adverse effects , Humans , Mutagenicity Tests , Mutagens/adverse effects , Onions/genetics , Rats , Vitamin A/administration & dosageABSTRACT
This study evaluated the mutagenicity and antimutagenicity of inulin in a chromosomal aberration assay in cultures of the meristematic cells of Allium cepa. The treatments evaluated were as follows: negative control--seed germination in distilled water; positive control--aqueous solution of methyl methanesulfonate (10 µg/mL MMS); mutagenicity--aqueous solutions of inulin (0.015, 0.15, and 1.50 µg/mL); and antimutagenicity--associations between MMS and the different inulin concentrations. The antimutagenicity protocols established were pre-treatment, simultaneous simple, simultaneous with pre-incubation, and post-treatment. The damage reduction percentage (DR%) was 43.56, 27.77, and 55.92% for the pre-treatment; -31.11, 18.51, and 7.03% for the simultaneous simple; 30.43, 19.12, and 21.11% for the simultaneous with pre-incubation; and 64.07, 42.96, and 53.70% for the post-treatment. The results indicated that the most effective treatment for inhibiting damages caused by MMS was the post-treatment, which was followed by the pre-treatment, suggesting activity by bioantimutagenesis and desmutagenesis. The Allium cepa assay was demonstrated to be a good screening test for this type of activity because it is easy to perform, has a low cost, and shows DR% that is comparable to that reported studies that evaluated the prevention of DNA damage in mammals by inulin.
Subject(s)
Antimutagenic Agents/pharmacology , Chromosome Aberrations/drug effects , Inulin/pharmacology , Methyl Methanesulfonate/pharmacology , Mutagens/pharmacology , Onions/drug effects , Cells, Cultured , DNA Damage , Meristem/cytology , Meristem/drug effects , Meristem/metabolism , Methyl Methanesulfonate/antagonists & inhibitors , Mitotic Index , Onions/cytology , Onions/metabolismABSTRACT
(R)-Goniothalamin (R-GNT) is a secondary metabolite isolated from the plants of the genus Goniothalamus. This molecule has attracted the attention of researchers because of its selective cytotoxicity against tumor cells and its ability to induce apoptosis. (S)-Goniothalamin (S-GNT) is a synthetic enantiomer of R-GNT, and its mechanism of action is largely unknown. In this study, we investigated the activity of S-GNT in a human non-small cell lung cancer NCI-H460 cells. We observed that the cells exposed to this compound exhibited cytotoxicity in a concentration-dependent manner. Based on the data obtained through the assessment of apoptosis induction in situ and the comet assay, we suggest that this cytotoxicity occurs due to the potential ability of this molecule to induce DNA damage with the consequent induction of cell death via apoptosis. A significant reduction in the messenger RNA levels of baculoviral inhibitor of apoptosis repeat-containing 5 (BIRC5) gene that encodes the survivin protein was found. This novel finding may explain the inhibition of cell proliferation and induction of apoptosis in tumor cells caused by this compound.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Carcinoma, Non-Small-Cell Lung/drug therapy , DNA Damage , Down-Regulation/drug effects , Inhibitor of Apoptosis Proteins/antagonists & inhibitors , Pyrones/pharmacology , Carcinoma, Non-Small-Cell Lung/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Comet Assay , Humans , Inhibitor of Apoptosis Proteins/genetics , Inhibitor of Apoptosis Proteins/metabolism , Kinetics , Lethal Dose 50 , Lung Neoplasms/drug therapy , Lung Neoplasms/metabolism , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , RNA, Messenger/metabolism , Stereoisomerism , SurvivinABSTRACT
Cisplatin is an effective antineoplastic drug. However, it provokes considerable collateral effects, including genotoxic and clastogenic activity. It has been reported that a diet rich in glutamine can help inhibit such collateral effects. We evaluated this activity in 40 Swiss mice, distributed into eight experimental groups: G1 - Control group (PBS 0.1 mL/10 g body weight); G2 - cisplatin group (cisplatin 6 mg/kg intraperitoneally); G3, G4, G5 - glutamine groups (glutamine at 150, 300, and 600 mg/kg, respectively; orally); G6, G7, G8 - Pre-treatment groups (glutamine at 150, 300, and 600 mg/kg, respectively; orally and cisplatin 6 mg/kg intraperitonially). For the micronucleus assay, samples of blood were collected (before the first use of the drugs at T0, then 24 (T1) and 48 (T2) hours after the first administration). For the comet assay, blood samples were collected only at T2. The damage reduction percentages for the micronucleus assay were 90.0, 47.3, and 37.3% at T1 and 46.0, 38.6, and 34.7% at T2, for G6, G7, and G8 groups, respectively. For the comet assay, the damage reduction percentages were 113.0, 117.4, and 115.0% for G6, G7, and G8, respectively. We conclude that glutamine is able to prevent genotoxic and clastogenic damages caused by cisplatin.
Subject(s)
Antimutagenic Agents/pharmacology , Antineoplastic Agents/toxicity , Cisplatin/toxicity , DNA Damage , Glutamine/pharmacology , Animals , Antimutagenic Agents/therapeutic use , Antineoplastic Agents/therapeutic use , Cisplatin/therapeutic use , Glutamine/therapeutic use , Leukocytes, Mononuclear/drug effects , Male , Mice , Micronucleus Tests , Mutagens/therapeutic use , Mutagens/toxicityABSTRACT
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.
Subject(s)
Antimalarials/pharmacology , Artemisinins/pharmacology , Caspase 3/genetics , DNA Damage , Lactones/pharmacology , Superoxide Dismutase/genetics , Antimalarials/adverse effects , Antimalarials/toxicity , Artemisinins/adverse effects , Artemisinins/toxicity , Artesunate , Caspase 3/metabolism , Dose-Response Relationship, Drug , Hep G2 Cells , Humans , Lactones/adverse effects , Lactones/toxicity , Superoxide Dismutase/metabolism , Superoxide Dismutase-1 , Transcription, Genetic/drug effectsABSTRACT
The incidence of colorectal cancer is growing worldwide. The characterization of compounds present in the human diet that can prevent the occurrence of colorectal tumors is vital. The oligosaccharide inulin is such a compound. The aim of this study was to evaluate the antigenotoxic, antimutagenic and anticarcinogenic effects of inulin in vivo. Our study is based on 3 assays that are widely used to evaluate chemoprevention (comet assay, micronucleus assay, and aberrant crypt focus assay) and tests 4 protocols of treatment with inulin (pre-treatment, simultaneous, post-treatment, and pre + continuous). Experiments were carried out in Swiss male mice of reproductive age. In order to induce DNA damage, we used the pro-carcinogenic agent 1,2-dimethylhydrazine. Inulin was administered orally at a concentration of 50 mg/kg body weight following the protocols mentioned above. Inulin was not administered to the control groups. Our data from the micronucleus assay reveal antimutagenic effects of inulin in all protocols. The percentage of inulin-induced damage reduction ranged from 47.25 to 141.75% across protocols. These data suggest that inulin could act through desmutagenic and bio-antimutagenic mechanisms. The anticarcinogenic activity (aberrant crypt focus assay) of inulin was observed in all protocols and the percentages of damage reduction ranged from 55.78 to 87.56% across protocols. Further tests, including human trials, will be necessary before this functional food can be proven to be effective in the prevention and treatment of colon cancer.
Subject(s)
Aberrant Crypt Foci/prevention & control , Antineoplastic Agents/therapeutic use , Inulin/therapeutic use , Administration, Oral , Animals , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/pharmacology , Chemoprevention , Colorectal Neoplasms/prevention & control , DNA Damage/drug effects , Inulin/administration & dosage , Inulin/pharmacology , Male , Mice , Micronuclei, Chromosome-Defective/drug effectsABSTRACT
Roots of Galianthe thalictroides K. Schum. (Rubiaceae) are used in folk medicine in the State of Mato Grosso do Sul, Brazil, for treating and preventing cancer. To gain information about the genotoxicity of extracts (aqueous and EtOH), the CHCl3 phase resulting from partition of the EtOH extract and the indole monoterpene alkaloid 1 obtained from this plant. The genotoxicity of 1 and extracts was evaluated in vivo through the Drosophila melanogaster wing Somatic Mutation and Recombination Test - SMART, while in vitro cytotoxic (MTT) and Comet assays were performed only with alkaloid 1. The results obtained with the SMART test indicated that the aqueous extract had no genotoxic activity. The EtOH extract was not genotoxic to ST descendants but genotoxic to HB ones. The CHCl3 phase was genotoxic and cytotoxic. Alkaloid 1 showed significant mutational events with SMART, in the cytotoxicity assay (MTT), it showed a high cytotoxicity for human hepatoma cells (HepG2), whereas for the Comet assay, not showing genotoxic activity. The ethanol extract was shown to be genotoxic to HB descendants in the SMART assay, while the results obtained in this test for the monoterpene indole alkaloid 1 isolated from this extract.
Subject(s)
Antineoplastic Agents, Phytogenic/adverse effects , Hepatocytes/drug effects , Indole Alkaloids/adverse effects , Monoterpenes/adverse effects , Plant Extracts/adverse effects , Rubiaceae/chemistry , Animals , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Antineoplastic Agents, Phytogenic/pharmacology , Brazil , Carcinoma, Hepatocellular/drug therapy , Cell Survival/drug effects , Comet Assay , Ethnopharmacology , Hep G2 Cells , Humans , Indole Alkaloids/chemistry , Indole Alkaloids/isolation & purification , Indole Alkaloids/pharmacology , Liver Neoplasms/drug therapy , Molecular Structure , Monoterpenes/chemistry , Monoterpenes/isolation & purification , Monoterpenes/pharmacology , Mutagenicity Tests , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Extracts/pharmacology , Plant Roots/chemistry , Solubility , Solvents/chemistry , Temperature , Tubulin Modulators/adverse effects , Tubulin Modulators/chemistry , Tubulin Modulators/isolation & purification , Tubulin Modulators/pharmacologyABSTRACT
Previous studies in rodents treated with the pro-carcinogen 1,2-dimethylhydrazine suggested that the consumption of wheat bran protected against DNA damage in the colon and rectum. Based on this information, we evaluated wheat bran as a functional food in the prevention and treatment of colon cancer. We used the aberrant crypt focus assay to evaluate the anticarcinogenic potential of wheat bran (Triticum aestivum variety CD-104), the comet assay to evaluate its antigenotoxicity potential, and the micronucleus assay to evaluate its antimutagenic potential. The wheat bran gave good antimutagenic and anticarcinogenic responses; the DNA damage decreased from 90.30 to 26.37% and from 63.35 to 28.73%, respectively. However, the wheat bran did not significantly reduce genotoxicity. Further tests will be necessary, including tests in human beings, before this functional food can be recommended as an adjunct in the prevention and treatment of colon cancer.
Subject(s)
Anticarcinogenic Agents/pharmacology , Antimutagenic Agents/pharmacology , Dietary Fiber/pharmacology , Animals , Colon/drug effects , Colon/pathology , DNA Damage , Humans , Male , Mice , Micronucleus Tests , Organ Size/drug effects , Weight Gain/drug effectsABSTRACT
Studies show that soy imparts many favorable properties in the human body, including the prevention of chronic diseases such as osteoporosis, heart disease, cancer, and diabetes. Soy is rich in isoflavones, and it is a candidate for the chemoprevention of diseases owing to its low toxicity. In this study, a soy phytoestrogen (with high levels of the isoflavones genistin and daidzein) was tested in mice to investigate its mutagenicity and genotoxicity using micronucleus and comet assays of mouse peripheral blood. Phytoestrogen (0.083, 0.83 and 8.3 mg/kg body weight) was evaluated with and without the chemotherapeutic agent cyclophosphamide. For the micronucleus assay, blood was collected before treatment and after 24 and 48 h. For the comet assay, blood was collected only after 24 h. Phytoestrogen was not mutagenic and reduced cyclophosphamide-induced DNA damage. The results from the comet assay revealed a reduction of DNA damage; however, phytoestrogen did induce genotoxic damage during the 24-h treatment. This genotoxic damage could have been repaired and was therefore not identified in the micronucleus assay, which detects mutations. The results suggested that the reduction of DNA damage observed in associated treatments could also reduce the side effects of chemotherapy. Moreover, they suggested that phytoestrogen might be a candidate of interest for the chemoprevention of cancer because it protects against DNA damage.
Subject(s)
Comet Assay/methods , Glycine max/chemistry , Micronuclei, Chromosome-Defective/drug effects , Phytoestrogens/pharmacology , Animals , Antimutagenic Agents/pharmacology , Antineoplastic Agents, Alkylating/pharmacology , Cyclophosphamide/pharmacology , DNA/blood , DNA/genetics , DNA Damage/drug effects , Dose-Response Relationship, Drug , Drug Evaluation, Preclinical , Male , Mice , Micronucleus Tests/methods , Mutagens/pharmacologyABSTRACT
The polysaccharide ß-glucan has biological properties that stimulate the immune system and can prevent chronic pathologies, including cancer. It has been shown to prevent damage to DNA caused by the chemical and physical agents to which humans are exposed. However, the mechanism of ß-glucan remains poorly understood. The objective of the present study was to verify the protective effect of ß-glucan on the expression of the genes ERCC5 (involved in excision repair of DNA damage), CASP9 (involved in apoptosis), and CYP1A1 (involved in the metabolism of xenobiotics) using real-time polymerase chain reaction and perform metabolic profile measurements on the HepG2 cells. Cells were exposed to only benzo[a]pyrene (B[a]P), ß-glucan, or a combination of B[a]P with ß-glucan. The results demonstrated that 50 µg/mL ß-glucan significantly repressed the expression of the ERCC5 gene when compared with the untreated control cells in these conditions. No change was found in the CASP9 transcript level. However, the CYP1A1 gene expression was also induced by HepG2 cells exposed to B[a]P only or in association with ß-glucan, showing its effective protector against damage caused by B[a]P, while HepG2 cells exposed to only ß-glucan did not show CYP1A1 modulation. The metabolic profiles showed moderate bioenergetic metabolism with an increase in the metabolites involved in bioenergetic metabolism (alanine, glutamate, creatine and phosphocholine) in cells treated with ß-glucan and to a lesser extent treated with B[a]P. Thus, these results demonstrate that the chemopreventive activity of ß-glucan may modulate bioenergetic metabolism and gene expression.
Subject(s)
Agaricus/chemistry , Caspase 9/metabolism , Cytochrome P-450 CYP1A1/metabolism , DNA-Binding Proteins/metabolism , Endonucleases/metabolism , Nuclear Proteins/metabolism , Transcription Factors/metabolism , beta-Glucans/chemistry , beta-Glucans/pharmacology , Benzo(a)pyrene/toxicity , Caspase 9/genetics , Cytochrome P-450 CYP1A1/genetics , DNA Damage , DNA-Binding Proteins/genetics , Endonucleases/genetics , Energy Metabolism , Gene Expression Regulation/drug effects , Hep G2 Cells , Humans , Nuclear Proteins/genetics , Transcription Factors/genetics , TranscriptomeABSTRACT
Phenols are a large and diverse class of compounds, many of which occur naturally in a variety of food plants; they exhibit a wide range of biological effects, including antibacterial, anti-inflammatory, antiallergic, hepatoprotective, antithrombotic, antiviral, anticarcinogenic, and vasodilatory actions. We examined the genotoxic and clastogenic potential of three phenolic compounds: caffeic, cinnamic and ferulic acids, using the comet and micronucleus assays in vitro. Drug-metabolizing rat hepatoma tissue cells (HTCs) were used. Three different concentrations (50, 500 and 1500 µM) of these phenolic acids were tested on the HTCs for 24 h. The caffeic, cinnamic and ferulic acids were not genotoxic by the comet assay (P > 0.05). However, the micronucleus test showed an increase in the frequency of micronucleated cells for the three compounds, indicating that these substances have clastogenic effects in HTC.
Subject(s)
Caffeic Acids/toxicity , Cinnamates/toxicity , Coumaric Acids/toxicity , Mutagens/toxicity , Animals , Comet Assay , In Vitro Techniques , Rats , Tumor Cells, CulturedABSTRACT
Paracoccidioidomycosis (PCM) is a systemic mycosis caused by the fungus Paracoccidioides brasiliensis (Pb). The cyclosporin A (CsA) is an immunosuppressant drug that inhibits calcineurin and has been described as a potential antifungal drug. The present study investigated the effect of CsA on the immune response, fungal load/antigenemia in experimental murine PCM. It was used four groups of BALB/c mice: (a) infected with 1 x 105 Pb18 yeast cells (Pb), (b) infected and treated with CsA every other day 10 mg/kg of CsA (s.c.) during 30 days (Pb/CsA), (c) treated with CsA (CsA) and (d) no infected/treated (PBS). The immune response was evaluated by lymphocyte proliferation, DTH assays to exoAgs, ELISA for IgG anti-gp43 (specific immune responses) and cytokine serum levels (IFN-γ, TNF-α, IL-4 and IL-10). Fungal load was determined by lung colony-forming units (CFU) counts, lung and liver histopathology analysis and antigenemia determined by inhibition-ELISA. As expected, CsA was able to inhibit the specific cellular and humoral immune response (P < 0.05), with decrease in serum IFN-γ, TNF-α and IL-4 levels (P < 0.05). Cyclosporin A treatment also resulted in significantly decreased lung Pb CFU (P < 0.05) as well as a lower number of yeasts in the lung and liver (P < 0.05) by histopathology. In concordance, the decreased antigenemia was observed in Pb/CsA group (P < 0.05). In conclusion, even with immunosuppressive action, treatment with CsA results in decreased lung fungal load/antigenemia in experimental PCM in BALB/c mice. Further study is required to determine whether this represents less severe disease or protection by CsA.
Subject(s)
Cyclosporine/therapeutic use , Lung/microbiology , Paracoccidioides , Paracoccidioidomycosis/drug therapy , Animals , Antibodies, Fungal/blood , Antigens, Fungal/blood , Antigens, Fungal/immunology , Colony Count, Microbial , Cyclosporine/immunology , Enzyme-Linked Immunosorbent Assay , Fungal Proteins/blood , Fungal Proteins/immunology , Glycoproteins/blood , Glycoproteins/immunology , Hypersensitivity, Delayed/immunology , Immunoglobulin G/blood , Interferon-gamma/blood , Interleukin-10/blood , Interleukin-4/blood , Liver/microbiology , Liver/pathology , Lung/pathology , Lymphocyte Activation , Lymphocytes/immunology , Male , Mice , Mice, Inbred BALB C , Paracoccidioides/drug effects , Paracoccidioides/growth & development , Paracoccidioides/immunology , Paracoccidioidomycosis/immunology , Paracoccidioidomycosis/microbiology , Paracoccidioidomycosis/pathology , Tumor Necrosis Factor-alpha/bloodABSTRACT
The treatment of textile effluents by the conventional method based on activated sludge followed by a chlorination step is not usually an effective method to remove azo dyes, and can generate products more mutagenic than the untreated dyes. The present work evaluated the efficiency of conventional chlorination to remove the genotoxicity/mutagenicity of the azo dyes Disperse Red 1, Disperse Orange 1, and Disperse Red 13 from aqueous solutions. The comet and micronucleus assays with HepG2 cells and the Salmonella mutagenicity assay were used. The degradation of the dye molecules after the same treatment was also evaluated, using ultraviolet and visible absorption spectrum measurements (UV-vis), high performance liquid chromatography coupled to a diode-array detector (HPLC-DAD), and total organic carbon removal (TOC) analysis. The comet assay showed that the three dyes studied induced damage in the DNA of the HepG2 cells in a dose-dependent manner. After chlorination, these dyes remained genotoxic, although with a lower damage index (DI). The micronucleus test showed that the mutagenic activity of the dyes investigated was completely removed by chlorination, under the conditions tested. The Salmonella assay showed that chlorination reduced the mutagenicity of all three dyes in strain YG1041, but increased the mutagenicity of Disperse Red 1 and Disperse Orange 1 in strain TA98. With respect to chemical analysis, all the solutions showed rapid discoloration and a reduction in the absorbance bands characteristic of the chromophore group of each dye. However, the TOC was not completely removed, showing that chlorination of these dyes is not efficient in mineralizing them. It was concluded that conventional chlorination should be used with caution for the treatment of aqueous samples contaminated with azo dyes.