Kim, Seong-Yeon;Na, Yeon-Joo;Kim, Dong-Ju;Kim, Yeong-Seok;Kim, Hyeong-Min;Hwang, Sung-Ha;Kwak, Ji-Yeon;Kuh, Hyo-Jeong;Lee, Jae-Hwi
The Korean Journal of Physiology and Pharmacology
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v.16
no.3
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pp.205-209
/
2012
The objective of the present study was to establish the method of measurement of hydrogen peroxide and to estimate the anti-oxidative effect of genistein in the skin. UVB induced skin oxidation and anti-oxidative effect of genistein formulations were evaluated by determining levels of hydrogen peroxide. The mechanism involved in the determination of hydrogen peroxide is based on a color reaction between ferric ion ($Fe^{3+}$) and xylenol orange, often called FOX assay and subsequent monitoring of absorbance values of the reactant at 540 nm. The reaction was to some extent pH-dependent and detection sensitivity was greatest at pH 1.75. Genistein liposomal gel demonstrated better anti-oxidative effect with regard to lowering hydrogen peroxide levels elevated by UVB irradiation compared to genistein-suspended gel. A linear relationship has been observed between anti-oxidative effect of genistein and drug deposition in the skin tissue. Genistein liposomal gel resulting in the localization of the drug in the deeper skin led to improved anti-oxidative effect compared to genistein gel. The suggested method for evaluation of oxidation of the skin can be used as a tool to screen effective anti-oxidative agents and their delivery systems acting on the skin.
The aim of this study was to investigate the in vitro antioxidant profiles of genistein and other isoflavonoids. The reactivity of genistein towards stable radical and reactive oxygen species including ${\bullet}\;ABTS^+$, ${\bullet}{O_2}^-$, $H_2O_2$ and HOCl has been investigated, and the effects were compared with other isoflavonoids and antioxidants. All the tested isoflavonoids showed remarkable ${\bullet}\;ABTS^+$ scavenging activity and genistein was more potent than BHT and ascorbic acid. Genistein was more effective in scavenging hypochlorous acid than superoxide and hydrogen peroxide. At $10\;{\mu}M$ concentrations of genistein and genistin showed about 90% inhibitory effect on HOCl, while BHT and ascorbic acid showed lower than 50% inhibitory effect. Moreover, genistein could inhibit plasmid DNA cleavage, protein degradation and cell death from HOCl attack, while daidzein, BHT and ascorbic acid could not protect them effectively. These results suggest that genistein is a more potent radical scavenger than other isoflavonoids, and it can remarkably reduce cellular damage induced by HOCl.
This study was undertaken to determine the bioavailability of isoflavones in weanling Sprague-Dawley rats by providing diets containing different levels of soy isoflavones for 6 weeks: 0.025% (low isoflavone intake; LI), 0.125% (medium isoflavone intake; MI), and 0.25% (high isoflavone intake; HI). The subsequent fecal and urinary excretion of daidzein and genistein was then measured. As the levels of dietary isoflavones increased, the amount of food intakes significantly decreased, and weight gain was slower in female rats. In male rats, there was no significant difference in weight gains related to dietary intakes. Urinary excretion of daidzein and genistein was significantly higher in the MI and HI groups in both male and female rats than the control and LI groups. The recovery % of daidzein and genistein in the urine was significantly lower in the MI and HI groups. Fecal daidzein increased as dietary isoflavone intakes increased in female rats; however, in male rats the increase was significant only in the HI group. The recovery % of daidzein and genistein in the feces of female rats was not significantly different among the four groups. When dietary isoflavones were increased from 0.025% to 0.25%, the amounts of daidzein and genistein excreted in the urine and feces increased; however, the low recovery rate of both daidzein and genistein in the urine implies an increased bioavailability of isoflavones. We also observed sex-related differences in the urinary and fecal recovery of isoflavone intakes.
Julong 601, a Gram-positive anaerobic bacterium strain capable of cleaving the C-ring of isoflavones daidzein and genistein, was isolated from human feces. BLAST search revealed that its complete 16S rDNA gene sequence has 99% similarity to Eubacterium ramulus. Metabolites of daidzein and genistein were determined as O-desmethylangolensin (O-Dma) and 2-(4-hydroxyphenyl) propionic acid (2-HPPA), respectively, based on UV, EI-MS, and $^1H$ and ^{13}C$ NMR spectral analyses. Enantiomers of O-Dma and 2-HPPA were isolated by chiral stationary-phase HPLC (CSP HPLC). Cleavage of the C-ring of daidzein and genistein by strain Julong 601 was highly enantioselective. Specific rotation ([$\alpha]_D$) and circular dichroism (CD) spectra of the enantiomers are reported here for the first time. Biotransformation kinetics of daidzein and genistein indicated that the C-ring of genistein has a higher susceptibility to bacterial degradation than that of daidzein.
Kim, Yoo-Kyeong;Kim, Hyeon-A;Park, Min-Young;Do, Sun-Hee
Preventive Nutrition and Food Science
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v.12
no.1
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pp.11-15
/
2007
This study was conducted to evaluate the influence of diets containing genistein and soy extract on the growth of MDA-MB-231 cells implanted into female Balb/c mice. Four-week-old female athymic nude mice (Balb/c) were acclimated to an AIN-93G control diet for 1 week and then injected MDA-MB-231 cells ($1{\times}10^6$/site) and were continued on the on AIN-93G control diet. Five weeks after injecting the MDA-MB-231 cells ($1{\times}10^6$/site), two experimental groups were assigned to diets containing genistein (750 ${\mu}g/g$ AIN-93G diet) or 0.6% soy extract (containing genistein at 750 ${\mu}g/g$ AIN-93G diet) until they were sacrificed. Tumor growth was significantly reduced in the groups treated with genistein and soy extract compared to the control group. The results of the proliferating cell nuclear antigen (PCNA) assay also revealed that genistein and soy extract treatment reduced the proliferation of MDA-MB-231 cells in vivo. In the present study, dietary isoflavone was provided just before solid tumor formation, and thus the timing of dietary isoflavone administration may be critical to the suppression of tumor growth.
This study was directed to finding an optimum extraction condition of genistein from the S. flavescens with supercritical carbon dioxide as a solvent. In this effort, effects of the extraction conditions including pressure, temperature and a co-solvent on the extraction efficiency were investigated. The aqueous ethanol and methanol solutions were used as co-solvents while the tested operating pressure and temperature ranges were from 200 bar to 300 bar and from 308.15 K to 323.15 K, respectively. The concentration of genistein was determined by means of HPLC equipped with a UV detector. From the results, it was observed that an increase in pressure led to the higher extraction efficiency. Further, methanol showed better performance as a co-slovent than ethanol. The DPPH radical scavenging activities were measured to compare antioxidant activities of S. flavescens extracts.
Lee Young Min;Jung Myeong Ho;Lee Yeon Sook;Song Jihyun
Journal of Nutrition and Health
/
v.37
no.9
/
pp.786-793
/
2004
Women with menopause or rats with ovariectomy is associated with increased body weight, body fat and insulin resistance, which are components of metabolic syndrome. Increased prevalence of metabolic syndrome after menopause might be associated with mitochondrial dysfunction, since mitochondrial oxidative and phosphorylation activity is strongly correlated with insulin sensitivity. Although estradiol replacement prevents the metabolic syndrome, harmful effect of estradiol hampers the casual usage to prevent the metabolic syndrome. It has been reported that genistein has a mild estrogenic activity, decreases fat mass in mice and has an antidiabetic role in diabetic rats. Although insulin resistance is closely related to mitochondrial functions, there has not been yet any study in regard to the effect of dietary genistein on mitochondrial function in the insulin resistant female subjects induced by ovariectomy or similar situation. The present study investigated whether the supplementation of genistein in the high fat diet affected the mitochondrial function of high fat fed ovariectomized rats. Female Sprague Dawley rats (8 weeks old) were assigned to the following groups: sham-operated+ high fat diet (S, n=6); sham-operated + high fat diet with 0.1% genistein (S + G, n=7); ovariectomized + high fat diet (OVX, n=8); ovariectomized + high fat diet with 0.1% genistein (OVX+ G, n=8). Ovariectomy significantly increased body weight compared with S group. Genistein consumption in ovariectomized (OVX + G) rats decreased body weight gain compared with OVX rats. Liver weights were increased by ovariectomy. The hepatic mitochondrial protein density expressed as mg per g liver was lower in the OVX group than in the S group. However, OVX + G group showed the increased mitochondrial protein density similar to the level of S group. When mRNA levels of genes related to mitochondria such as peroxisome proliferator-activated receptor ${\gamma}$ coactivator 1 (PGC-1) and cytochrome c oxidase subunit III (COX III) were measured, there were decreases in the mRNA levels of PGC-1 and COX III in S + G, OVX and OVX + G group. The activity of cytochrome c oxidase was not different between groups. We could observe the decrease in succinate dehydrogenase (SDH) activity per g liver in OVX rats. Genistein supplement increased SDH activity. In conclusion, genistein supplementation to the OVX rats enhanced mitochondrial function by increasing mitochondrial protein density and SDH activity. The improvement in mitochondrial function by genistein can contribute to the improvement in metabolic syndrome.
Aspirin and its deacetylated form, sodium salicylate (NaSal), have been shown to exert chemopreventive activities against many human cancers including those of the colon, lung, and breast. Previously, we showed that combined treatment of NaSal and genistein synergistically induced apoptosis in A549 lung cancer cells, indicating that these two natural chemicals could be used in combination for cancer therapy. In this study, we examined effects of NaSal/genistein combined treatment on other cancer cells and in three-dimensional multicellular tumor spheroid (MTS) and in an in vitro solid tumor model. We found that the combined treatment induces apoptosis in the HCT116 cells and the A549 cells, but not in the MCF-7 cells. Interestingly, the MCF-7 cells responded to the NaSal/genistein combined treatment by undergoing cell death when they were cultivated as MTS. The combined treatment induced apoptosis at an earlier stage in the MCF-7 MTS culture. However, when the MCF-7 MTS was cultivated for a longer period, it induced necrosis rather than apoptosis. We further found that the apoptotic pattern observed in MCF-7 MTS was incomplete: the chromatins were condensed and fragmented, but the nuclear membrane was still intact. Taken together, these results demonstrate that the NaSal/genistein combined treatment induces incomplete apoptosis and necrosis in the MCF-7 MTS culture system.
Radiotherapy is currently applied in the treatment of human cancers. We studied whether genistein would enhance the radiosensitivity and explored its precise molecular mechanism in cervical cancer cells. After co-treatment with genistein and irradiation, the viability, cell cycle analysis, and apoptosis signaling cascades were elucidated in CaSki cells. The viability was decreased by co-treatment with genistein and irradiation compared with irradiation treatment alone. Treatment with only ${\gamma}$-irradiation led to cell cycle arrest at the $G_1$ phase. On the other hand, co-treatment with genistein and ${\gamma}$-irradiation caused a decrease in the $G_1$ phase and a concomitant increase up to 56% in the number of $G_2$ phase. In addition, co-treatment increased the expression of p53 and p21, and Cdc2-tyr-15-p, supporting the occurrence of $G_2/M$ arrest. In general, apoptosis signaling cascades were activated by the following events: release of cytochrome c, upregulation of Bax, down regulation of Bcl-2, and activation of caspase-3 and -8 in the treatment of genistein and irradiation. Apparently, co-treatment downregulated the transcripts of E6*I, E6*II, and E7. Genistein also stimulated irradiation-induced intracellular reactive oxygene, species (ROS) production, and co-treatment-induced apoptosis was inhibited by the antioxidant N-acetylcysteine, suggesting that apoptosis has occurred through the increase in ROS by genistein and ${\gamma}$-irradiation in cervical cancer cells. Gamma-irradiation increased cyclooxygenase-1 (COX-2) expression, whereas the combination with genistein and ${\gamma}$-irradiation almost completely prevented irradiation-induced COX-2 expression and $PGE_2$ production. Co-treatment with genistein and ${\gamma}$-irradiation inhibited proliferation through $G_2/M$ arrest and induced apoptosis via ROS modulation in the CaSki cancer cells.
Kim, Tae-Hee;Yuh, In-Suh;Park, In-Chul;Cheong, Hee-Tae;Kim, Jong-Taek;Park, Choon-Keun;Yang, Boo-Keun
Journal of Embryo Transfer
/
v.29
no.2
/
pp.141-148
/
2014
Quercetin and genistein, plentifully present in fruits and vegetables, are flavonoid family members that have antioxidative function and plant-derived phytoestrogen activity. The antioxidative effects of quercetin and genistein on boar sperm characteristics and in vitro development of IVF embryo were investigated. The sperm motility was increased by addition of genistein $50{\mu}M$ for 6 hr incubation compared to control (p<0.05). The sperm viability was increased by addition of quercetin 1 and $50{\mu}M$ and genestein 1 and $50{\mu}M$ for 3 hr incubation. In addition, the sperm viability seemed to be increased dose-dependantly by addition of quercetin or genistein 1 and $50{\mu}M$, respectively (p<0.05). The membrane integrities were not increased by quercetin or genistein treatments for 3 hr or 6 hr incubation period except for quercetin $1{\mu}M$ for 3 hr incubation. In mitochondrial activities, addition of quercetin $50{\mu}M$ for 6 hr incubation increased mitochondrial activity but decreased at $100{\mu}M$ concentration compared with control (p<0.05). When porcine IVF embryos were cultured in PZM-3 medium supplemented with low concentrations of quercetin ($1{\sim}10{\mu}M$), the developmental rates to morula and blastocyst increased but significantly decreased at high concentrations of quercetin ($25{\sim}50{\mu}M$). The highest developmental rate to blastocysts among all concentrations of quercetin was shown at quercetin $10{\mu}M$ (p<0.05). The developmental rates to morula or blastocysts at low ($0.01{\sim}1{\mu}M$) and high ($5{\sim}10{\mu}M$) concentrations of genistein were not significantly different among all treatment group and genistein did not affect on IVF embryo development. These results suggest that quercetin and genistein seem to have positive effects at certain concentrations on sperm characteristics such as motility, viability and mitochondrial activity. In addition, low concentrations of quercetin (1, 5 and $10{\mu}M$) in this experiment, seem to have beneficial effect on porcine IVF embryo development but genistein did not affect on it at all given concentrations ($0.01{\sim}10{\mu}M$).
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