ABSTRACT
Plant-derived products can assist in the healing process of dermal wounds. It has been demonstrated that Hancornia speciosa latex present angiogenic, osteogenic, anti-inflammatory, and antioxidant activities. Then, it could contribute to the wound healing process. However, natural products in contact with skin may cause dermatitis. The objective of this work was to evaluate the allergic and irritant potential of H. speciosa serum fraction latex using in vitro assays. The obtained results showed that the H. speciosa serum fraction latex has a slightly irritant potential and is not cytotoxic neither allergenic for human cells. Moreover, we identified a remarkable low amount of proteins in this material in comparison to Hevea brasiliensis latex. This result could explain the non-allergenic potential of H. speciosa serum fraction latex because proteins present in latex are the main responsible for allergy. This biomaterial could be used as a non-allergenic source for development of new medicines.
Produtos derivados de plantas podem auxiliar no processo de cicatrização de feridas cutâneas. Foi demonstrado que o látex de Hancornia speciosa apresenta atividades angiogênicas, osteogênicas, antiinflamatórias e antioxidantes. Então, este biomaterial pode contribuir para o processo de cicatrização de feridas. No entanto, produtos naturais em contato com a pele podem causar dermatites. O objetivo deste trabalho foi avaliar o potencial alérgico e irritante do látex da fração soro de H. speciosa por meio de ensaios in vitro. Os resultados obtidos mostraram que o látex da fração do soro de H. speciosa possui um potencial pouco irritante e não é citotóxico nem alergênico para células humanas. Além disso, foi identificado uma notável baixa quantidade de proteínas neste material em comparação ao látex de Hevea brasiliensis. Esse resultado poderia explicar o potencial não alergênico do látex da fração soro de H. speciosa, pois as proteínas presentes no látex são as principais responsáveis pela alergia. Este biomaterial pode ser utilizado como fonte não alergênica para desenvolvimento de novos medicamentos.
Subject(s)
Apocynaceae/chemistry , Wound Healing/drug effects , In Vitro TechniquesABSTRACT
Abstract Plant-derived products can assist in the healing process of dermal wounds. It has been demonstrated that Hancornia speciosa latex present angiogenic, osteogenic, anti-inflammatory, and antioxidant activities. Then, it could contribute to the wound healing process. However, natural products in contact with skin may cause dermatitis. The objective of this work was to evaluate the allergic and irritant potential of H. speciosa serum fraction latex using in vitro assays. The obtained results showed that the H. speciosa serum fraction latex has a slightly irritant potential and is not cytotoxic neither allergenic for human cells. Moreover, we identified a remarkable low amount of proteins in this material in comparison to Hevea brasiliensis latex. This result could explain the non-allergenic potential of H. speciosa serum fraction latex because proteins present in latex are the main responsible for allergy. This biomaterial could be used as a non-allergenic source for development of new medicines.
Resumo Produtos derivados de plantas podem auxiliar no processo de cicatrização de feridas cutâneas. Foi demonstrado que o látex de Hancornia speciosa apresenta atividades angiogênicas, osteogênicas, antiinflamatórias e antioxidantes. Então, este biomaterial pode contribuir para o processo de cicatrização de feridas. No entanto, produtos naturais em contato com a pele podem causar dermatites. O objetivo deste trabalho foi avaliar o potencial alérgico e irritante do látex da fração soro de H. speciosa por meio de ensaios in vitro. Os resultados obtidos mostraram que o látex da fração do soro de H. speciosa possui um potencial pouco irritante e não é citotóxico nem alergênico para células humanas. Além disso, foi identificado uma notável baixa quantidade de proteínas neste material em comparação ao látex de Hevea brasiliensis. Esse resultado poderia explicar o potencial não alergênico do látex da fração soro de H. speciosa, pois as proteínas presentes no látex são as principais responsáveis pela alergia. Este biomaterial pode ser utilizado como fonte não alergênica para desenvolvimento de novos medicamentos.
ABSTRACT
Abstract Plant-derived products can assist in the healing process of dermal wounds. It has been demonstrated that Hancornia speciosa latex present angiogenic, osteogenic, anti-inflammatory, and antioxidant activities. Then, it could contribute to the wound healing process. However, natural products in contact with skin may cause dermatitis. The objective of this work was to evaluate the allergic and irritant potential of H. speciosa serum fraction latex using in vitro assays. The obtained results showed that the H. speciosa serum fraction latex has a slightly irritant potential and is not cytotoxic neither allergenic for human cells. Moreover, we identified a remarkable low amount of proteins in this material in comparison to Hevea brasiliensis latex. This result could explain the non-allergenic potential of H. speciosa serum fraction latex because proteins present in latex are the main responsible for allergy. This biomaterial could be used as a non-allergenic source for development of new medicines.
Resumo Produtos derivados de plantas podem auxiliar no processo de cicatrização de feridas cutâneas. Foi demonstrado que o látex de Hancornia speciosa apresenta atividades angiogênicas, osteogênicas, antiinflamatórias e antioxidantes. Então, este biomaterial pode contribuir para o processo de cicatrização de feridas. No entanto, produtos naturais em contato com a pele podem causar dermatites. O objetivo deste trabalho foi avaliar o potencial alérgico e irritante do látex da fração soro de H. speciosa por meio de ensaios in vitro. Os resultados obtidos mostraram que o látex da fração do soro de H. speciosa possui um potencial pouco irritante e não é citotóxico nem alergênico para células humanas. Além disso, foi identificado uma notável baixa quantidade de proteínas neste material em comparação ao látex de Hevea brasiliensis. Esse resultado poderia explicar o potencial não alergênico do látex da fração soro de H. speciosa, pois as proteínas presentes no látex são as principais responsáveis pela alergia. Este biomaterial pode ser utilizado como fonte não alergênica para desenvolvimento de novos medicamentos.
Subject(s)
Humans , Apocynaceae , Hevea , Wound Healing , Biocompatible Materials , Allergens , LatexABSTRACT
Plant-derived products can assist in the healing process of dermal wounds. It has been demonstrated that Hancornia speciosa latex present angiogenic, osteogenic, anti-inflammatory, and antioxidant activities. Then, it could contribute to the wound healing process. However, natural products in contact with skin may cause dermatitis. The objective of this work was to evaluate the allergic and irritant potential of H. speciosa serum fraction latex using in vitro assays. The obtained results showed that the H. speciosa serum fraction latex has a slightly irritant potential and is not cytotoxic neither allergenic for human cells. Moreover, we identified a remarkable low amount of proteins in this material in comparison to Hevea brasiliensis latex. This result could explain the non-allergenic potential of H. speciosa serum fraction latex because proteins present in latex are the main responsible for allergy. This biomaterial could be used as a non-allergenic source for development of new medicines.
Subject(s)
Apocynaceae , Hevea , Allergens , Biocompatible Materials , Humans , Latex , Wound HealingABSTRACT
The objective of this study was to evaluate the cytotoxic activity of rosemary (REO, Rosmarinus officinalis L.), turmeric (CEO, Curcuma longa L.), and ginger (GEO, Zingiber officinale R.) essential oils in HeLa cells. Cytotoxicity tests were performed in vitro, using tetrazolium (MTT) and neutral red assays for evaluation of antiproliferative activity by different mechanisms, trypan blue assay to assess cell viability and evaluation of cell morphology for Giemsa to observe the cell damage, and Annexin V to evaluate cell death by apoptosis. CEO and GEO exhibited potent cytotoxic activity against HeLa cells. IC50 obtained was 36.6 µg/mL for CEO and 129.9 µg/mL for GEO. The morphology of HeLa cells showed condensation of chromatin, loss of cell membrane integrity with protrusions (blebs), and cell content leakage for cells treated with CEO and GEO, from the lowest concentrations studied, 32.81 µg/mL of CEO and 32.12 µg/mL of GEO. The Annexin V assay revealed a profile of cell death by apoptosis for both CEO and GEO. The results indicate cytotoxic activity in vitro for CEO and GEO, suggesting potential use as anticancer agents for cervical cancer cells.
Subject(s)
Curcuma/chemistry , Oils, Volatile/pharmacology , Plant Extracts/pharmacology , Rosmarinus/chemistry , Zingiber officinale/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , HeLa Cells , HumansABSTRACT
Several shortcomings of current Parkinson's disease (PD) models limit progress in identification of environmental contributions to disease pathogenesis. The conditionally immortalized cell line LUHMES promises to make human dopaminergic neuronal cultures more easily available, but these cells are difficult to culture for extended periods of time. We overcame this problem by culturing them in 3D with minor medium modifications. The 3D neuronal aggregates allowed penetration by small molecules and sufficient oxygen and nutrient supply for survival of the innermost cells. Using confocal microscopy, gene expression, and flow cytometry, we characterized the 3D model and observed a highly reproducible differentiation process. Visualization and quantification of neurites in aggregates was achieved by adding 2 % red fluorescent protein-transfected LUHMES cells. The mitochondrial toxicants and established experimental PD agents, rotenone and MPP+, perturbed genes involved in one-carbon metabolism and transsulfuration pathways (ASS1, CTH, and SHTM2) as in 2D cultures. We showed, for the first time in LUHMES, down-regulation of mir-7, a miRNA known to target alpha-synuclein and to be involved in PD. This was observed as early as 12 h after rotenone exposure, when pro-apoptotic mir-16 and rotenone-sensitive mir-210 were not yet significantly perturbed. Finally, washout experiments demonstrated that withdrawal of rotenone led to counter-regulation of mir-7 and ASS1, CTH, and SHTM2 genes. This suggests a possible role of these genes in direct cellular response to the toxicant, and the model appears to be suitable to address the processes of resilience and recovery in neurotoxicology and Parkinson's disease in future studies.
Subject(s)
Antiparkinson Agents/pharmacology , Dopaminergic Neurons/drug effects , Drug Evaluation, Preclinical/methods , Neuroprotective Agents/pharmacology , Neurotoxins/toxicity , Toxicity Tests, Acute/methods , Toxicity Tests, Chronic/methods , Cell Aggregation , Cell Culture Techniques , Cell Differentiation , Cell Line, Transformed , Cell Survival/drug effects , Dopaminergic Neurons/cytology , Dopaminergic Neurons/metabolism , Drug Resistance , Gene Expression Regulation/drug effects , Humans , Imaging, Three-Dimensional , Luminescent Proteins/genetics , Luminescent Proteins/metabolism , Microscopy, Confocal , Microscopy, Fluorescence , Nerve Tissue Proteins/agonists , Nerve Tissue Proteins/antagonists & inhibitors , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Neurites/drug effects , Neurites/metabolism , Recombinant Proteins/metabolismABSTRACT
A simple, precise and accurate high-performance liquid chromatographic method has been developed for the determination of lupeol in polymeric nanocapsules. Chromatographic separation was performed on a Varian C8 column (250 mm x 4.6 mm x 5 mm) maintained at 35°C, with a mobile phase composed of acetonitrile and methanol (95:5 v/v) acidified with 0.1% acetic acid, flowing at 1.2 mL/min, with an injected sample volume of 20 µL, UV detection at 210 nm and a run time of 6.2 min. The proposed method was linear over the concentration range 10-250 µg/mL, with R2= 0.9996. Analyses of accuracy and precision showedlow values of relative standard deviation (<4.2%). The methodology was specific, linear, accurate, precise and robust and proved to be adequate for the quantitative analysis of lupeol in polymeric nanocapsules...
Um método de cromatografia líquida de alta performance simples, exato e preciso foi desenvolvido para a determinação do lupeol em nanocápsulas poliméricas. A separação cromatográfica foi realizada numa coluna Varian C8 (250 mm x 4,6 mm x 5 mm), mantida a 35°C, fase móvel constituída por acetonitrila e metanol acidificado com ácido acético a 0,1% (95:5 v/v), e taxa de fluxo de 1,2 mL/min, com um volume injeção de amostra de 20 µl e detecção UV a 210 nm, com o tempo de eluição de 6,2 min. O método proposto é linear para a faixa de concentração de 10 a 250 µg/mL com coeficiente de correlação de 0,9996. As análises de exatidão e precisão demonstraram baixos valores de desvio padrão relativo (< 4,2%). A metodologia foi específica, linear, precisa, exata e robusta, se mostrando capaz de ser aplicada para quantificação de lupeol em nanocápsulas poliméricas...
Subject(s)
Humans , Nanocapsules/analysis , Pentacyclic Triterpenes/isolation & purification , Chromatography, High Pressure LiquidABSTRACT
Duchenne muscular dystrophy (DMD) is still an untreatable lethal X-linked disorder, which affects 1 in 3500 male births. It is caused by the absence of muscle dystrophin due to mutations in the dystrophin gene. The potential regenerative capacity as well as immune privileged properties of mesenchymal Stem Cells (MSC) has been under investigation for many years in an attempt to treat DMD. One of the questions to be addressed is whether stem cells from distinct sources have comparable clinical effects when injected in murine or canine muscular dystrophy animal models. Many studies comparing different stem cells from various sources were reported but these cells were obtained from different donors and thus with different genetic backgrounds. Here we investigated whether human pericytes obtained from 4 different tissues (muscle, adipose tissue, fallopian tube and endometrium) from the same donor have a similar clinical impact when injected in double mutant Utrn (tm1Ked) Dmd (mdx) /J mice, a clinically relevant model for DMD. After a weekly regimen of intraperitoneal injections of 10(6) cells per 8 weeks we evaluated the motor ability as well as the life span of the treated mice as compared to controls. Our experiment showed that only adipose tissue derived pericytes are able to increase significantly (39 days on average) the life span of affected mice. Microarray analysis showed an inhibition of the interferon pathway by adipose derived pericytes. Our results suggest that the clinical benefit associated with intraperitoneal injections of these adult stem cells is related to immune modulation rather than tissue regeneration.
Subject(s)
Adipose Tissue/physiology , Pericytes/physiology , Adipose Tissue/metabolism , Animals , Cell Differentiation/physiology , Cells, Cultured , Disease Models, Animal , Dystrophin/metabolism , Female , Humans , Mesenchymal Stem Cells/metabolism , Mesenchymal Stem Cells/physiology , Mice , Mice, Inbred mdx , Middle Aged , Muscle, Skeletal/metabolism , Muscle, Skeletal/physiology , Muscular Dystrophy, Duchenne/metabolism , Muscular Dystrophy, Duchenne/therapy , Pericytes/metabolismABSTRACT
Duchenne muscular dystrophy (DMD), a lethal X-linked disorder, is the most common and severe form of muscular dystrophies, affecting 1 in 3,500 male births. Mutations in the DMD gene lead to the absence of muscle dystrophin and a progressive degeneration of skeletal muscle. The possibility to treat DMD through cell therapy has been widely investigated. We have previously shown that human adipose-derived stromal cells (hASCs) injected systemically in SJL mice are able to reach and engraft in the host muscle, express human muscle proteins, and ameliorate the functional performance of injected animals without any immunosuppression. However, before starting clinical trials in humans many questions still need to be addressed in preclinical studies, in particular in larger animal models, when available. The best animal model to address these questions is the golden retriever muscular dystrophy (GRMD) dog that reproduces the full spectrum of human DMD. Affected animals carry a mutation that predicts a premature termination codon in exon 8 and a peptide that is 5% the size of normal dystrophin. These dogs present clinical signs within the first weeks and most of them do not survive beyond age two. Here we show the results of local and intravenous injections of hASCs into GRMD dogs, without immunosuppression. We observed that hASCs injected systemically into the dog cephalic vein are able to reach, engraft, and express human dystrophin in the host GRMD dystrophic muscle up to 6 months after transplantation. Most importantly, we demonstrated that injecting a huge quantity of human mesenchymal cells in a large-animal model, without immunosuppression, is a safe procedure, which may have important applications for future therapy in patients with different forms of muscular dystrophies.
Subject(s)
Adipose Tissue/cytology , Dystrophin/metabolism , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Muscular Dystrophy, Duchenne/therapy , Animals , Cells, Cultured , Disease Models, Animal , Dogs , Dystrophin/genetics , Female , Humans , Immunosuppression Therapy , Muscle, Skeletal/metabolism , Muscle, Skeletal/pathologyABSTRACT
Topotecan is an important cytotoxic drug that has gained broad acceptance in clinical use for the treatment of refractory ovarian and small-cell lung cancer. The lactone active form of topotecan can be hydrolyzed in vivo, decreasing the drug's therapeutic efficacy. Lipid encapsulation may promote in vivo stabilization by removing topotecan from aqueous media. Earlier reports of topotecan lipid nanoencapsulation have focused on liposomal encapsulation; however, the higher stability and cost-effectiveness of solid lipid nanoparticles (SLN) highlight the potential of these nanoparticles as an advantageous carrier for topotecan. The initial motivation for this work was to develop, for the first time, solid lipid nanoparticles and nanostructured lipid carriers (NLC) with a high drug loading for topotecan. A microemulsion technique was employed to prepare SLNs and NLCs and produced homogeneous, small size, negatively charged lipid nanoparticles with high entrapment efficiency and satisfactory drug loading. However, low recovery of topotecan was observed when the microemulsion temperature was high and in order to obtain high quality nanoparticles, and precise control of the microemulsion temperature is critical. Nanoencapsulation sustained topotecan release and improved its chemical stability and cytotoxicity. Surprisingly, there were no significant differences between the NLCs and SLNs, and both are potential carriers for topotecan delivery.
Subject(s)
Drug Delivery Systems , Nanoparticles/chemistry , Topoisomerase I Inhibitors/chemistry , Topotecan/chemistry , Cell Survival/physiology , Delayed-Action Preparations , Drug Carriers/chemistry , Drug Compounding , Drug Evaluation, Preclinical , Drug Stability , Emulsions/chemistry , Freeze Drying , Humans , K562 Cells , Lipids/chemistry , Nanoparticles/administration & dosage , Nanoparticles/toxicity , Nanostructures/analysis , Nanostructures/chemistry , Nanostructures/toxicity , Particle Size , Solubility , Surface Properties , Temperature , Topoisomerase I Inhibitors/administration & dosage , Topoisomerase I Inhibitors/metabolism , Topoisomerase I Inhibitors/toxicity , Topotecan/administration & dosage , Topotecan/metabolism , Topotecan/toxicity , Trypan BlueABSTRACT
AIM OF THE STUDY: Palicourea coriacea (Cham.) K Schum, is an endemic plant used in the Midwestern Region of Brazil, popularly known as "douradinha do campo" and "congonha do campo". This plant has been used in traditional medicine for several ailments, especially to treat kidney diseases. Since no formal studies on the biological activities and medicinal properties of the ethanolic extract of Palicourea coriacea (PCEE) have been carried out previously, the present study represents the first research into the efficacy of this plant as a diuretic agent employing laboratory rats as test animals. MATERIALS AND METHODS: For diuretic activity evaluation we assayed three doses of PCEE (20, 40 and 80mg/kg) and measurement of the urinary volume and electrolytes (Na(+), K(+)) concentration were taken. The acute oral toxicity of PCEE was investigated according to OECD Guideline 423. RESULTS: The oral administration of a single dose of PCEE significantly increased the urinary volume in 24h. Additionally, the treatment with PCEE increased, in a dose-dependent manner, the excretion of both, Na(+) and K(+). No sign of toxicity was observed in the animals. CONCLUSIONS: The present study confirmed the ethnopharmacological use of Palicourea coriacea as a diuretic agent in the experimental condition tested here. Additionally, this work supports the importance of the preservation of local knowledge as well as the conservation of Brazilian biodiversity.
Subject(s)
Diuretics/pharmacology , Plant Extracts/pharmacology , Potassium/urine , Rubiaceae , Sodium/urine , Urination/drug effects , Administration, Oral , Animals , Brazil , Diuretics/toxicity , Dose-Response Relationship, Drug , Male , Medicine, Traditional , Plant Extracts/toxicity , Rats , Rats, Wistar , Rubiaceae/toxicityABSTRACT
BACKGROUND: CD8+ and natural killer (NK) cells have been considered the most effective cells in the combat of cancer, contributing to better prognosis and longer survival. METHODS: The aim of this study was to evaluate the population of CD8+ and NK cells, by immunohistochemistry, in samples of oral cavity squamous cell carcinoma (OCSCC) and lip squamous cell carcinoma (LSCC), leukoplakia, actinic cheilitis, and healthy oral mucosa (control). The relationship of CD8+ and NK cells with survival data, lymph node metastasis, tumor size, and proliferative index was also evaluated. RESULTS: The number of peritumoral and intratumoral CD8+ and NK cells was significantly higher in LSCC, when compared with control, pre-malignant lesions, and OCSCC. A higher proportion of peritumoral CD8+ cells demonstrated correlation with a lower neoplastic proliferative index. Moreover, patients with OCSCC with a high density of peritumoral CD8+ cells showed a tendency towards a longer survival time. CONCLUSIONS: The differential CD8+ and NK cells infiltration in oral SCC might reflect a distinctive tumor microenvironment with a favorable local cytotoxic immune response against neoplastic cells.
Subject(s)
CD8-Positive T-Lymphocytes/immunology , Carcinoma, Squamous Cell/immunology , Killer Cells, Natural/immunology , Lip Neoplasms/immunology , Lymphocytes, Tumor-Infiltrating/immunology , Mouth Neoplasms/immunology , CD8-Positive T-Lymphocytes/pathology , Carcinoma, Squamous Cell/pathology , Carcinoma, Squamous Cell/secondary , Cheilitis/immunology , Cheilitis/pathology , Cyclin B1/analysis , Female , Follow-Up Studies , Humans , Immunohistochemistry , Killer Cells, Natural/pathology , Leukoplakia, Oral/immunology , Leukoplakia, Oral/pathology , Lip Neoplasms/pathology , Lymphatic Metastasis/pathology , Lymphocyte Count , Lymphocytes, Tumor-Infiltrating/pathology , Male , Middle Aged , Mitotic Index , Mouth Mucosa/immunology , Mouth Mucosa/pathology , Mouth Neoplasms/pathology , Neoplasm Invasiveness , Neoplasm Staging , Survival RateABSTRACT
The growth kinetics of CdTe colloidal nanocrystals has been analyzed quantitatively by means of dynamic light scattering and photoluminescence measurements. The growth rates, size distributions, critical radii, and diffusion constants have been calculated in the framework of the Sugimoto theoretical model. A two-step diffusion-controlled growth regime has been proposed for the reported synthesis and a set of relations for the time evolution of the size distribution has been derived and discussed in the sense of the size distribution focusing concept.
ABSTRACT
Duchenne muscular dystrophy (DMD) is a human disease characterized by progressive and irreversible skeletal muscle degeneration caused by mutations in genes coding for important muscle proteins. Unfortunately, there is no efficient treatment for this disease; it causes progressive loss of motor and muscular ability until death. The canine model (golden retriever muscular dystrophy) is similar to DMD, showing similar clinical signs. Fifteen dogs were followed from birth and closely observed for clinical signs. Dogs had their disease status confirmed by polymerase chain reaction analysis and genotyping. Clinical observations of musculoskeletal, morphological, gastrointestinal, respiratory, cardiovascular, and renal features allowed us to identify three distinguishable phenotypes in dystrophic dogs: mild (grade I), moderate (grade II) and severe (grade III). These three groups showed no difference in dystrophic alterations of muscle morphology and creatine kinase levels. This information will be useful for therapeutic trials, because DMD also shows significant, inter- and intra-familiar clinical variability. Additionally, being aware of phenotypic differences in this animal model is essential for correct interpretation and understanding of results obtained in pre-clinical trials.
Subject(s)
Muscular Dystrophy, Animal/pathology , Phenotype , Animals , Disease Models, Animal , Dogs , Muscle, Skeletal/pathologySubject(s)
Dystrophin/genetics , Muscular Dystrophy, Animal/genetics , Point Mutation , Alternative Splicing/genetics , Animals , DNA Mutational Analysis , Disease Models, Animal , Dogs , Dystrophin/deficiency , Muscle Strength/genetics , Muscle Strength/physiology , Muscular Dystrophy, Animal/pathology , Muscular Dystrophy, Animal/physiopathologyABSTRACT
Organotellurium(IV) compounds have been reported to have multiple biological activities including cysteine protease-inhibitory activity, mainly cathepsin B. As cathepsin B is a highly predictive indicator for prognosis and diagnosis of cancer, a possible antitumor potential for these new compounds is expected. In this work, it was investigated the effectiveness of organotellurium(IV) RT-04 to produce lethal effects in the human promyelocytic leukaemia cell line HL60. Using the MTT tetrazolium reduction test, and trypan blue exclusion assay, the IC50 for the compound after 24 h incubation was 6.8 and 0.35 microM, respectively. Moreover, the compound was found to trigger apoptosis in HL60 cells, inducing DNA fragmentation and caspase-3, -6, and -9 activations. The apoptsosis-induced by RT-04 is probably related to the diminished Bcl-2 expression, observed by RT-PCR, in HL60-treated cells. In vivo studies demonstrated that the RT-04 treatment (2.76 mg/kg given for three consecutive days) produces no significant toxic effects for bone marrow and spleen CFU-GM. However, higher doses (5.0 and 10 mg/kg) produced a dose-dependent reduction in the number of CFU-GM of RT-04-treated mice. These results suggest that RT-04 is able to induce apoptosis in HL60 cells by Bcl-2 expression down-modulation. Further studies are necessary to better clarify the effects of this compound on bone marrow normal cells.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Gene Expression Regulation, Neoplastic , Genes, bcl-2/drug effects , Organometallic Compounds/pharmacology , Animals , Caspases/metabolism , DNA Fragmentation/drug effects , Dose-Response Relationship, Drug , Down-Regulation , Drug Screening Assays, Antitumor , Enzyme Activation , Gene Expression Regulation, Neoplastic/drug effects , HL-60 Cells , Humans , Inhibitory Concentration 50 , Male , Mice , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
In the present work we evaluated both the mutagenicity and antimutagenicity of the Pothomorphe umbellata root extract (PUE) and its isolated active principle, the 4-nerolidylcatechol (4-NC), in bone marrow cells of mice using the micronucleus test. Swiss male mice were orally treated for 4 days with PUE (200, 100 or 50mg/kg/day) or 4-NC (50, 25 or 12.5mg/kg/day) prior to exposition with a single dose (200mg/kg) of cyclophosphamide (CP), 24h after the end of the treatment. The results demonstrated that the PUE and 4-NC did not have any mutagenic effect on mouse bone marrow cells; quite the opposite, there was a protective effect against genotoxicity induced by cyclophosphamide. Taken together, under the conditions tested herein, mice treated with PUE and 4-NC showed, in a dose-dependent manner, protective effect against CP-induced genotoxicity. Due to their ability to prevent chromosomal damage, with apparent low toxicity and cost, PUE or pure 4-NC are likely to open a field of interest concerning their possible use in clinical applications.
Subject(s)
Antimutagenic Agents , Antineoplastic Agents, Alkylating/antagonists & inhibitors , Antineoplastic Agents, Alkylating/toxicity , Catechols/pharmacology , Cyclophosphamide/antagonists & inhibitors , Cyclophosphamide/toxicity , Mutagens , Piperaceae/chemistry , Animals , Chromosome Aberrations/drug effects , Dose-Response Relationship, Drug , Erythrocytes/drug effects , Erythrocytes/ultrastructure , Male , Mice , Micronucleus Tests , Plant Extracts/chemistry , Plant Roots/chemistryABSTRACT
In previous works, we have demonstrated that the myeloprotective properties of several natural and synthetic compounds are partly responsible for their antitumor activity in the Ehrlich ascites tumor (EAT) model. In this work, we present information that may be useful to the study of pharmacological and toxicological properties of compounds that affect the hematological compartment. Clonogenic studies in EAT-inoculated mice demonstrated a rapid decrease in bone marrow CFU-GM, whereas a progressive increase in splenic CFU-GM and cellularity was observed, followed by splenomegaly. Bone marrow cellularity declined on the third day after tumor challenge, returning to normal values thereafter. Serum from EAT-bearing mice produced detectable colony-stimulating activity in vitro. Similar results were observed with the conditioned medium from Ehrlich tumor cell cultures, but not with the cell-free Ehrlich tumor ascitic fluid. Tumor inoculation also resulted in a more striking depletion in the number of non-adherent cells in long-term bone marrow cell cultures (LTBMCs) with no bone marrow stroma formation. We speculate that the physiological alterations induced by the EAT growth can be used to assess the ability of compounds to modulate the hematopoietic response.
Subject(s)
Antineoplastic Agents/pharmacology , Carcinoma, Ehrlich Tumor/pathology , Drug Design , Hematopoiesis/drug effects , Hematopoietic Stem Cells/drug effects , Immunologic Factors/pharmacology , Animals , Ascitic Fluid/pathology , Bone Marrow Cells/immunology , Bone Marrow Cells/pathology , Carcinoma, Ehrlich Tumor/drug therapy , Carcinoma, Ehrlich Tumor/immunology , Cell Count , Cell Line, Tumor , Colony-Forming Units Assay , Hematopoiesis/immunology , Hematopoietic Stem Cells/immunology , Male , Mice , Mice, Inbred BALB C , Neoplasm Transplantation , Spleen/immunology , Spleen/pathologyABSTRACT
The capacity of hematopoietic tissues to produce and mobilize phagocytes to the site of infection and tumor growth is of central importance to mediate the early immunological response. In this perspective, studies from our laboratory have defined Listeria monocytogenes infection and the Ehrlich ascites tumor (EAT) as useful models to investigate the effects of natural compounds on the growth and differentiation of granulocyte-macrophage progenitor cells (CFU-GM). As expected, a significant reduction in the number of bone marrow CFU-GM was observed in the initial stages of infection with a sublethal dose of Listeria. Similarly, the bone marrow CFU-GM decreased sharply 4 days after the EAT transplantation. Treatment of infected and tumor-bearing mice with 500 and 1,000 mg/kg of Caesalpinia ferrea aqueous extract, given 3 times orally, significantly stimulated myelopoiesis, whereas no effects were observed with the 250 mg/kg dose. Similar results were obtained in normal mice. The administration of the two higher doses of the extract also protected 15-20% of mice from a lethal dose of Listeria and significantly prolonged survival of EAT-bearing mice. In summary, these results demonstrate that C. ferrea extract acts as a positive regulator of myelopoiesis, and suggest that the therapeutic effect of C. ferrea may be partially mediated by this action.
