phytoestrogens and thiazolyl-blue

Phytoestrogens are naturally occurring, plant-derived, nonsteroidal phytochemicals with anticarcinogenic potential. The aim of this study was to isolate phytoestrogens from the flax root of Linum usitatissimum and to test their effect on cellular metabolism in the human mammalian carcinoma cell line MCF-7 using the Bionas 2500 analysis system.. Metabolically relevant parameters such as acidification, oxygen consumption and cell adhesion were registered continuously over 8 and 24 hours on six sensor chips in parallel at different concentrations of flax root extracts.. The extracts from flax roots of L. usitatissimum reduced extracellular acidification, respiration and adhesion in a concentration-dependent manner.. The Bionas 2500 analysis system allows multiparametric online monitoring of cellular processes and can be used to detect the mode of action of anticarcinogenic compounds in cellular metabolism.

Topics: Biosensing Techniques; Cell Adhesion; Cell Line, Tumor; Computational Biology; Equipment Design; Flax; Gene Expression Regulation; Humans; Metabolism; Oligonucleotide Array Sequence Analysis; Oxygen Consumption; Phytoestrogens; Plant Extracts; Tetrazolium Salts; Thiazoles; Time Factors

Although the effects of coumestrol on osteoblasts and osteoclasts can be summarized as increasing the bone density and preventing bone resorption, direct and detailed effects of coumestrol on bone marrow stromal cells remain obscure. In the present study, the effects of coumestrol on proliferation and osteoblastic differentiation of rat bone marrow stromal cells (BMSCs) have been investigated; the regulative effect of coumestrol on BMSCs and skeletal system has also been discussed. The results showed that treatment with coumestrol increased cellular activities (analyzed by MTT assay), alkaline phosphatase (ALP), type I collagen and osteocalcin (OCN) activity as well as the protein and gene expression of OPG, gene expression ratio of OPG/RANKL and gene expression of estrogen receptor alpha(ERalpha). These results demonstrate that phytoestrogen coumestrol has a direct enhancing effect on the proliferation and osteogenic differentiation of bone marrow stromal cells, which would lead to stimulation of bone formation, and it can also protect the whole skeletal system by regulating OPG/RANKL expression, and these effects may be mediated by ERalpha.

Topics: Animals; Bone Marrow Cells; Cell Differentiation; Cell Proliferation; Coumestrol; Gene Expression Regulation; Osteoblasts; Osteogenesis; Osteoprotegerin; Phytoestrogens; RANK Ligand; Rats; Receptors, Estrogen; Stromal Cells; Tetrazolium Salts; Thiazoles

This study is to examine the effects of equol on the H(2)O(2)-induced death of bovine aortic endothelial cells (bAECs) and the mechanism of its protective effects. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] assay showed that in the control group, cell survival rate decreased significantly, each in proportion to the duration of the H(2)O(2) stimulation (P<0.05), but, in the equol-pretreated group, such decrease was not statistically significant. After Hoechst 33342 staining, in the equol-pretreated group the number of cells with apoptotic morphology decreased significantly. Equol pretreatment effectively inhibited the H(2)O(2)-induced cell death by the reduction of intracellular ROS production (P<0.05). Incubation of bAECs with equol increased the expression of phospho-p38 MAPK and Bcl-2 after the H(2)O(2) exposure compared with their expression without the equol pretreatment. Furthermore, SB203580 inhibited phospho-p38 MAPK expression and increased apoptotic cell death. This study proves equol has a significant antioxidant effect on the bAECs that were exposed to H(2)O(2).

Topics: Animals; Antioxidants; Aorta; Apoptosis; Cattle; Cytoprotection; Endothelium, Vascular; Equol; Hydrogen Peroxide; Imidazoles; Isoflavones; p38 Mitogen-Activated Protein Kinases; Phytoestrogens; Proto-Oncogene Proteins c-bcl-2; Pyridines; Reactive Oxygen Species; Tetrazolium Salts; Thiazoles

Anti-estrogenic activity of (-)-sesamin (1), helioxanthin (2), savinin (3), taiwanin C (4), and 3-(3,4-dimethoxybenzyl)-2-(3,4-methylenedioxybenzyl)butyrolactone (5) isolated from the root of Acanthopanax chiisanensis was tested using Ishikawa cells. Among them, compound 3 exhibited anti-estrogenic activity (IC50 = 4.86 microM).

Topics: Adenosine Triphosphatases; Antineoplastic Agents, Phytogenic; Cell Line, Tumor; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Eleutherococcus; Enzyme Induction; Estrogen Antagonists; Female; Humans; Lignans; Magnetic Resonance Spectroscopy; Phytoestrogens; Plant Roots; Spectrometry, Mass, Electrospray Ionization; Tetrazolium Salts; Thiazoles

In search of novel therapeutic approaches for Alzheimer's disease (AD), we report herein the identification, design, synthesis, and pharmacological activity of (4-ethyl-piperaz-1-yl)-phenylmethanone derivatives with neuroprotective properties against beta-amyloid-induced toxicity. (4-ethyl-piperaz-1-yl)-phenylmethanone is a common substructure shared by molecules isolated from plants of the Asteraceae genus, traditionally used as restorative of lost or declining mental functions. (4-Ethyl-piperaz-1-yl)-phenylmethanone displayed strong neuroprotective properties against Abeta1-42 and reversed Abeta1-42-induced ATP depletion on neuronal cells, suggesting a mitochondrial site of action. Abeta1-42 has been described to induce a hyperactivity of the glutamate network in neuronal cells. (4-Ethyl-piperaz-1-yl)-phenylmethanone also inhibited the neurotoxic effect that glutamate displayed on PC12 cells, suggesting that the reduction of glutamate-induced neurotoxicity may be one of the mechanisms by which this compound exerts its neuroprotective properties against the deleterious effects of the Abeta1-42. These data suggest that the identified (4-ethyl-piperaz-1-yl)-phenylmethanone chemical entity exerts neuroprotective properties and may serve as a lead compound for the development of novel therapies for AD.

Topics: Adenosine Triphosphate; Amyloid beta-Peptides; Analysis of Variance; Animals; Cell Survival; Chromatography, Thin Layer; Dizocilpine Maleate; Dose-Response Relationship, Drug; Drug Interactions; Free Radicals; Neuroprotective Agents; PC12 Cells; Peptide Fragments; Phytoestrogens; Piperazines; Plant Preparations; Rats; Tetrazolium Salts; Thiazoles

The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay is a widely used screening method to measure cell viability and proliferation. When testing the effects of kaempferol on breast cancer cell number (crystal violet staining) and viability (MTT tetrazolium assay) conflicting results were obtained. Cell number decreased but MTT formazan formation increased, suggesting a direct interaction of kaempferol with the MTT tetrazolium reduction. Direct reductive potential was observed in a cell-free system for the presumptive phytoestrogens kaempferol and resveratrol, and extracts of Hypericum perforatum L. and Cimicifuga racemosa L. All agents led to instantaneous dark blue formazan formation in the absence of cells. Additionally, antioxidants such as ascorbic acid, vitamin E and N-acetylcysteine interfered with the MTT tetrazolium assay. When MCF7 and HS578 cells treated with kaempferol were washed before addition of MTT tetrazolium, the direct reduction of dye was reduced significantly. These results indicate that the MTT tetrazolium assay may lead to false positive results when testing natural compounds with intrinsic reductive potential.

Topics: Acetylcysteine; Antioxidants; Ascorbic Acid; Breast Neoplasms; Cell Division; Cell Survival; Drug Interactions; Estrogens, Non-Steroidal; Flavonoids; Humans; Isoflavones; Kaempferols; Phytoestrogens; Plant Extracts; Plant Preparations; Plants; Quercetin; Resveratrol; Stilbenes; Tetrazolium Salts; Thiazoles; Tumor Cells, Cultured; Vitamin E