Studies on the antioxidant properties of some phytoestrogens

Isoflavones genistein and daidzein are nonsteroidal phytoestrogens occurring mainly in soybean foods. These phytoestrogens possess estrogenic properties and show a variety of health benefits as anti‐inflammatory agents. However, the mechanism of their action has not been identified in detail. The aim of this study is to characterize the antioxidant powers of genistein, daidzein and daidzein metabolite–equol through their activities to scavenge superoxide anion radical (O^?₂^?), hydroxyl radical (HO^?), 2,2–diphenyl–1‐picrylhydrazyl radical (DPPH^?) and hydrogen peroxide (H₂O₂) using chemiluminescence and spectrophotometry techniques. Potassium superoxide in dimethyl sulphoxide (DMSO) and 18‐crown‐6 ether were used as a source of O^?₂^?. Hydroxyl radicals were produced using the Fenton reaction. In free radical assays, genistein had the IC₅₀ values (an amount of antioxidant concentration required to decrease the initial radical concentration by 50%) 0.391 ± 0.012 mM for O^?₂^?, 0.621 ± 0.028 mM for HO^? and 1.89 ± 0.16 mM for DPPH^?. The IC₅₀ values for daidzein for these free radicals were 1.924 ± 0.011 mM, 0.702 ± 0.012 mM and 2.81 ± 0.03 mM, respectively. Equol was the most active the free radical scavenger with IC₅₀ = 0.451 ± 0.018 mM for HO^? and IC₅₀ = 1.36 ± 0.11 mM for DPPH^?. All tested compounds exerted a significant effect on the H₂O₂: IC₅₀ = 18.1 ± 1.1 μM for genistein, IC₅₀ = 2.1 ± 0.5 μM for daidzein, and IC₅₀ = 1.06 ± 0.2 μM for equol. These findings show that genistein, daidzein and equol are effective free radical scavengers and possess high antioxidant power in vitro. Copyright © 2016 John Wiley & Sons, Ltd.     

雌马酚产生细菌及其雌马酚合成代谢机制

大豆食品中通常富含染料木素和大豆苷元等异黄酮素,人和动物肠道中的某些细菌具有将异黄酮素代谢转化为S-雌马酚的能力。到目前为止,S-雌马酚被认为是一种具有潜在健康调节作用的化合物。啮齿类动物均具备产雌马酚的能力,但不同人群之间存在差异,产雌马酚细菌是否存在可能是造成这种差异的重要原因;不同产雌马酚细菌的代谢机制可能不同,并影响机体最终产雌马酚的能力。本文对已知的各种产雌马酚细菌及其细菌的雌马酚合成机制进行综述,以期为进一步了解雌马酚产生个体差异、雌马酚代谢转化效率、体外雌马酚的发酵生产,以及临床产雌马酚细菌的应用等提供理论参考。     

Flavonoid metabolism: the interaction of metabolites and gut microbiota

Abstract

Several dietary flavonoids exhibit anti-oxidative, anti-inflammatory, and anti-osteoporotic activities relevant to prevention of chronic diseases, including lifestyle-related diseases. Dietary flavonoids (glycoside forms) are enzymatically hydrolyzed and absorbed in the intestine, and are conjugated to their glucuronide/sulfate forms by phase II enzymes in epithelial cells and the liver. The intestinal microbiota plays an important role in the metabolism of flavonoids found in foods. Some specific products of bacterial transformation, such as ring-fission products and reduced metabolites, exhibit enhanced properties. Studies on the metabolism of flavonoids by the intestinal microbiota are crucial for understanding the role of these compounds and their impact on our health. This review focused on the metabolic pathways, bioavailability, and physiological role of flavonoids, especially metabolites of quercetin and isoflavone produced by the intestinal microbiota.     

A combination of soy isoflavones and cello-oligosaccharides changes equol/O-desmethylangolensin production ratio and attenuates bone fragility in ovariectomized mice

We examined the cooperative effects of isoflavones and cello-oligosaccharides on daidzein metabolism and bone fragility in ovariectomized mice. Cello-oligosaccharides increased urinary equol and decreased O-desmethylangolensin. A combination of isoflavones and cello-oligosaccharides attenuated decreases in bone breaking force and stiffness caused by ovariectomy. Combination treatment with isofalvones and cello-oligosaccharides increases urinary equol/O-desmethylangolensin production ratio and prevents ovariectomy-induced abnormalities in bone strength.     

雌马酚的生物活性和临床应用

雌马酚是大豆异黄酮(SI)的主要组分之一——大豆素(Dai)的代谢产物。雌马酚较其原型具有更为有效的生物学作用,虽其作用及机制还存在争议,但很多研究表明雌马酚的生物学作用以及在适用人群显然都优于SI,并受到普遍关注。研究和开发雌马酚的生物活性,在多种常见慢性病的预防与控制中有重要的理论和实际意义。     

The effect of equol injection in ovo on posthatch growth, meat quality and antioxidation in broilers

In order to investigate the long-term effects of equol (Eq) on growth and meat quality in broilers, 0 μg (control, Con), 20 μg (low dose, L) and 100 μg (high dose, H) Eq, respectively, were injected into fertile eggs (146 eggs per group) on 7 days of embryos. After hatch, chickens were fed under the same conditions and slaughtered at 49 days of age for sample collection and analysis. The results showed that body weight and composition were marginally affected by Eq administration (P > 0.05). Compared with their male counterparts, the meat quality of female broilers was affected greatly after Eq administration. The redness (a*) of meat color in the L and H groups of female broilers was significantly decreased by 24.10% and 21.50% (P < 0.01), respectively; cooking loss decreased by 12.11% and 16.82%, respectively, in the L and H groups (P < 0.01); 24 h and 48 h drip loss was significantly decreased by 60.27% and 45.72% (P < 0.05), respectively, in the H group. However, for male broilers, only cooking loss was significantly decreased by high dosage of Eq treatment (P < 0.05). The antioxidative status was analyzed for discovering further the mechanism behind the improvement of the water-holding capacity caused by Eq in female broilers. The activity of glutathione peroxidase (GSHPx) in plasma was greatly increased by 15.94% in the L group (P < 0.01), whereas the total superoxide dismutase activity (T-SOD) and the content of malondialdehyde in plasma were not changed (P > 0.05). The T-SOD activity in the breast muscle of the L and H groups were significantly improved by 23.14% and 18.82% (P < 0.05), respectively. GSHPx in the breast muscle of the H group showed a tendency to increase (P = 0.06 < 0.1). These results indicate that Eq injection in ovo does not affect the growth of broilers, but significantly improves the water-holding capacity of the muscle, especially in female broilers, which is related to the improvement of antioxidative status.     

Performance and egg quality of laying hens fed flaxseed: highlights on n-3 fatty acids,cholesterol, lignans and isoflavones

Flaxseed is a rich source of α-linolenic acid and phytoestrogens, mainly lignans, whose metabolites (enterodiol and enterolactone) can affect estrogen functions. The present study evaluated the influence of dietary flaxseed supplementation on reproductive performance and egg characteristics (fatty acids, cholesterol, lignans and isoflavones) of 40 Hy-Line hens (20/group) fed for 23 weeks a control diet or the same diet supplemented with 10% of extruded flaxseed. The flaxseed diet had approximately three times the content of lignans (2608.54 ng/g) as the control diet, mainly secoisolariciresinol diglucoside (1534.24 v. 494.72 ng/g). When compared with the control group, hens fed flaxseed showed a similar deposition rate (72.0% v. 73.9%) and egg yield. Furthermore, there was no effect of flaxseed on the main chemical composition of the egg and on its cholesterol content. Estradiol was higher in the plasma of the control group (1419.00 v. 1077.01 pg/ml) probably due to the effect of flaxseed on phytoestrogen metabolites. The plasma lignans were higher in hens fed flaxseed, whereas isoflavones were lower, mainly due to the lower equol value (50.52 v. 71.01 ng/ml). A similar trend was shown in eggs: the flaxseed group had higher level of enterodiol and enterolactone, whereas the equol was lower (198.31 v. 142.02 ng/g yolk). Secoisolariciresinol was the main lignan in eggs of the flaxseed group and its concentration was three times higher then control eggs. Flaxseed also improved the n-3 long-chain polyunsaturated fatty acids of eggs (3.25 v. 0.92 mg/g egg), mainly DHA, however, its oxidative status (thiobarbituric reactive substances) was negatively affected. In conclusion, 10% dietary flaxseed did not affect the productive performance of hens or the yolk cholesterol concentration, whereas the lignans and n-3 polyunsaturated fatty acid content of eggs improved. Further details on the competition between the different dietary phytoestrogens and their metabolites (estrogen, equol, enterodiol and enterolactone) should be investigated.     

Evaluation of inter‐individual differences in gut bacterial isoflavone bioactivation in humans by PCR‐based targeting of genes involved in equol formation

Aim

To identify human subjects harbouring intestinal bacteria that bioactivate daidzein to equol using a targeted PCR‐based approach.

Methods and Results

In a pilot study including 17 human subjects, equol formation was determined in faecal slurries. In parallel, faecal DNA was amplified by PCR using degenerate primers that target highly conserved regions of dihydrodaidzein reductase and tetrahydrodaidzein reductase genes. PCR products of the expected size were observed for six of the eight subjects identified as equol producers. Analysis of clone libraries revealed the amplification of sequences exclusively related to Adlercreutzia equolifaciens in four of the subjects tested positive for equol formation, whereas in three of the equol producers, only sequences related to Slackia isoflavoniconvertens were observed. No amplicons were obtained for one equol‐forming subject, thus suggesting the presence of nontargeted alternative genes. Amplicons were only sporadically observed in the nonequol producers.

Conclusion

The majority of human subjects who produced equol were also detected with the developed PCR‐based approach.

Significance and Impact of the Study

The obtained results shed light on the distribution and the diversity of known equol‐forming bacterial species in the study group and indicate the presence of as yet unknown equol‐forming bacteria.