Long-term supplementation of genistein improves immune homeostasis in the aged gut and extends the laying cycle of aged laying hens.

Aging is associated with alterations in gut function, including intestinal inflammation, leaky gut, and impaired epithelial regeneration. Rejuvenating the aged gut is imperative to extend the laying cycle of aged laying hens. Genistein is known to have beneficial effects on age-related diseases, but its precise role in homeostasis of the aged gut of laying hens remains to be elucidated. In this study, 160 45-wk-old Hyline Brown laying hens were continuously fed a basal diet or a diet supplemented with 40 mg/kg genistein until they reached 100 wk of age. The results revealed that long-term genistein supplementation led to an improvement in the egg production rate and feed conversion ratio, as well as an increase in egg quality. Moreover, the expression levels of senescence markers, such as ß-galactosidase, P16, and P21, were decreased in the gut of genistein-treated aged laying hens. Furthermore, genistein ameliorated gut dysfunctions, such as intestinal inflammation, leaky gut, and impaired epithelial regeneration. Treg cell-derived IL-10 plays a crucial role in the genistein-induced regulation of age-related intestinal inflammation. This study demonstrates that long-term consumption of genistein improves homeostasis in the aged gut and extends the laying cycle of aged laying hens. Moreover, the link between genistein and Treg cells provides a rationale for dietary intervention against age-associated gut dysfunction.

Genistein transfersome-embedded topical delivery system for skin melanoma treatment: in vitro and ex vivo evaluations.

Skin melanoma is considered the most dangerous form of skin cancer due to its association with high risk of metastasis, high mortality rate and high resistance to different treatment options. Genistein is a natural isoflavonoid with known chemotherapeutic activity. Unfortunately, it has low bioavailability due to its poor aqueous solubility and excessive metabolism. In the current study, genistein was incorporated into transferosomal hydrogel to improve its bioavailability. The prepared transferosomal formulations were characterized regarding: particle size; polydispersity index; zeta potential; encapsulation efficiency; TEM; FTIR; DSC; XRD; in vitro drug release; viscosity; pH; ex vivo anti-tumor activity on 3D skin melanoma spheroids and 1-year stability study at different storage temperatures. The optimized formulation has high encapsulation efficiency with an excellent particle size that will facilitate its penetration through the skin. The transfersomes have a spherical shape with sustained drug release profile. The anti-tumor activity evaluation of genistein transfersome revealed that genistein is a potent chemotherapeutic agent with enhanced penetration ability through the melanoma spheroids when incorporated into transfersomes. Stability study results demonstrate the high physical and chemical stability of our formulations. All these outcomes provide evidence that our genistein transferosomal hydrogel is a promising treatment option for skin melanoma.

Synergistic effect of genistein and adiponectin reduces fat deposition in chicken hepatocytes by activating the ERß-mediated SIRT1-AMPK signaling pathway.

Dietary supplementation with bioactive substances that can regulate lipid metabolism is an effective approach for reducing excessive fat deposition in chickens. Genistein (GEN) has the potential to alleviate fat deposition; however, the underlying mechanism of GEN's fat-reduction action in chickens remains unclear. Therefore, the present study aimed to explore the underlying mechanism of GEN on the reduction of fat deposition from a novel perspective: intercellular transmission of adipokine between adipocytes and hepatocytes. The findings showed that GEN enhanced the secretion of adiponectin (APN) in chicken adipocytes, and the enhancement effect of GEN was completely blocked when the cells were pretreated with inhibitors targeting estrogen receptor ß (ERß) or proliferator-activated receptor γ (PPARγ) signals, respectively. Furthermore, the results demonstrated that both co-treatment with GEN and APN or treatment with the medium supernatant (Med SUP) derived from chicken adipocytes treated with GEN significantly decreased the content of triglyceride and increased the protein levels of ERß, Sirtuin 1 (SIRT1) and phosphor-AMP-activated protein kinase (p-AMPK) in chicken hepatocytes compared to the cells treated with GEN or APN alone. Moreover, the increase in the protein levels of SIRT1 and p-AMPK induced by GEN and APN co-treatment or Med SUP treatment were blocked in chicken hepatocytes pretreated with the inhibitor of ERß signals. Importantly, the up-regulatory effect of GEN and APN co-treatment or Med SUP treatment on the protein level of p-AMPK was also blocked in chicken hepatocytes pretreated with a SIRT1 inhibitor; however, the increase in the protein level of SIRT1 induced by GEN and APN co-treatment or Med SUP treatment was not reversed when the hepatocytes were pretreated with an AMPK inhibitor. In conclusion, the present study demonstrated that GEN enhanced APN secretion by activating the ERß-Erk-PPARγ signaling pathway in chicken adipocytes. Subsequently, adipocyte-derived APN synergized with GEN to activate the ERß-mediated SIRT1-AMPK signaling pathway in chicken hepatocytes, ultimately reducing fat deposition. These findings provide substantial evidence from a novel perspective, supporting the potential use of GEN as a dietary supplement to prevent excessive fat deposition in poultry.

Effects of the phytoestrogen genistein on the development of the reproductive system of Sprague Dawley rats

OBJECTIVES: Genistein is known to influence reproductive system development through its binding affinity for estrogen receptors. The present study aimed to further explore the effect of Genistein on the development of the reproductive system of experimental rats. METHODS: Eighteen post-weaning female Sprague Dawley rats were divided into the following groups: (i) a control group that received vehicle (distilled water and Tween 80); (ii) a group treated with 10 mg/kg body weight (BW) of Genistein (Gen 10); and (iii) a group treated with a higher dose of Genistein (Gen 100). The rats were treated daily for three weeks from postnatal day 22 (P22) to P42. After the animals were sacrificed, blood samples were collected, and the uteri and ovaries were harvested and subjected to light microscopy and immunohistochemical study. RESULTS: A reduction of the mean weekly BW gain and organ weights (uteri and ovaries) were observed in the Gen 10 group compared to the control group; these findings were reversed in the Gen 100 group. Follicle stimulating hormone and estrogen levels were increased in the Gen 10 group and reduced in the Gen 100 group. Luteinizing hormone was reduced in both groups of Genistein-treated animals, and there was a significant difference between the Gen 10 and control groups (p<0.05). These findings were consistent with increased atretic follicular count, a decreased number of corpus luteum and down-regulation of estrogen receptors-a in the uterine tissues of the Genistein-treated animals compared to the control animals. CONCLUSION: Post-weaning exposure to Genistein could affect the development of the reproductive system of ovarian-intact experimental rats because of its action on the hypothalamic-pituitary-gonadal axis by regulating hormones and estrogen receptors.

Efeitos de altas doses de genisteína sobre o epitélio mamário de ratas / Effects of high doses of genistein on mammary gland of female rat

OBJETIVO: avaliar os efeitos de altas doses de genisteína sobre o epitélio mamário de ratas adultas. MÉTODOS: após 28 dias da ooforectomia, cinquenta ratas adultas foram divididas em cinco grupos, a saber: um controle (Ctrl), três que receberam genisteína (GEN) nas doses de 46 mg/kg (GEN46), 125 mg/kg (GEN125) e 250 mg/kg (GEN250), e um que recebeu estrogênios conjugados equinos na dose de 50 µg/kg (ECE). As substâncias foram administradas diariamente durante 30 dias consecutivos por gavagem e na última semana de tratamento foi efetuado exame colpocitológico durante sete dias consecutivos. Após o tratamento, os animais foram anestesiados, amostras de sangue foram retiradas para determinação do estradiol e da progesterona, e o primeiro par de mamas inguinais retirado e processado para análise histomorfométrica. Os dados obtidos foram submetidos à análise de variância complementada pelo teste de Tukey-Kramer (p<0,05). RESULTADOS: nos grupos Ctrl e tratados com as diferentes doses de GEN as mamas apresentaram-se atróficas, no entanto mostraram-se desenvolvidas no grupo ECE, onde se notou a presença de inúmeros ductos e alvéolos mamários contendo material eosinófilo em seu interior. A morfometria mostrou maior área de parênquima mamário no grupo ECE (98.870,1±550,4 µm²* por mm²; p<0,05) comparado aos outros grupos (Ctrl=36.875,6±443,4; GEN46=37.001,7±557,4; GEN125=36.480,8±658,3 e GEN250=37.502,8±669,3). O mesmo ocorreu em relação ao número de alvéolos e ductos mamários no grupo ECE (33,2±6,9* por mm²; p<0,05) em relação aos outros grupos (Ctrl=10,4±2,1, GEN 46=11,2±3,1; GEN 125=11,6±2,1 e GEN 250=12,3±2,3). Os níveis de estradiol mostraram-se aumentados no grupo ECE em relação aos outros grupos (9,4±1,7 pg/mL; p<0,05), sendo que os níveis séricos de progesterona mostraram-se semelhantes em todos os grupos de estudo. CONCLUSÃO: a administração de genisteína em altas doses não apresentou efeito proliferativo no tecido mamário de ratas.

PURPOSE: to evaluate the effects of high doses of genistein on the mammary glands of adult female rats. METHODS: Twenty-eight days after oophorectomy, 50 adult female rats were divided into five groups, as follows: a control group (Ctrl), three rats that received genistein (GEN) at the doses of 46 mg/kg (GEN46;), 125 mg/kg (GEN125) and 250 mg/kg (GEN250); one group received conjugated equine estrogen at the dose of 50 µg/g (ECE50). The substances were administered daily for 30 consecutive days by gavage and in the last week of the period of treatment, colpocytological exams were carried out for seven consecutive days. After treatment, the animals were anesthetized, blood samples were collected for estradiol and progesterone determination and the first pair of inguinal mammary glands was removed and processed for histomorphometric analysis. Collected data were subjected to analysis of variance supplemented by the Tukey-Kramer test (p<0.05). RESULTS: the ctrl group and the ones treated with different doses of GEN showed atrophic mammary glands, whereas the glands were more developed in the ECE group, where numerous mammary ducts and alveoli were observed. Morphometry showed a larger area of mammary parenchyma in the ECE group (98.870.1±550.4 µm²* per mm²; p<0.05) compared with other groups (Ctrl=36.875.6±443.4; GEN46=37.001.7±557.4; GEN125=36.480.8±658.3 and GEN250=37.502.8±669.3). The same occurred in the number of alveoli in the ECE group (33.2±6.9* per mm²; p<0.05) compared to the other groups (Ctrl=10.4±2.1, GEN46=11.2±3.1; GEN125=11.6±2.1 and GEN250=12.3±2.3). The estradiol level was higher in the ECE group compared to the other groups (9.4±1.7 pg/mL; p<0.05), whereas serum levels of progesterone were similar in all groups. CONCLUSION: the administration of genistein at high doses had no trophic effect on the mammary glands of rats.

Efeitos das isoflavonas em altas doses sobre o útero da rata / Effects of high-dose isoflavones on rat uterus

OBJETIVO: Avaliar o efeito de altas doses de isoflavonas no útero de ratas adultas castradas. MÉTODOS: Ratas virgens ovariectomizadas (n = 40) foram tratadas por 30 dias consecutivos com veículo (GCtrl) ou genisteína nas concentrações 42 (GES42), 125 (GES125) e 250 (GES250) µg/g de peso corporal ao dia. O extrato de soja e o veículo (propilenoglicol) foram administrados por gavagem. Ao final do experimento, foi realizada dosagem sérica de 17 β-estradiol e progesterona, avaliou-se o peso dos animais e dos úteros e foi feito exame colpocitológico. Fragmentos do terço médio dos cornos uterinos foram fixados em formol a 10 por cento e processados para inclusão em parafina para estudo histológico. Cortes de 5 µm de espessura foram corados pelo HE e destinados a estudo em microscopia de luz. Analisou-se a histomorfologia do endométrio, área endometrial, número e área ocupada pelas glândulas, assim como a concentração de eosinófilos presentes na lâmina própria. Os dados numéricos obtidos foram submetidos à análise de variância complementada pelo teste de Tukey-Kramer (p < 0,05). RESULTADOS: Verificou-se que o peso do útero, as áreas endometrial e glandular e o número de glândulas e de eosinófilos foram maiores nos animais dos grupos GES250 > GES125 do que nos outros grupos (GES250 > GES125 > GES42 = GCtrl; p < 0,05). Os dados morfológicos mostraram proliferação endometrial nos grupos ES125 e ES250, que apresentavam endométrio mais desenvolvido que outros grupos. Em todos os animais do grupo ES250 notou-se a presença de metaplasia escamosa. CONCLUSÃO: A administração de isoflavonas em altas doses promove metaplasia escamosa no endométrio.

OBJECTIVE: To evaluate the effects of high-dose isoflavones on the uterus of castrated adult rats. METHODS: Adult, ovariectomized virgin rats (n = 40) were treated by gavage during 30 consecutive days with vehicle (propylene glycol, group GCtrl) or different doses of genistein: 42 (group GES42), 125 (GES125), or 250 (GES250) µg/g body weight per day. Animals were killed, weighed, vaginal and uterine samples were taken for cytologic evaluation, and serum levels of 17 β-estradiol and progesterone were determined. The middle third of the uterine horns was dissected, fixed in 10 percent formaldehyde and processed for paraffin inclusion; 5-µm thick sections were obtained and stained with HE for further histological study under light microscopy. The endometrial morphology and area, number and area of glands, and number of eosinophils in the lamina propria were analyzed. ANOVA and the Tukey-Kramer test were used for statistical analyses. RESULTS: Uterine weight, endometrial glandular area, and number of glands and eosinophils were all higher in GES250 > G125 than in the other groups (GES250 > GES125 > GES42 = GCtrl; p < 0.05). Morphological data showed signs of endometrial proliferation upon treatment with genistein, especially in animals in GES125 and GES250 compared to other groups. In all animals in GES250, signs of uterine squamous metaplasia were observed. CONCLUSION: A short treatment period with high daily doses of isoflavones can promote endometrial squamous metaplasia in ovariectomized rats.

Ação das isoflavonas no estresse oxidativo na pós-menopausa da mulher: [revisão] / Effect of isoflavones on oxidative stress in postmenopausal woman: [review]

O presente trabalho apresenta uma revisão bibliográfica da ação das isoflavonas como agentes antioxidantes. Refere que a geração de radicais reativos ao oxigênio dentro da célula faz parte do metabolismo celular, o que culmina no desenvolvimento de mecanismos de defesa antioxidantes para a retirada desses radicais. Inúmeros trabalhos referem que os asiáticos apresentam menor índice de tumores mamários, de endométrio e de próstata, o que pode estar correlacionado com o mecanismo da ação das isoflavonas, tanto como moduladores seletivos dos receptores hormonais (SERMs) quanto por sua ação antioxidante. No entanto, procuramos chamar a atenção para sua ação antioxidante.

This paper presents a brief review of the action of isoflavones as antioxidants. Being the generation of free radicals within the cell a part of the cell metabolism, evolution predictably developed efficient antioxidant defense mechanisms to remove these radicals. Numerous studies report that Asians have lower rates of breast tumors, endometrial and prostate, a fact which could be postulated to correlate with the moderate to high daily intake of isoflavones-containing foods by these human groups. The effects of isoflavones significantly lowering cancer rates can be accounted for not only by their antioxidant action but by their action as selective hormone receptor modulators (SERMs) as well. This review focuses on the antioxidantaction of isoflavones.