Doses Of Genistein Research Articles

Effect of vehicle on the in vitro penetration and metabolism of genistein and daidzein in ex vivo skin explants and the Phenion full-thickness skin model.

In a read-across assessment of the safety of genistein and daidzein in cosmetic products, additional information was required to account for differences in their systemic exposure after topical application in a typical body lotion formulation. Therefore, we measured the penetration and metabolism of two doses (3 and 30 nmoles/cm2) of genistein and daidzein applied in ethanol and in a body formulation to fresh pig skin, fresh and frozen human skin, and PhenionFT models. Both chemicals readily penetrated all skin models when applied in ethanol. The same sulfate and glucuronide metabolites were formed in fresh pig skin, fresh human skin, and PhenionFT models, which also all demonstrated that (a) these pathways could be saturated between 3 and 30 nmoles/cm2 and (b) the extent of metabolism of daidzein was lower than genistein. Although the relative amounts of radiolabeled chemical in human skin and medium compartments were altered by freezing, their overall bioavailability was not affected. The greatest impact on the bioavailability and distribution of both chemicals was observed when they were applied in the formulation. Most of the dose applied in the formulation was retained on the skin surface, especially at 30 nmoles/cm2 (60%-90%), resulting in much lower amounts in the medium and/or skin. In conclusion, all four skin models demonstrated first-pass metabolism of genistein and daidzein and a marked alteration in their disposition by applying them in a body lotion formulation. This supports the use of fresh pig skin and PhenionFT models as alternatives to human skin for investigating skin metabolism and formulation effects for these two chemicals. The results were used to develop the dermal module of a PBPK model and dose setting for organ-on-chip experiments. They could also be used to refine internal exposure estimates in regulatory safety assessments.

Restoration of Radiosensitivity by Soya Isoflavone Genistein is Accomplished by Facilitating DNA Damage Response in Radioresistant Cervical Cancer in vitro

ABSTRACT Context: Enhancing radiotherapeutic efficacy in tumor cells and sparing the normal tissues are major clinical concerns for the betterment of cancer therapy. Genistein (GEN) being a radiosensitizer ameliorates the effectiveness of radiation-induced cell killing by inducing DNA damage. This molecule is accountable for minimizing radiation-related toxicity and protecting healthy cells. However, the explicit mechanism of action of such molecules needs exploration. Aims: The objective of this study is to investigate the mechanistic action of GEN in cervical cancer cell radiosensitization. Settings and Design: Cervical squamous carcinoma cell SiHa and a radioresistant subline SiHa/RR (developed and isolated from SiHa) were taken for this study. The experiments were performed by pretreating the cells with IC30 dose of GEN, followed by acute irradiation to detect the impact of GEN in imparting radiosensitivity. Subjects and Methods: Optimal dose selection of GEN was performed by MTT assay, and radiosensitizing potency was determined by pretreating the cells with IC30 dose of GEN, followed by challenging with acute incremental doses of radiation. Mechanistic parameters were checked by clonogenic assay, cell cycle analysis, DNA damage estimation, apoptosis, and wound healing-sphere-forming assay. Statistical Analysis Used: Statistical analysis was performed in GraphPad software by performing the Student’s t-test. Results: Results depicted decreased numbers of colonies, increased frequency of DNA damage and apoptotic cells, and suppressed wound healing ability along with restrained sphere-forming ability upon the intervention of cells with GEN before radiation exposure. Such observations implied that GEN pretreatment renders improved radiosensitivity in cervical cancer by increased DNA damage-mediated G2/M arrest with subsequent apoptosis. Conclusions: GEN by inducing DNA damage stimulates radiation-induced cell killing in vitro.

Next-generation risk assessment read-across case study: application of a 10-step framework to derive a safe concentration of daidzein in a body lotion.

Introduction: We performed an exposure-based Next Generation Risk Assessment case read-across study using New Approach Methodologies (NAMs) to determine the highest safe concentration of daidzein in a body lotion, based on its similarities with its structural analogue, genistein. Two assumptions were: (1) daidzein is a new chemical and its dietary intake omitted; (2) only in vitro data were used for daidzein, while in vitro and legacy in vivo data for genistein were considered. Methods: The 10-step tiered approach evaluating systemic toxicity included toxicokinetics NAMs: PBPK models and in vitro biokinetics measurements in cells used for toxicogenomics and toxicodynamic NAMs: pharmacology profiling (i.e., interaction with molecular targets), toxicogenomics and EATS assays (endocrine disruption endpoints). Whole body rat and human PBPK models were used to convert external doses of genistein to plasma concentrations and in vitro Points of Departure (PoD) to external doses. The PBPK human dermal module was refined using in vitro human skin metabolism and penetration data. Results: The most relevant endpoint for daidzein was from the ERα assay (Lowest Observed Effective Concentration was 100 ± 0.0nM), which was converted to an in vitro PoD of 33nM. After application of a safety factor of 3.3 for intra-individual variability, the safe concentration of daidzein was estimated to be 10nM. This was extrapolated to an external dose of 0.5μg/cm2 for a body lotion and face cream, equating to a concentration of 0.1%. Discussion: When in vitro PoD of 33nM for daidzein was converted to an external oral dose in rats, the value correlated with the in vivo NOAEL. This increased confidence that the rat oral PBPK model provided accurate estimates of internal and external exposure and that the in vitro PoD was relevant in the safety assessment of both chemicals. When plasma concentrations estimated from applications of 0.1% and 0.02% daidzein were used to calculate bioactivity exposure ratios, values were >1, indicating a good margin between exposure and concentrations causing adverse effects. In conclusion, this case study highlights the use of NAMs in a 10-step tiered workflow to conclude that the highest safe concentration of daidzein in a body lotion is 0.1%.

Genistein promotes cartilage repair and inhibits synovial inflammatory response after anterior cruciate ligament transection in rats by regulating the Wnt/β-catenin axis.

To confirm the protective mechanism of genistein on osteoarthritis (OA). Firstly, we constructed an anterior cruciate ligament transection (ACLT) rat model and administered two doses of genistein via gavage. The effects of the drug on cartilage damage repair and synovitis in OA rats were evaluated through pain-related behavioral assessments, pathological staining, detection of inflammatory factors, and western blot analysis. Secondly, we constructed IL-1-induced chondrocytes and synovial fibroblast models, co-incubated them with genistein, and evaluated the protective effects of genistein on both types of cells through cell apoptosis and cytoskeleton staining. To verify the role of this pathway, we applied the GSK3β inhibitor TWS119 and the Wnt/β-catenin inhibitor XAV939 to ACLT rats and two types of cells to analyze the potential mechanism of genistein's action on OA. Our results confirmed the protective effect of genistein on joint cartilage injury in ACLT rats and its alleviating effect on synovitis. The results of cell experiments showed that genistein can protect IL-1β-induced chondrocytes and synovial fibroblasts, inhibit IL-1β-induced cell apoptosis, increase the fluorescence intensity of F-actin, and inhibit inflammatory response. The results of in vivo and in vitro mechanism studies indicated that TWS119 and XAV939 can attenuate the protective effects of genistein on OA rats and IL-1-induced cell damage. Our research confirmed that genistein may be an effective drug for treating osteoarthritis. Furthermore, we discussed and confirmed that the GSK3β/Wnt/β-catenin axis serves as a downstream signaling pathway of genistein, providing theoretical support for its application.

In An Incisional Model of Wound Healing, Genistein Aglycone Enhances Skin Recovery: A Comparison with Raloxifene and Estradiol in Ovariectomized Rats Is Presented

Context and goal: Poor wound healing is commonly associated with estrogen depletion during menopause. Experiments on anti-aging cosmetic formulations using genistein have yielded intriguing findings about skin health. Here, we examined the effects of systemically administered the genistein aglycones in an incisional wound healing model in comparison to systemically administered estradiol and raloxifene. Method of experimentation: Rats were randomly divided into groups of 12 animals each six months after Ovariectomies (OVX) and given daily treatments of raloxifene hydrochloride (0.05 and 0.5 mg·kg-1s.c.), genistein aglycone (1 and 10 mg·kg-1s.c.) or 17-a-ethinyl estradiol (0.003 and 0.03 mg·kg-1s.c.) for a period of 12 weeks. Rats with OVX and sham OVX were not treated and served as controls. Then, an incisional wound healing technique was carried out 14 or 7 days before to the experiment’s conclusion and skin specimens were gathered to assess molecular, histological and functional measurements. Important Results: Compared to samples from sham OVX animals, samples from OVX rats seven and fourteen days after wounding shown a decrease in transforming growthfactor-b1, tissue transglutaminase 2 and vascular endothelial growth factor. Genistein, raloxifene and estradiol all considerably altered this decline, but the lowest dose of genistein had a stronger impact than the other two therapies. Furthermore, the best genistein dosage for enhancing wound tensile strength and skin healing was the lowest one. Inferences and conclusions: One potential alternative treatment for the control of skin wound healing is genistein aglycone.

Effect of Genistein on Proliferating Cell Nuclear Antigen (PCNA) Expression and Vascular Density in The Peritoneum of Endometriosis Mice Model

Introduction: Survival of endometriosis lesions depends on the formation of blood vessels, cell proliferation, and oxygen supply. Their vascular densities can evaluate the vascularity of ectopic endometriosis tissues. Proliferating Cell Nuclear Antigen (PCNA) is one of the markers of cell proliferation that affects the growth of endometriotic lesions. Genistein is an isoflavone component that highly potential effect as an anti-angiogenic and inhibiting proliferation through the binding capacity to estrogen receptors. Material and Methods: This experimental study uses the Stored Biological Materials of mice peritoneum tissue. The paraffin block was divided into six groups, which are the negative control group, positive control group, and four treatment groups with genistein doses of 1.3 mg/day, 1.95 mg/day, 2.6 mg/day, and 3.25 mg/day. Immunohistochemical techniques and hematoxylin-eosin for vascular densities examined PCNA expressions. Preparation analysis uses OLIVIA software. Preparation calculations are done manually with cell count software. Observations were analyzed with the ANOVA test and Dunnett T3 5% test. Results: Based on analysis with ANOVA, there was a significant difference in PCNA expression and vascular densities because of the effect of genistein in various doses with a p-value of 0.001 (p<0.05). Based on Dunnett T3 5% test increase of PCNA expression by giving genistein doses 1,3 mg/day, 1,95 mg/day, and 2,6 mg/day, a reduction in dose of 3,25 mg/day. Vascular densities reduced significantly in all treatment groups. Conclusion: Administration of genistein in various doses significantly increases PCNA expression but reduces PCNA expression in a dose of 3.25 mg/day and significantly reduces vascular densities.

Genistein alleviates chronic heat stress-induced lipid metabolism disorder and mitochondrial energetic dysfunction by activating the GPR30-AMPK-PGC-1α signaling pathways in the livers of broiler chickens

The objective of this study was to investigate the preventive effects and mechanisms of genistein (GEN) on production performance and metabolic disorders in broilers under chronic heat stress (HS). A total of 120 male three-week-old Ross broilers were randomly assigned to 5 groups: a thermoneutral zone (TN) group maintained at normal temperature (21 ± 1°C daily), an HS group subjected to cyclic high temperature (32 ± 1°C for 8 h daily), and three groups exposed to HS with varying doses of GEN (50, 100, or 150 mg/kg diet). The experimental period lasted for three weeks. Here, HS led to a decline in growth performance parameters and hormone secretion disorders (P < 0.05), which were improved by 100 and 150 mg/kg GEN treatment (P < 0.05). Moreover, the HS-induced increases in the liver index (P < 0.01) and abdominal fat rate (P < 0.05) were attenuated by 150 mg/kg GEN (P < 0.05). The HS-induced excessive lipid accumulation in the liver and serum (P < 0.01) was ameliorated after 100 and 150 mg/kg GEN treatment (P < 0.05). Furthermore, the HS-induced decreases in lipolysis-related mRNA levels and increases in lipid synthesis-related mRNA levels in the liver (P < 0.01) were effectively blunted after 100 and 150 mg/kg GEN treatment (P < 0.05). Importantly, the HS-stimulated hepatic mitochondrial energetic dysfunction and decreases in the mRNA or protein levels of peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC-1α), nuclear respiratory factor 1, and mitochondrial transcription factor A in the liver were ameliorated by 150 mg/kg GEN (P < 0.05). Moreover, 50-150 mg/kg GEN treatment resulted in a significant increase in the mRNA or protein levels of G protein-coupled estrogen receptor (GPR30), AMP-activated protein kinase (AMPK) α1, phosphorylated AMPKα, and phosphorylated acetyl-CoA carboxylase α. Collectively, GEN alleviated metabolic disorders and hepatic mitochondrial energetic dysfunction under HS, possibly through the activation of GPR30-AMPM-PGC-1α pathways. These data provide a sufficient basis for GEN as an additive to alleviate HS in broilers.

Application of a skin and liver Chip2 microphysiological model to investigate the route-dependent toxicokinetics and toxicodynamics of consumer-relevant doses of genistein.

The HUMMIC skin-liver Chip2 microphysiological system using EpiDerm™ and HepaRG and stellate liver spheroids was used to evaluate the route-specific metabolism and toxicodynamic effects of genistein. Human-relevant exposure levels were compared: 60 nM representing the plasma concentration expected after topical application of a cosmetic product and 1μM representing measured plasma concentrations after ingesting soya products. Genistein was applied as single and repeated topical and/or systemic doses. The kinetics of genistein and its metabolites were measured over 5days. Toxicodynamic effects were measured using transcriptional analyses of skin and liver organoids harvested on Days 2 and 5. Route-specific differences in genistein's bioavailability were observed, with first-pass metabolism (sulfation) occurring in the skin after topical application. Only repeated application of 1μM, resembling daily oral intake of soya products, induced statistically significant changes in gene expression in liver organoids only. This was concomitant with a much higher systemic concentration of genistein which was not reached in any other dosing scenario. This suggests that single or low doses of genistein are rapidly metabolised which limits its toxicodynamic effects on the liver and skin. Therefore, by facilitating longer and/or repeated applications, the Chip2 can support safety assessments by linking relevant gene modulation with systemically available parent or metabolite(s). The rate of metabolism was in accordance with the short half-life observed in in vivo in humans, thus supporting the relevance of the findings. In conclusion, the skin-liver Chip2 provides route-specific information on metabolic fate and toxicodynamics that may be relevant to safety assessment.

Effect of High-intensity Interval Training and Genistein on Antioxidant Capacity in the Heart Tissue of Elderly Rats

Background: Aging can lead to a decrease in cardiac function and an increase in oxidative stress in the heart, which leads to cellular aging through mitochondrial dysfunction. The present study aimed to investigate the antioxidant effects of high-intensity interval training (HIIT) and genistein (Ge) consumption in the heart tissue of elderly rats. Materials and Methods: In this experimental study, 30 female Sprague-Dawley rats aged 18 to 24 months were selected and divided into five groups of six rats, including 1) HIIT, 2) Ge, 3) HIIT + Ge, 4) control, and 5) sham. During eight weeks, groups 2 and 3 received 60 mg of Ge dissolved in DMSO per kilogram of body weight daily, and group 5 received 60 mg of DMSO per kilogram of body weight per day intra-peritoneally. In addition, groups 1 and 3 ran on the treadmill three times per week. For statistical analysis of data, Shapiro-Wilk, independent samples t-test, one-way ANOVA, and Tukey's post-hoc test were used. Results: High-intensity interval training had a significant effect on increasing GPx and MDA (P&lt;0.05), Ge led to a significant decrease in SOD (P&lt;0.05); however, HIIT with Ge had not interactive effects on improving SOD, GPx, and MDA (P&lt;0.05). Conclusion: It seems that the antioxidant system is dependent on the intensity of exercise and the dose of genistein. Therefore, HIIT and Ge in this study have no interactive effect on the oxidant-antioxidant system in the heart of elderly rats.

Genistein Effect on Estrogen Receptor especially Interleukin-6 and Tumor Necrosis Factor-α in Mice Model of Endometriosis: a Systematic Review and Meta-analysis

Introduction: Endometriosis is the endometrial tissue proliferation outside the uterine space. Anti-angiogenic agents from different groups of substances are currently being discussed as promising therapies for the treatment of endometriosis in the future, one of which is therapy using the phytoestrogen genistein. At the molecular level, genistein's effects on cells include inhibition of proliferation, induction of differentiation, apoptosis, and cell arrest in the cell cycle, which are related to the transforming growth factor. Genistein inhibits cell growth by modulating the signal-transforming growth factor pathway. Methods: We searched PubMed, Scopus, Cochrane Library and Proquest from its inception to December 2022 to collect observational and interventional studies of Genistein consumption in endometriosis mice. Two authors independently screened the literature according to the inclusion and exclusion criteria, extracted data, and assessed the risk of bias. Safety analysis was assessed through TNF-α and IL-6 values of estrogen expression. Results: No significant difference in the values of IL-6 and TNF-α in rats with endometriosis that were given genistein at these three doses. However, if analyzed further, the IL-6 outcome compared to the 0.78mg/day dose most affected the IL-6 value between the two groups, and the TNF-α output compared to the 0.78 mg/day dose most affected the TNF-α value between the two groups. So, a genistein dose of 0.78 mg/day can affect IL-6 and TNF-α in mice with endometriosis. Conclusion: Genistein modulated estrogen expression of TNF-α and IL-6 at all doses, but the best dose for its effect was 0.78 mg/day.

Rottlerin and genistein inhibit neuroblastoma cell proliferation and invasion through EF2K suppression and related protein pathways.

Neuroblastoma is one of the most common solid tumors in children younger than 1year of age, with poor prognosis and survival rates. Therefore, novel molecular targets and therapeutic strategies are needed to prolong patient survival. For this purpose, we investigated the effects of rottlerin and genistein separately and in combination on neuroblastoma cells (SH-SY5Y, Kelly). First, the effects of rottlerin and genistein were investigated on cell proliferation. Different rottlerin (1-50µM) and genistein (5-150µM) doses were used as experimental groups compared to the control (DMSO/vehicle). The IC50 dose was found to be 5µM for rottlerin and 30µM for genistein (P < 0.0001). Other analyses, such as colony formation assays, annexin V/propidium iodide staining, matrigel invasion assays, and Western blot analysis, were performed with these doses and their combinations. To assess statistical significance, statistical analysis was conducted using the one-way ANOVA with the post hoc Tukey test. Our results showed that IC50 doses of rottlerin and genistein induced a significant reduction in cell proliferation, colony formation, and invasion in neuroblastoma cells (P < 0.0001). The combination of these doses increased the levels of inhibition of cell proliferation and invasion while decreasing the level of apoptosis (P 0.0001). Furthermore, these agents caused G1-cell cycle arrest in these cells. Our western blot data showed that rottlerin and genistein treatments markedly inhibit elongation factor 2 kinase (EF2K) and other pro-tumorigenic, metastatic proteins in neuroblastoma cells. These agents probably showed their anti-proliferative, anti-metastatic, and pro-apoptotic effects through EF2K downregulation. Our results suggested that rottlerin and genistein have inhibitory effects on cancer cell proliferation, invasion, and cell cycle and induce apoptosis in both cell lines. Combined treatment with rottlerin and genistein may be a viable approach and beneficial to neuroblastoma patients as the combined effect significantly suppresses the above-mentioned pathways.

Genistein early in life modifies the arcuate nucleus of the hypothalamus morphology differentially in male and female rats

In the present work we analyzed the effects of postnatal exposure to two doses of genistein (10 μg/g or 50 μg/g) from postnatal (P) day 6 to P13, on the morphology of the arcuate nucleus (Arc). The analyses of Arc coronal brain sections at 90 days showed that the ArcMP had higher values in volume, Nissl-stained neurons and GPER-ir neurons in males than in females and the treatment with genistein abolished these sex differences in most of the parameters studied. Moreover, in males, but not in females, the GPER-ir neurons decreased in the ArcMP but increased in the ArcL with both doses of genistein. In the ArcLP, GPER-ir population increased with the lowest doses and decreased with the highest one in males. Our results confirm that the Arc subdivisions have differential vulnerability to the effects of genistein during development, depending on which neuromorphological parameters, dose and sex are analyzed.

Effects of genistein supplemented before or after irradiation on DNA injury in human lymphocytes in vitro.

Ionizing radiation (IR) carry adequate energy to ionize or remove electrons from an atom. Particles interact with water to produce reactive oxygen species (ROS). Genistein (GEN) is a naturally occurring phytoestrogen and the basic isoflavonoid in soybeans and soybean-enriched products and is believed to have the strongest antioxidant activity. The study aimed at the investigation if application of GEN at different time prior or past irradiation may ameliorate or reduce injury of DNA in human lymphocytes. The isolated lymphocytes were exposed to X-irradiation (0.5; 1 Gy). GEN (1 µM/ml; 10 µM/ ml) was appended to attempts at various times prior or past irradiation (1 h prior, immediately prior, immediately past, 1 h past). We joined each X-rays dose with each GEN dose. After 1h of incubation DNA damages were examined using Comet assay. Combination of 1 µM/ml of GEN given 1 h before irradiation with low or high dose markedly decreased induced by irradiation DNA injury. Higher dose of GEN applied immediately before or after irradiation markedly extended the frequency of DNA injury generated by irradiation. The result of application 1 µM/ml GEN 1 h after irradiation was not significantly different compared to control. The effect of 1 Gy + 10 µM/ml GEN was not significantly lower compared to each agent alone. Only a very low concentration of GEN applied before irradiation, may be considered as a potential radiomitigator/radioprotector. High doses of GEN work as a radiosentitizer and may potent the effects of radiotherapy.

Genistein administration increases the level of superoxide dismutase and glutathione peroxidase in the endometriosis mice model: An experimental study.

Endometriosis and infertility are caused by reactive oxygen species or free radicals, which promote endometrial cell growth and adhesion in the peritoneal cavity. Genistein has been proven to protect cells against reactive oxygen species by scavenging free radicals and decreasing the expression of genes-associated stress responses. This study was conducted to determine whether genistein also acts as an antioxidant by elevating superoxide dismutase (SOD) and glutathione peroxidase (GPx) in the peritoneal fluid of the endometriosis mice model. This experimental study involved 32 healthy female mice (Mus musculus), aged between 2-3 months and weighing 20-30 gr. They were divided into negative control group (healthy mice without genistein), endometriosis group (endometriosis mice without genistein), treatment group that was given different doses of genistein, that is, 0.13; 0.26; 0.52; 0.78; 1.04; and 1.3 mg/day (n = 4/each). SOD level in the peritoneal fluid was measured using the quantitative colorimetric determination method, and a colorimetric assay measured the GPx levels. Results showed that the endometriosis model has lower SOD and GPx levels than the control group. The administration of genistein significantly normalized these changes. Genistein significantly increased SOD levels in the 0.13 mg and 0.26 mg treatment groups. Genistein also increased GPx levels significantly in all treatment groups. Genistein increases SOD and GPx levels in the peritoneal fluid of an endometriosis mice model, and the change is dose-dependent.

10-7M genistein partially alleviates 10-7M MEHP unfavorable effects in a new modified fetal rat testis culture system.

Background: Recent studies revealed that some common endocrine-disrupting chemicals (EDCs) including phthalates and phytoestrogens may exhibit low-dose effects properties. However, how low dose of these EDCs and their mixture would affect fetal rat testis development still needs further investigation. Moreover, testis organ culture system also needs further modification to provide an effective tool for ex vivo EDCs study. Methods: We firstly modified the agarose organ culture system, in which fetal rat testes were cultured for 4 days (d1 to d4) on agarose gels held by Millicell inserts. Then we used the modified agarose culture system to study the combined effects of multiple EDCs exposure. 15.5 dpc fetal rat testes were isolated and treated with vehicle, MEHP (0.1 μmol/L), GEN (0.1 μmol/L) or MEHP (0.1 μmol/L) + GEN (0.1 μmol/L). Parameters concerning testicular cell development and function were evaluated, trying to gain insight into the early molecular events after multiple EDCs exposure. Results: The development of somatic, germ cells and seminiferous tubule in 15.5 dpc fetal rat testis was better sustained in the modified agarose culture system. Based on the modified system, we found that MEHP at 0.1 μmol/L induced alterations in gonocyte markers, antioxidative enzyme activity as well as transient reduction of testosterone production, accompanied by mitochondria swelling in gonocytes and Sertoli cells. No obvious morphological and histological alterations were observed in all treated groups. However, coadministration of genistein at 0.1 μmol/L partially alleviated MEHP-induced fetal testis damage ex vivo through enhancement of antioxidative action. MEHP at low dose still showed weak endocrine disrupting properties but did not exhibit typical low-dose effects. Conclusion: Our findings indicated that the modified agarose culture system could better mimic testicular microenvironment without obvious hypoxic cell damage. Furthermore, low dose of MEHP induced mild disruption to fetal testis development, cotreatment of genistein at low dose attenuated MEHP induced fetal testis injuries in part by balancing redox state, indicating that low dose of genistein may partially protect fetal testis from phthalates induced injury.

Phloretin-induced suppression of oxidative and nitrosative stress attenuates doxorubicin-induced cardiotoxicity in rats

Objective: To compare the cardioprotective efficacy of equimolar doses (50 mM/kg, p.o.) of phloretin and genistein against doxorubicin-induced cardiotoxicity in rats. Methods: Cardiotoxicity was induced in rats by intraperitoneal injection of 6 mg/kg doxorubicin on alternative days till the cumulative dose reached 30 mg/kg. This study included four treatment groups of rats (n=6): the control group (0.5% carboxymethyl cellulose solution-treated), the doxorubicin- treated group (0.5% carboxymethyl cellulose solution along with doxorubicin), the genistein-treated group (50 mM/kg/day; p.o. along with doxorubicin) and phloretin-treated group (50 mM/kg/day; p.o. along with doxorubicin). On the 10th day of dosing, rats were anesthetized for recording ECG, mean arterial pressure, and left ventricular function. Oxidative stress, nitric oxide levels, and inflammatory cytokines were estimated in the cardiac tissue. Cardiac function parameters (creatine kinase MB, lactate dehydrogenase, aspartate aminotransferase, and alanine transaminase) were estimated in the serum samples. Results: Phloretin treatment inhibited doxorubicin-induced oxidative stress and also reduced nitric oxide levels in cardiac tissues of rats. Phloretin administration attenuated doxorubicin- induced alterations in hemodynamic parameters (heart rate, mean arterial blood pressure, and left ventricular function) and suppressed the expression of pro-inflammatory cytokines. The cardiac injury markers like creatine kinase MB, lactate dehydrogenase, aspartate aminotransferase, and alanine transaminase were reduced by both genistein and phloretin. All these effects of phloretin were more prominent than genistein. Conclusions: Phloretin offers cardioprotection that is comparable to genistein, a clinically validated cardioprotectant against doxorubicin-induced cardiotoxicity. Further studies are needed to confirm and establish the therapeutic utility of phloretin as a chemopreventive adjuvant to doxorubicin chemotherapy.

Effects of genistein on glucose and lipid metabolism of common carp (Cyprinus carpio. L) in vivo and in vitro

Genistein exhibits anti-diabetic and anti-lipogenic effects in mammals, however, there are rare reports on the effects of genistein on glucose and lipid metabolism in fish. This study explored the regulation role of genistein on glucose and lipid metabolism of common carp. After oral gavage with genistein (20 mg/kg body weight), the content of genistein in serum was the highest at 4 h and rapidly returned to basal values at 48 h. The blood glucose (GLU) concentrations were decreased at 24 h after gavage with genistein. The total cholesterol (TC) level was significantly down-regulated by genistein except for 24 h (P < 0.01). On the contrary, serum triglycerides (TG), LDL-C, and HDL-C were significantly up-regulated after treatment (P < 0.05). The mRNA level of the genes related to gluconeogenesis, glycogen synthesis, and lipolysis was significantly inhibited after treatment with genistein. Meanwhile, genistein administration has up-regulated the level of lipid synthesis-related genes. To compare the effects of genistein with estradiol, the primary hepatocytes were incubated with different concentrations of genistein (1, 10, or 100 µM) and 17β-estradiol (E2, 1 µM) respectively. Similar to the in vivo results, the lipid synthesis-related genes levels were increased and lipolysis genes were decreased after treatment with a high dose of genistein (100 µM). On the contrary, the genes involved in lipid synthesis were down-regulated, while lipolysis genes were up-regulated at the low and medium doses of genistein (1 and 10 µM). Moreover, low and medium doses of genistein enhanced the expression of gluconeogenesis-related genes. There is a certain similar effect of estrogen and genistein in vitro, it is speculated that genistein may play an estrogen-like role in regulating glucose and lipid metabolism. The effect of genistein on fish glucose and lipid metabolism may be affected by the concentration level, treatment time, or the methods.

Genistein Regulates Lipid Metabolism via Estrogen Receptor β and Its Downstream Signal Akt/mTOR in HepG2 Cells.

Genistein (GEN) has been shown to significantly inhibit hepatic triglyceride accretion triggered by estrogen deficiency. The main purpose of this in vitro study was to investigate the function and molecular mechanism of estrogen receptor β (ERβ) in regulating hepatic lipid metabolism induced by GEN. Different doses of GEN or GEN with an ERβ antagonist were treated with HepG2 cells. Results showed that 25 μM GEN significantly diminished triglyceride levels. Meanwhile, GEN downregulated the levels of genes and proteins involved in lipogenesis, such as sterol-regulatory element-binding protein-1c (SREBP-1c), fatty acid synthase (FASN), and stearoyl-coenzyme A desaturase 1 (SCD1), and upregulated the gene and protein levels of the regulation factors responsible for fatty acid β-oxidation, such as carnitine palmitoyltransferase 1α (CPT-1α) and peroxisome proliferator-activated receptor α (PPARα). Furthermore, 25 μM GEN reduced the levels of phosphorylation of protein kinase B (Akt) and mechanistic target of rapamycin (mTOR). Moreover, most of these effects from GEN were reverted by pretreatment with the antagonist of ERβ. In conclusion, GEN improved hepatic lipid metabolism by activating ERβ and further modulation of Akt/mTOR signals. The results provide novel aspects of the regulatory mechanism of ERβ on hepatic lipid metabolism and might help to profoundly understand the functions of food-derived phytoestrogens in preventing and treating hepatic steatosis in postmenopausal women.

The Effect of Soya and Genistein Supplementation on Serum Lipid Profile of Male Dawley Sprague Rat

Background: Non-medical therapy is needed to reduce blood lipid levels and prevent atherosclerosis. Foods that contain Isoflavones can improve lipid profiles in the blood.Aim: To determine the effect of soya and genistein supplementation in various doses on total cholesterol, Low Density Lipoprotein (LDL), High Density Lipoprotein (HDL), Triglycerides in male Sprague-Dawley rats.Methods: 35 male Sprague-Dawley rats aged 6-8 weeks were divided into 7 groups, control group; low, medium, and high doses of soy milk; and low, moderate, and high doses of genistein. The treatment was given orally for 60 days. The control used was mice with standard feed. On the 61st day, blood and hepatic tissue samples were taken and checked for lipid profiles using the Friedewald formula. Statistical analysis was carried out by one-way ANOVA and Tukey's posthoc test and Dunnet test, then regression analysis.Results: There was a significant increase in total cholesterol levels (p &lt;0.05) and  LDL levels in low and moderate doses of soy milk group compared to all groups of genistein. There was no significant differences in HDL levels in soya and genistein groups. There was significant increase in Triglyceride levels in all groups of soy milk and low and moderate doses of Genistein compared to high doses of genistein.Conclusion:  Soya and genistein supplementation had increased effect on the Total Cholesterol, LDL and Triglycerides but no effect on HDL in all groups.

Estrogenic and vitellogenic responses in genistein fed adult male Cyprinus carpio

A 60-day feeding trial was conducted to determine the impact of dietary phytoestrogen, genistein on inducing vitellogenesis in reproductively active male Cyprinus carpio. Adult male C. carpio were fed with graded levels of genistein, 0 (Control), 1 (Gen-1), 3 (Gen-3), 6 (Gen-6) and 9 (Gen-9) mg/100 g in fish feed. Serum levels of cholesterol, total testosterone and estradiol (E2) showed an increasing trend with increasing doses of genistein in the feed after the trial. Serum T3 level was reduced while the T4 level was induced with an increasing level of dietary genistein. Gene expression of brain and testicular aromatases were significantly reduced in Gen-9. The expression of estrogen receptor-α (erα) was upregulated, and that of erβ2 was downregulated with an increasing dose of genistein. Vitellogenin concentration was increased to its highest, 29.70 ± 2.63 μg/mL in Gen-6 and vtgb2 expression in the liver was increased to five-fold in Gen-6 & Gen-9 than that of control. The level of cortisol and the expression pattern of 20βHSD were reduced with an increasing dose of genistein. Hence, the study concluded that upregulation of the erα gene and high E2 concentration is responsible for genistein induced vitellogenesis in adult male C. carpio. The estrogenic responses in adult male C. carpio are accompanied by enhanced testosterone level and reduced activity of the steroidogenic enzyme, 20βHSD, indicating a possible delay in maturation.