The attenuating effect of daidzein (DAI) on oxidative toxicity induced by Aroclor 1254 (A1254) was investigated in mouse testicular cells. Cells were exposed to A1254 alone or with DAI. The oxidative damage was estimated by measuring malondialdehyde (MDA) formation, superoxide dismutase (SOD) activity and glutathione (GSH) content. Results show that A1254 induced a decrease of germ cell number, an elevation in thiobarbituric acid reactive substances (TBARS) but a decrease in SOD activity and GSH content. However, simultaneous supplementation with DAI decreased TBARS level and increased SOD activity and GSH content. Consequently, dietary DAI may restore the intracellular antioxidant system to attenuate the oxidative toxicity of A1254 in testicular cells.
Animals ; Chlorodiphenyl (54% Chlorine) ; toxicity ; Hypoxanthine ; toxicity ; Isoflavones ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred ICR ; Oxidation-Reduction ; Testis ; drug effects ; metabolism ; Xanthine Oxidase ; toxicity

Flavonoids, including flavones, flavonols, anthocyanins, flavanones, chalcones, flavan, proanthocyanidins, isoflavonoids and biflavonoids, etc, are natural components in our diet and plants. Several beneficial properties have been attributed to these compounds, including antioxidant, anti-inflammatory and anticarcinogenic effects, etc. Flavonoid preparations are marketed as herbal medicines or dietary supplements for a variety of alleged nontoxic therapeutic effects. However, they have yet to pass controlled clinical trials for efficacy, and their potential for toxicity is an understudied field of research. This review summarizes the current studies on the toxicity induced by flavonoids and gives some advices for ingesting flavonoids.
Animals ; Antioxidants ; toxicity ; Cytochrome P-450 Enzyme System ; metabolism ; Dietary Supplements ; adverse effects ; toxicity ; Drug Interactions ; Flavonoids ; adverse effects ; isolation & purification ; toxicity ; Free Radical Scavengers ; toxicity ; Humans ; Isoflavones ; isolation & purification ; toxicity ; Plants, Medicinal ; chemistry

OBJECTIVETo evaluate the antioxidant activities of different chemical constituents from Astragalus mongholicus Bunge and their protection against xanthine (XA)/xanthine oxidase (XO)-induced toxicity in PC12 cells.

METHODSThe compounds of Astragalus mongholicus Bunge were isolated by chromatography and the structures were elucidated on the basis of spectral data interpretation. Their antioxidant activities were detected by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities in a cell-free system. Meanwhile, the effects against XA/XO-induced toxicity were assessed using MTT assay in PC12 cells.

RESULTSTen principal constituents were isolated and identified as formononetin (I), ononin (II), calycosin (III), calycosin-7-O-beta-D-glucoside (IV), 9,10-dimethoxypterocarpan-3-O-beta-D-glucoside (V), adenosine (VI), pinitol (VII), daucosterol (VIII), beta-sitoster (IX) and saccharose (X) from Astragalus mongholicus Bunge. The compounds I, III, and IV scavenged DPPH free radicals in vitro. Formononetin and calycosin were found to inhibit XA/XO-induced cell injury significantly, with an estimated EC50 of 50 ng/mL.

CONCLUSIONCompound II, VI, and VII are first reported in this plant. Calycosin exhibits the most potent antioxidant activity both in the cell-free system and in the cell system.

Animals ; Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Free Radical Scavengers ; chemistry ; pharmacology ; Free Radicals ; metabolism ; Isoflavones ; chemistry ; pharmacology ; PC12 Cells ; Rats ; Xanthine ; toxicity ; Xanthine Oxidase ; toxicity

OBJECTIVETo study the possible intervention of isoflavones in cytotoxicity induced by cadmium in vascular endothelial cells.

METHODSAn ECV 304 cell line derived from human umbilical vein endothelial cells was adopted. Genistein/daidzein was added prior to or simultaneously with CdCl2, cell viability was determined by MTT assay, and metallothionein mRNA expression was monitored by RT-PCR method.

RESULTSCell viability was higher in isoflavone and CdCl2 co-treated groups than that in CdCl2 treated group, with CdCl2 concentration at 10, 20, 40, and 80 micromol/L, respectively. However this increase was not observed in the group treated with CdCl2 at a concentration of 60 micromol/L. Isoflavones (10(-10) mol/L to 10(-5) mol/L) were added 24 h before cells were challenged with 80 micromol/L CdCl2 for 24 h or simultaneously with 80 micromol/L CdCl2. Genistein increased cell viability only at 10(-5) mol/L, while daidzein caused a dose-dependent increase from 10(-10) mol/L to 10(-5) mol/L in co-treatment with CdCl2. In pre-treatment, genistein (10(-7) to 10(-5) mol/L) increased cell viability whereas only 10(-5) mol/L of daidzein exerted protection. Apparent protection could be found when the cells were pre-treated with 10(-5) mol/L isoflavones for over 12 h, whereas 24 h incubation was required in such a co-treatment, with the exception of daidzein that had a significant protection in only 3 h. Isoflavones (10(-6) mol/L) incubated for 3 h to 24 h, increased MT IIA and MT IF mRNA expression, but the induction could not last for more than 24 h. Co-treatment with isoflavones could induce an additional induction of MT IIA mRNA expression in cells exposed to cadmium. However, the additional induction of MT IIA and MT IF mRNA was not seen when pre-treatment was carried out with isoflavones, with the exception of an increase in MT IIA mRNA expression in the daidzein pre-treated group.

CONCLUSIONGenistein/daidzein could reverse the cytotoxicity of cadmium either in pre-treatment or in co-treatment. The protection is the strongest in 10(-5) mol/L of isoflavones with a dose-dependent pattern. There are differences between genistein and daidzein in their protective effects. Whether the protection of isoflavones is related to their capacity of inducing MT mRNA expression remains to be elucidated.

Cadmium ; toxicity ; Cell Line ; Cell Survival ; drug effects ; Endothelial Cells ; drug effects ; metabolism ; Genistein ; pharmacology ; Humans ; Isoflavones ; pharmacology ; Metallothionein ; genetics ; metabolism ; Protective Agents ; pharmacology ; RNA, Messenger ; metabolism

Inflammation of the prostate can be induced experimentally in rats by the subcutaneous administration of estrogen. However, it is usually achieved at the price of some alteration in the sex steroid hormone balance and morphological changes in the prostate. In this study, a soy-extracted isoflavone mixture with weak estrogenic activity was administered orally in an attempt to induce prostatitis in a more physiologic way and to characterize the inflammation induced. A total of 36 male Sprague-Dawley rats, 8 weeks old, were divided into 2 groups. The control group was fed with only an AIN-76A diet containing no detectable phytoestrogen and the experimental group was fed with AIN-76A and a soy- extracted isoflavone mixture (genistein 60.0% and daidzein 19.6%), 300mg/kg body weight for 9 weeks. The sequential body weight and prostate weight at necropsy were measured. A histologic examination and histomorphometry assessed the changes in the prostate. The serum concentrations of testosterone and dihydrotestosterone were measured to estimate the effects on the androgen level. Intraprostatic concentrations of genistein and daidzein were measured by gas chromatography/ mass spectroscopy (GC/MS). While no sign of prostate inflammation was apparent in the control group, severe inflammatory changes in the stroma, increased epithelial detachment and inflammatory exudates within the glandular lumen of the dorsolateral prostate were observed in more than 80%(15/18) of the experimental group. However, there was no significant difference in the ventral prostate between the two groups. The daidzein and genistein concentrations in both the lateral and ventral prostates were significantly higher in the experimental group than in the control group where no isoflavone was detectable. In addition, the concentrations were much higher in the dorsolateral than in the ventral prostate. Although the body weight gain was not consistent in the experimental group, there were no significant differences in the prostate weight and serum androgen level between groups. In summary, when a soy-extracted genistein and daidzein-rich isoflavone mixture was administered orally into rats, prostatic inflammation with characteristic lobe specificity developed. The present method of inducing prostatitis seems to be a more physiologic than an estrogen-induced experimental model, and sequential pharmacokinetic studies might help in establishing this model as a more valuable tool in assisting future research in this field.
Administration, Oral ; Androgens/blood ; Animal ; Body Weight/drug effects ; Isoflavones/metabolism/*toxicity ; Male ; Organ Weight/drug effects ; Prostatitis/*chemically induced/pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the protective effect of puerarin on the myocardium of rats with acute and chronic alcoholism.

METHODSIn acute alcoholism experiment, normal male SD rats were randomly divided into the control group, alcoholism group and puerarin group (n=8), and high- and low-dose puerarin was administered. In chronic alcoholism experiment, increasing puerarin doses were given. Serum and myocardial levels of spartate aminotransferase (AST) and creatine phosphokinase (CPK) were determined using enzymatic methed, and superoxide dismutase (SOD), malondialdehyde (MDA), Ca(2+)-Mg(2+)-ATPase, and Na(+)-K(+)-ATPase in the myocardium were assayed with colorimetric method. HE staining was used to observe the microscopic changes of the myocardium.

RESULTSCompared with alcoholism group, puerarin-treated groups showed significantly lowered myocardial contents of MDA, CPK and AST and serum levels of AST and CPK (P<0.05, P<0.01) and increased myocardial SOD (P<0.05, P<0.01), Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase activity (P<0.05, P<0.01), but Na(+)-K(+)-ATPase was similar between the two groups (P>0.05). HE staining of the myocardium showed cell swelling and obscure cell boundaries in alcoholism group, especially in chronic alcoholism group. The myocardial structure in puerarin group remained clear and regular.

CONCLUSIONPuerarin can protect from myocardial injuries induced by acute and chronic alcoholism in rats.

Alcoholism ; drug therapy ; Animals ; Cardiomyopathy, Alcoholic ; metabolism ; pathology ; prevention & control ; Ethanol ; toxicity ; Isoflavones ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the protective effects and related mechanisms of soybean isoflavone (SI) on human umbilical vein endothelial cells (HUVECs) injury induced by H₂O₂ and lipopolysaccharide (LPS).

METHODSH₂O₂ and LPS were used to induce HUVECs injury in vitro. Nine experimental groups were examined: control group, H₂O₂ (2 mmol/L for 4 h), LPS (2 mmol/L for 4 h), H₂O₂+low dose SI (1 mg/ml), H₂O₂+moderate dose SI (2.5 mg/ml), H₂O₂+high dose SI (5 mg/ml), LPS+low dose SI (1 mg/ml), LPS+moderate dose SI (2.5 mg/ml), LPS+high dose SI (5 mg/ml). The survival ratio of HUVECs was detected with MTT assay. The cultured cells were loaded by Fura-2/AM and the change of [Ca²⁺] in HUVECs was measured by fluorospectrophotometry. The contents of malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione (GSH-Px) were measured by the commercial kits. The levels of tissue plasminogen activator IL-6 in the supematant were measured by enzyme linked immunosorbent assay (ELISA) kits. Apoptosis rate of the HUVECs was analyzed by flow cytometry.

RESULTSH₂O₂ and LPS significantly decreased HUVECs viability, increased the contents of MDA, IL-6 and decreased the contents of SOD and GSH-Px, and increased the apoptosis rate [(37.8 ± 1.8)% and (38.9 ± 1.1)%]. Co-treatment with SI could reduce MDA and IL-6 while increase SOD and GSH-Px and reduce apoptosis in a dose-dependent manner.

CONCLUSIONThe findings demonstrate that soybean isoflavone could attenuate H₂O₂ and LPS induced injury in human umbilical vein endothelial cells through protecting mitochondrial function, improving antioxygenic activity, and suppressing the mobilization of cytosolic calcium.

Apoptosis ; drug effects ; Calcium ; metabolism ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; pathology ; Humans ; Hydrogen Peroxide ; toxicity ; Isoflavones ; pharmacology ; Lipid Peroxidation ; drug effects ; Lipopolysaccharides ; toxicity ; Oxidative Stress ; drug effects ; Soybeans ; chemistry

OBJECTIVESThis paper aims to investigate the uterotrophic activities of lactational exposure to combination of soy isoflavones (SIF) and bisphenol A (BPA) and to examine estrogen receptor α (ERα) and estrogen receptor β (ERβ) expressions in hypothalamus-pituitary-ovary axis and uterus.

METHODSMaternal rats that were breeding about 8 litters were randomly divided into four groups with seven dams in each group. Dams in different treatment groups received corn oil (control), 150 mg/kg BW of SIF, 150 mg/kg BW of BPA or combination of 150 mg/kg BW of SIF and 150 mg/kg BW of BPA, respectively, from postnatal day 5 to 11 (PND5-11) by gavage. On PND12 and PND70, 10 female litters were killed and hypothalamus, pituitary, ovary and uterus were collected. ERα and ERβ expressions in these organs were detected with Western blotting assay. And vaginal opening time and estrus cycle were examined in animals fed for PND70.

RESULTSOn PND12, the relative uterine weight of rats treated with ISF or BPA or their combination was significantly higher than that of untreated rats (P<0.05). But the relative uterine weight of rats in the co-exposure group was slightly lower than that in the group only exposed to SIF or BPA. On PND 70, however, the relative uterine weight in each treatment group was not statistically different from that in the control group (P>0.05). Vaginal opening time and estrus cycle in groups treated with SIF or BPA or their combination were similar to those in the control group (P>0.05). Exposure to SIF or BPA or their combination could up-regulate or down-regulate ERα and ERβ expressions in hypothalamus, pituitary, ovary and uterus on PND12 and PND70. These regulation patterns for ERα and ERβ were different in different organs at different time points.

CONCLUSIONLactational exposure to ISF or BPA or their combination could induce uterotrophic responses in neonate rats, which disappeared in later life. But these data fail to suggest a possibility for synergic actions between SIF and BPA. It was also demonstrated that the uterotrophic effects of SIF and BPA exposure might, at least, involve modification of ERα or ERβ expressions in the hypothalamus-pituitary-ovary axis.

Animals ; Animals, Newborn ; Benzhydryl Compounds ; Blotting, Western ; Down-Regulation ; Drug Synergism ; Estrogen Receptor alpha ; biosynthesis ; Estrogen Receptor beta ; biosynthesis ; Estrogens, Non-Steroidal ; pharmacokinetics ; toxicity ; Female ; Hypothalamo-Hypophyseal System ; drug effects ; metabolism ; Isoflavones ; isolation & purification ; pharmacokinetics ; toxicity ; Lactation ; metabolism ; Maternal Exposure ; Organ Size ; drug effects ; Ovary ; drug effects ; metabolism ; Phenols ; pharmacokinetics ; toxicity ; Phytoestrogens ; isolation & purification ; pharmacokinetics ; toxicity ; Rats ; Rats, Sprague-Dawley ; Sexual Maturation ; drug effects ; Soybeans ; chemistry ; Up-Regulation ; Uterus ; drug effects ; metabolism

AIMTo study the protective effects of hydroxyethylpuerarin against the injury of astrocytes induced by hydrogen peroxide (H2O2).

METHODSExperiments were performed with cells from passage 4. Plasma membrane integrity was measured by lactate dehydrogenase (LDH) release. The occurrence of apoptosis was measured by flow cytometry. The glutamate uptake of astrocytes was studied with [3H]-glutamate incorporation. Intracellular superoxide dismutase (SOD) activity and malondialdehyde (MDA) level were assessed by automatic biochemistry analyzer.

RESULTSCompared with H2O2 injured group, the occurrence of apoptosis, levels of LDH release and intracellular MDA of astrocytes reduced in hydroxyethylpuerarin pre-treated groups, but the glutamate uptake and intracellular SOD activity of astrocytes increased.

CONCLUSIONHydroxyethylpuerarin could reduce the occurrence of apoptosis and improve neurotrophic function of astrocytes, which may be related with its antioxidant effects during oxidative stress.

Animals ; Animals, Newborn ; Antioxidants ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Astrocytes ; cytology ; drug effects ; metabolism ; Brain ; cytology ; metabolism ; Cells, Cultured ; Glutamic Acid ; metabolism ; Hydrogen Peroxide ; toxicity ; Isoflavones ; isolation & purification ; pharmacology ; L-Lactate Dehydrogenase ; metabolism ; Malondialdehyde ; metabolism ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism

OBJECTIVETo examine whether Chinese medical herb Pueraria crude extract (CP) and standard of pure puerarin (SP) possess the same neuroprotective effects during concomitant ethanol (EtOH) treatment.

METHODSHippocampus cultures were prepared from mice at gestational age of 18 day. Cell viability was measured by MTT assay. RT-PCR was employed to determine mRNA expression of superoxide dismutase (SOD).

RESULTSAs measured by MTT assay, supplementation with 15 mg/L CP or 10 mg/L SP afforded neuroprotection against all EtOH concentrations (50, 200 and 350 mmol/L, respectively) in embryonic hippocampal culture system. In addition, both 15 mg/L CP and 10 mg/L SP could decrease expression of SOD at mRNA level.

CONCLUSIONThis study suggests that CP and SP could decrease oxidative stress induced by ethanol treatment by the decreased expression of SOD at mRNA level, and demonstrates antioxidative neuroprotective effect of CP and SP against developmental ethanol exposure in vitro.

Animals ; Antiviral Agents ; pharmacology ; Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Ethanol ; toxicity ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Hippocampus ; cytology ; embryology ; metabolism ; Isoflavones ; pharmacology ; Male ; Mice ; Mice, Inbred Strains ; Plant Extracts ; pharmacology ; Pregnancy ; Pueraria ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Superoxide Dismutase ; genetics