ABSTRACT
Ginkgo biloba is a dioecious tree that has been used in traditional Chinese medicine for about 5,000 years. In previous studies on Ginkgo biloba extract (EGb761) using in vitro systems, we confirmed that EGb761 has biphasic effects on estrogenicity. In this study, we evaluated the agonistic and antagonistic activities of EGb761 using a uterotrophic assay in immature female rats. To evaluate agonistic and antagonistic effects of EGb761 on uterus, 21-day-old immature Sprague-Dawley (SD) female rats were treated with EGb761 (100, 200, or 400 mg/kg) by oral gavage, 10 μg/kg of estradiol (E2) or 1 mg/kg tamoxifen (TM) by subcutaneous injection, or with EGb761 plus E2 or TM for 3 consecutive days. At the end of the treatment period, animals were sacrificed and their body weights and organ weights (liver, lung, spleen and kidney) were measured. In addition, estrogen-related gene expressions (IGFBP-1 in liver and CaBP-9 in uterus) were determined. During the experiment, no animal showed clinical signs, a change in body weight or died. EGb761 treatment alone had no effect on absolute/relative uterine weight, luminal epithelial cell height (LECH, μm), or luminal circumference (LC, μm). In addition, uterine weights, LECHs, and LC induced by E2 or TM were not significantly changed by EGb761 at any dose. These results collectively suggested EGb761 has no agonistic/antagonistic effects in utero.
Subject(s)
Animals , Female , Humans , Rats , Body Weight , Epithelial Cells , Estradiol , Estrogens , Gene Expression , Ginkgo biloba , In Vitro Techniques , Injections, Subcutaneous , Liver , Lung , Medicine, Chinese Traditional , Organ Size , Phenobarbital , Rats, Sprague-Dawley , Spleen , Tamoxifen , Trees , Uterus , Weights and MeasuresABSTRACT
Ginkgo biloba is a dioecious tree that has been used in traditional Chinese medicine for about 5,000 years. In previous studies on Ginkgo biloba extract (EGb761) using in vitro systems, we confirmed that EGb761 has biphasic effects on estrogenicity. In this study, we evaluated the agonistic and antagonistic activities of EGb761 using a uterotrophic assay in immature female rats. To evaluate agonistic and antagonistic effects of EGb761 on uterus, 21-day-old immature Sprague-Dawley (SD) female rats were treated with EGb761 (100, 200, or 400 mg/kg) by oral gavage, 10 μg/kg of estradiol (E2) or 1 mg/kg tamoxifen (TM) by subcutaneous injection, or with EGb761 plus E2 or TM for 3 consecutive days. At the end of the treatment period, animals were sacrificed and their body weights and organ weights (liver, lung, spleen and kidney) were measured. In addition, estrogen-related gene expressions (IGFBP-1 in liver and CaBP-9 in uterus) were determined. During the experiment, no animal showed clinical signs, a change in body weight or died. EGb761 treatment alone had no effect on absolute/relative uterine weight, luminal epithelial cell height (LECH, μm), or luminal circumference (LC, μm). In addition, uterine weights, LECHs, and LC induced by E2 or TM were not significantly changed by EGb761 at any dose. These results collectively suggested EGb761 has no agonistic/antagonistic effects in utero.
Subject(s)
Animals , Female , Humans , Rats , Body Weight , Epithelial Cells , Estradiol , Estrogens , Gene Expression , Ginkgo biloba , In Vitro Techniques , Injections, Subcutaneous , Liver , Lung , Medicine, Chinese Traditional , Organ Size , Phenobarbital , Rats, Sprague-Dawley , Spleen , Tamoxifen , Trees , Uterus , Weights and MeasuresABSTRACT
Zinc pyrithione (ZP) is commonly used to prevent dandruff and seborrheic dermatitis. Many consumers are exposed daily to high doses of ZP, causing serious concerns about its toxicity. The reproductive and developmental toxicities were previously reported in pregnant rats. However, the estrogenic activity of ZP at varying degrees of exposure has been rarely studied. Thus, we performed an uterotrophic assay, E-screen assay, and gene expression profiling to assess the estrogenic activity of ZP. For the uterotrophic assay, ZP (2, 10, or 50 mg/kg/d) was subcutaneously administered to ovariectomized rats every day for three days. Uteri were extracted 24 hours after the last dose. Then, wet and blotted uterine weights were measured. For the E-screen essay, MCF-7 cells (a breast cancer cell line) were exposed to 10⁻⁹ to 10⁻⁶ M of ZP, and cell proliferation was then measured. For the gene expression analysis, changes of gene expression levels in uterine samples taken for the uterotrophic assay were analyzed. In the uterotrophic assay, the concentration of ZP had no significant effect on uterine weight. In the E-screen assay, ZP at any concentration showed no significant increase in MCF-7 cell proliferation, compared to the control group. However, 10⁻⁶ M of ZP significantly reduced cell viability. The changes in gene expression slightly differed between the ZP and control groups. The in vivo and in vitro assays, together with gene expression analysis, demonstrated that ZP showed no significant estrogenic activity.
Subject(s)
Animals , Rats , Breast Neoplasms , Cell Proliferation , Cell Survival , Dandruff , Dermatitis, Seborrheic , Estrogens , Gene Expression , Gene Expression Profiling , In Vitro Techniques , MCF-7 Cells , Uterus , Weights and Measures , ZincABSTRACT
Zinc pyrithione (ZP) is commonly used to prevent dandruff and seborrheic dermatitis. Many consumers are exposed daily to high doses of ZP, causing serious concerns about its toxicity. The reproductive and developmental toxicities were previously reported in pregnant rats. However, the estrogenic activity of ZP at varying degrees of exposure has been rarely studied. Thus, we performed an uterotrophic assay, E-screen assay, and gene expression profiling to assess the estrogenic activity of ZP. For the uterotrophic assay, ZP (2, 10, or 50 mg/kg/d) was subcutaneously administered to ovariectomized rats every day for three days. Uteri were extracted 24 hours after the last dose. Then, wet and blotted uterine weights were measured. For the E-screen essay, MCF-7 cells (a breast cancer cell line) were exposed to 10⁻⁹ to 10⁻⁶ M of ZP, and cell proliferation was then measured. For the gene expression analysis, changes of gene expression levels in uterine samples taken for the uterotrophic assay were analyzed. In the uterotrophic assay, the concentration of ZP had no significant effect on uterine weight. In the E-screen assay, ZP at any concentration showed no significant increase in MCF-7 cell proliferation, compared to the control group. However, 10⁻⁶ M of ZP significantly reduced cell viability. The changes in gene expression slightly differed between the ZP and control groups. The in vivo and in vitro assays, together with gene expression analysis, demonstrated that ZP showed no significant estrogenic activity.
Subject(s)
Animals , Rats , Breast Neoplasms , Cell Proliferation , Cell Survival , Dandruff , Dermatitis, Seborrheic , Estrogens , Gene Expression , Gene Expression Profiling , In Vitro Techniques , MCF-7 Cells , Uterus , Weights and Measures , ZincABSTRACT
Plants of the genus Taraxacum, commonly known as dandelions, are used to treat breast cancer in traditional folk medicine. However, their use has mainly been based on empirical findings without sufficient scientific evidence. Therefore, we hypothesized that dandelions would behave as a Selective estrogen receptor modulator (SERM) and be effective as hormone replacement therapy (HRT) in the postmenopausal women. In the present study, in vitro assay systems, including cell proliferation assay, reporter gene assay, and RT-PCR to evaluate the mRNA expression of estrogen-related genes (pS2 and progesterone receptor, PR), were performed in human breast cancer cells. Dandelion ethanol extract (DEE) significantly increased cell proliferation and estrogen response element (ERE)-driven luciferase activity. DEE significantly induced the expression of estrogen related genes such as pS2 and PR, which was inhibited by tamoxifen at 1 μmol·L(-1). These results indicated that DEE could induce estrogenic activities mediated by a classical estrogen receptor pathway. In addition, immature rat uterotrophic assay was carried out to identify estrogenic activity of DEE in vivo. The lowest concentration of DEE slightly increased the uterine wet weight, but there was no significant effect with the highest concentration of DEE. The results demonstrate the potential estrogenic activities of DEE, providing scientific evidence supporting their use in traditional medicine.
Subject(s)
Animals , Female , Humans , Rats , Breast Neoplasms , Drug Therapy , Metabolism , Cell Proliferation , Estrogen Replacement Therapy , Methods , Gene Expression , MCF-7 Cells , Phytoestrogens , Metabolism , Phytotherapy , Plant Extracts , Pharmacology , Therapeutic Uses , Plant Leaves , Receptors, Estrogen , Metabolism , Selective Estrogen Receptor Modulators , Pharmacology , Taraxacum , UterusABSTRACT
Background & objectives: The mature fruits of Solanum nigrum contains steroidal glycosides. These are often used as vegetable and there are evidences on tribal use of these fruits as an oral contraceptive. The present study was carried out to evaluate the estrogenic potential of S. nigrum fruits by in vitro and in vivo assays. Methods: Defatted methanol extract of dried S. nigrum fruits was column fractionated and the glycoside positive fractions pooled. Definite concentrations of the fraction were used for in vitro and in vivo assays. The effect on cell viability was analyzed in MCF-7 cell lines by MTT assay followed by in vitro evaluation of estrogenicity by hydroxy apatite (HAP) binding assay. The results were further evaluated in vivo by performing uterotrophic assay in ovariectomized mouse models. Results: At low concentration (40 μg/ml), SNGF induced a dose-dependent increase in MCF-7 cell proliferation, while higher extract concentrations (80-320 μg/ml) caused progressive cell growth inhibition. The competitive binding assay using 3H-E2 suggests that this effect is mediated by estrogen receptor. Mouse uterotrophic assay revealed a classical uterotrophic response in ovariectomized mice in response to S. nigrum glycoside fraction (SNGF). SNGF at a dose of 100 mg/kg of body wt induced the maximum height of luminal epithelial cells which indicated an increase of 30.8 per cent over control (P<0.01) with a correlated increase in uterine wet wt (150% increase over control). Higher doses (250 and 500 mg/kg body wt) of SNGF did not induce any uterotrophic effect. Interpretation & conclusions: Our preliminary data demonstrate the hormone like activity of Solanum glycosides both in vitro and in vivo in mouse, which needs to be further explored to evaluate the possible mechanism and clinical implications.
Subject(s)
Analysis of Variance , Animals , Cell Survival/drug effects , Chemical Fractionation , Dose-Response Relationship, Drug , Durapatite/metabolism , Estrogens/pharmacology , Female , Fruit/chemistry , Glycosides/pharmacology , Histological Techniques , India , Methanol , Mice , Microscopy, Fluorescence , Plant Extracts/pharmacology , Solanum nigrum/chemistry , Tetrazolium Salts , Thiazoles , TritiumABSTRACT
Gunnera perpensa (Gunneraceae) is an African plant widely used in traditional medicine. This species is known for its activity involving the female reproductive system, such as inducing or increasing labor, treating female infertility, expelling the placenta and/or preventing post-partum hemorrhage. These properties are probably due to (z)-venusol, a majoritary compound, and its action in conjunction with substances in the whole extract and other natural products. In southern Brazil, a native species Gunnera manicata L. that also belongs to Gunneraceae can be found. In spite of the traditional use of G. perpensa, there is no pharmacological and phytochemical information regarding the South American Gunnera species. Therefore, the aim of this study was to investigate the activity of Brazilian G. manicata aqueous extracts on the reproductive system of immature female Wistar rats through a uterotrophic assay and to verify the presence of (z)-venusol by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Data were analyzed by analysis of variance (ANOVA) and Bonferroni´s post-hoc test (p< 0.01). Results obtained shown that G. manicata extracts did not present in vivo anti or estrogenic activity. Furthermore, (z)-venusol compound was not found. This study represents the first preliminary screening done on the South American G. manicata species.
Gunnera perpensa (Gunneraceae) é uma planta de origem africana extensamente utilizada na medicina tradicional do país. Esta espécie é conhecida por suas atividades no sistema reprodutor feminino, como indução ou aumento do trabalho de parto, tratamento da infertilidade em mulheres, expulsão da placenta e/ou impedimento de hemorragia pós-parto. Tais atividades devem-se, provavelmente, ao sinergismo existente entre o (z)-venusol, composto majoritário, e outros compostos presentes na planta. No sul do Brasil, encontra-se uma espécie nativa, Gunnera manicata L., pertencente à família Gunneraceae. Apesar do uso tradicional de G. perpensa, não há informações farmacológicas e fitoquímicas a respeito da espécie sul Americana de Gunnera. Assim, o objetivo deste estudo foi investigar a atividade de extratos aquosos da espécie brasileira G. manicata no sistema reprodutor de ratas Wistar imaturas através de ensaio uterotrófico e verificar a presença do composto (z)-venusol utilizando-se cromatografia líquida acoplada a espectrômetro de massas em tandem (CL-EM/EM). Para a análise estatística, utilizou-se ANOVA/Bonferroni (p<0,01). Os resultados obtidos demonstraram que os extratos de G. manicata testados não apresentaram atividade anti ou estrogênica in vivo. Na análise química não foi verificada a presença do composto (z)-venusol. Este estudo representa o primeiro screening realizado com a espécie sul-americana G. manicata.
Subject(s)
Animals , Female , Rats , Estrogen Antagonists/agonists , Estrogen Antagonists/analysis , /statistics & numerical data , Estrogens/agonists , Estrogens/analysis , Plants, Medicinal , Plant Roots/chemistry , Analysis of Variance , Clinical Trial , Ethnopharmacology , PharmacognosyABSTRACT
Objective To explore the antagonistic effect of Ziyin Xiehuo Recipe(ZYXHR,Recipe for nourishing kidney yin and purging ministerial fire) on estrogen-like activity of environmental endocrine disruptors,nonylphenol(NP) and mixture of NP and bisphenol A(BPA). Methods Totally 30 three-week-old female SD rats were randomly divided into control group(fed with corn oil),exposed group A (feeding NP100 mg / kg ),exposed group B (feeding NP 50 mg/kg+ BPA 200 mg/kg ),treatment group A (NP 100 mg/kg + ZYXHR),treatment group B (NP 50 mg/kg+ BPA 200 mg/kg + ZYXHR). After 15 days of treatment,the uterine weight gain and PCNA protein expression were detected. Results As compared with those of the control group,Uterine wet weight,uterus organ coefficient,thickness of endometrium and smooth muscle,and the height of endometrial glands and glandular epithelium significantly increased in exposed group A and B (P