Addition Of Genistein Research Articles

Adding genistein or luteolin decreased the yield of citrinin and without reducing pigments in yam solid-fermentation by Monascus.

Chinese yam fermented by Monascus, namely red mold dioscorea (RMD), has the potential of treating diseases. However, the production of citrinin limits the application of RMD. In the present study, the fermentation process of Monascus was optimized by adding genistein or luteolin to reduce citrinin yield. The results showed that citrinin in 25 g of Huai Shan yam was reduced by 48% and 72% without affecting the pigment yield by adding 0.2 g of luteolin or genistein, respectively, to a 250-mL conical flask after fermentation for 18 days at 28 °C, whereas the addition of luteolin increased the content of yellow pigment by 1.3-fold. Under optimal conditions, citrinin in 20 g of iron bar yam decreased by 55% and 74% after adding 0.2 g of luteolin or genistein. Luteolin also increased yellow pigment content by 1.2-fold. Ultra HPLC coupled to quadrupole time-of-flight mass spectrometry was used for the preliminary analysis of Monascus fermentation products. It was found that the amino acid types in RMD are similar to those in yams, but there are fewer polysaccharides and fatty acids. The results obtained in the present study showed that the addition of genistein or luteolin could reduce citrinin on the premise of increasing pigment yield, which laid a foundation for the better use of yams in Monascus fermentation. © 2023 Society of Chemical Industry.

Enhancement of functional activity and biosynthesis of exopolysaccharides in Monascus purpureus by genistein treatments

The exopolysaccharides (EPS) produced by the edible medicinal fungus Monascus purpureus (EMP) become the center of growing interest due to their techno-functional properties and their numerous applications in the food industries; however, the low EPS yields limit its application. In this study, the effect of genistein supplementation on the production, rheological and antioxidant properties of EPS by M. purpureus and its biosynthesis mechanism were explored. The results indicated that the addition of genistein (3 g/L) generated a 110% and 59% increase in the maximum mycelial biomass and EPS yield, respectively. The genistein supplementation group (G-EMP) had higher molar percentages of Xyl and Man, and significantly decreased molecule weight and particle size of EPS, which resulted in stronger antioxidant effect and cell growth promotion. Rheological analysis showed that both EMP and G-EMP demonstrated pseudoplastic fluid behavior and G-EMP exhibited strong gel-like elastic behavior (G' > G"). Furthermore, genistein not only facilitated the production of EPS by regulating cell membrane permeability, enhancing cellular respiratory metabolism and monosaccharide precursor synthesis pathways, and enhancing antioxidant enzyme activity to reduce oxidative stress damage, but also affected the composition of the monosaccharides by increasing enzyme activity in the underlying synthesis pathways. These findings expand the application of M. purpureus resources and provide a paradigm for future study of the structural and functional characteristics of EPS.

Role of genistein on the yield, structure and immunomodulatory activity of Monascus exopolysaccharides.

Manipulating the structures, physicochemical properties, and monosaccharide compositions of exopolysaccharides (EPS) isolated from microorganisms has been reported to enhance their biological activities. Hence, the aim of this work was to examine the effects of genistein addition during fermentation on the amount, physicochemical properties, and immunomodulatory activity of EPS secreted by M. purpureus. Results showed that genistein addition significantly increased M. purpureus biomass and EPS yield to 2.42 g L-1 and 6.08 g L-1, respectively, and affected the physicochemical properties and structures of EPS. Furthermore, EPS produced by genistein-treated M. purpureus (G-EMP) improved the immunomodulatory activity of RAW264.7 macrophages by increasing the secretion of nitric oxide and cytokines. Moreover, phospho-Jun N-terminal kinase (p-JNK), phospho-extracellular regulated protein kinase (p-ERK), phospho-p38 (p-p38) mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) phospho-p65 (p65) proteins were remarkably upregulated by G-EMP stimulation, blocking Toll-like receptor 4 (TLR4) that dramatically reduced the pinocytic and phagocytic capacities. Overall, these findings provide potential rationales for the application of genistein in improving the EPS yield of M. purpureus.

Genistein inhibits the proliferation of human choriocarcinoma cells via the downregulation of estrogen receptor-α phosphorylation at serine 118

<h2>Summary</h2><h3>Background & aims</h3> Choriocarcinoma is a malignant trophoblastic tumor. The phosphorylation of estrogen receptor-α at serine 118 (p-ER-s118) decreases cancer cell proliferation. However, the effect of genistein as a modulator of p-ER-s118 and proliferation of chorioarcinoma cells remains to be understood. This study aims at determining the function of genistein on p-ER-s118 levels and human choriocarcinoma JEG-3 cell proliferation. <h3>Methods</h3> After reaching confluency, cells were divided into six groups, the control group (without methyl-piperidino-pyrazole (MPP) pre-treatment and genistein treatment); and groups with cells treated with genistein at concentrations of 0, 10, 25, 50, and 100 μM (cells were pretreated with MPP). Expression of p-ER-s118 and Ki-67 were analyzed using immunocytochemistry. <h3>Results</h3> Different doses of genistein decreased p-ER-s118 levels compared to those in the control (<i>p</i> < 0.05). JEG-3 cell proliferation was inhibited by MPP pre-treatment, concomitant with genistein treatment with a dose of 0 μM, 25 μM, 50 μM, and 100 μM compared to the proliferation of the control cells (<i>p</i> < 0.05). <h3>Conclusion</h3> Taken together, treatment with methyl-piperidino-pyrazole downregulated p-ER-s118. The addition of genistein further decreased the levels of p-ER-s118 and inhibited cell proliferation. Thus, <i>in vivo</i> studies nedd to follow this <i>in vitro</i> study to elucidate the mechanism(s) employed by genistein as an alternative therapy for choriocarcinoma.

Functional analysis of isoflavones using patient-derived human colonic organoids

Inflammatory bowel disease (IBD) comprises two major subtypes, ulcerative colitis (UC) and Crohn’s disease, which are multifactorial diseases that may develop due to genetic susceptibility, dysbiosis, or environmental factors. Environmental triggers of IBD include food-borne factors, and a previous nationwide survey in Japan identified pre-illness consumption of isoflavones as a risk factor for UC. However, the precise mechanisms involved in the detrimental effects of isoflavones on the intestinal mucosa remain unclear. The present study employed human colonic organoids (hCOs) to investigate the functional effect of two representative isoflavones, genistein and daidzein, on human colonic epithelial cells. The addition of genistein to organoid reformation assays significantly decreased the number and size of reformed hCOs compared with control and daidzein treatment, indicating an inhibitory effect of genistein on colonic cell/progenitor cell function. Evaluation of the phosphorylation status of 49 different receptor tyrosine kinases showed that genistein selectively inhibited phosphorylation of epidermal growth factor receptor (EGFR) and hepatocyte growth factor receptor (HGFR). We established a two-dimensional wound-repair model using hCOs and showed that genistein significantly delayed the overall wound-repair response. Our results collectively show that genistein may exert its detrimental effects on the intestinal mucosa via negative regulation of stem/progenitor cell function, possibly leading to sustained mucosal injury and the development of UC.

Addition of genistein to the fermentation process reduces citrinin production by Monascus via changes at the transcription level

Monascus, which is traditionally used in various Asian industries, produces several secondary metabolites during the fermentation process, including citrinin, a toxin whose impact limits the development of the Monascus industry. We have previously found that the addition of 2.0 g/L genistein to Monascus medium reduces citrinin production by approximately 80%. Here, we explored the molecular mechanisms whereby genistein affects citrinin production. We sequenced the Monascus genome and performed transcriptome analysis on genistein-treated and -untreated groups. Comparison between the two groups showed 378 downregulated and 564 upregulated genes. Among the latter, we further examined the genes related to citrinin biosynthesis and quantified them using quantitative real-time polymerase chain reaction (qRT-PCR). Genes orf5, pksCT, orf3, orf1, orf6, and ctnE were significantly downregulated, demonstrating that genistein addition indeed affects citrinin synthesis. Our results may lay the groundwork for substantial improvements in the Monascus fermentation industry.

Metabolomics analysis based on UHPLC-Q-TOF-MS/MS reveals effects of genistein on reducing mycotoxin citrinin production by Monascus aurantiacus Li AS3.4384

Monascus mold produces useful bioactive compounds such as Monascus pigments, monacolin K, and ergosterol. In this study, metabolomics analysis was used to investigate the metabolic profile changes in Monascus aurantiacus Li AS3.4384 (MALA) induced by genistein at different fermentation times using UHPLC-Q-TOF-MS/MS. The addition of genistein was found to not only reduce citrinin but also suppress significant differential metabolites. A total of 21 significant differential metabolites were involved in the most significant metabolic pathways (impact >0.1). These metabolites affected the aspartic acid, glutamic acid pathway, tricarboxylic acid (TCA) cycle, pyruvate synthesis pathway, and lysine degradation pathway. Thus, the addition of genistein may change the precursor for citrinin synthesis of Monascus. These findings provide significant insight into the Monascus metabolic pathways affected by genistein and will help to improve the quality control and safety of Monascus in food and supplements.

The Effect of Genistein on the Plasma Membrane Integrity of Frozen Ongole Grade Bull Semen Based on Skim Milk – Soy Lecithin Extender

Beef cattle represent livestock commodities that can help support national economic stability. Ongole grade bull (PO) is one of the local cattle breeds whose population and genetic quality decrease. Various efforts have been made in improving the genetic quality and in increasing the population of the local livestock population to achieve self-sufficient status of the beef supply in Indonesia. One of the efforts is artificial insemination (IB). The study aims at examining the effect of different concentrations of genistein semen extender on the post-thawing membrane integrity of Ongole grade bull semen. It is useful in developing the genistein as a protective agent against reactive oxygen species (ROS) during cryopreservation process. It was conducted at BPBPTDK, Center for Animal Breeding Development, Animal Feed, and Veterinary Diagnostics, in the Special Region of Yogyakarta from February 7th, 2019 to May 7th, 2019. The semen of two healthy Ongole grade bulls was consecutively collected five times in the period of 3 months using artificial vagina. The semen was cryopreserved using skim milk-soy lecithin semen based extenders at different concentrations of genistein, which were group A (1.0 mmol genistein), B (2.0 mmol genistein) and K (without any genistein as control). The observed parameters included individual spermatozoa motility, percentage of spermatozoa viability that was observed using Eosin nigrosine staining method and spermatozoa membrane integrity that was observed using the hypo osmotic swelling test (HOST) method. The observations were made on fresh semen, extended semen, and post-thawing semen. The results of the observation were then analyzed descriptively and statistically using regression correlation test and one-way analysis of variance (ANOVA). The results of the analysis showed that the addition of Genistein at the different concentrations of 1 mmol and 2 mmol resulted in higher motility, viability, and membrane integrity than the control and had significant effect (p<0.05) on the motility, the membrane integrity, and the acrosome integrity of the Ongole grade bull semen. The genistein concentration of 2.0 mmol gave the highest mean percentage of spermatozoa motility (57.50 ± 1.00 %) after freezing. The genistein concentration of 1.0 mmol gave the highest mean percentage of spermatozoa membrane integrity and acrosome integrity (52.45 ± 0.85 % and 49.58 ± 0.98 %) after freezing.

A Possible Mechanism: Genistein Improves Metabolism and Induces White Fat Browning Through Modulating Hypothalamic Expression of Ucn3, Depp, and Stc1.

Bioactive food components have gained growing attention in recent years. Multiple studies demonstrated that genistein had beneficial effects on metabolism. However, the exact mechanism by which genistein improves metabolism remains unclear, especially the central regulation. This study was designed to evaluate whether addition of genistein to the high-fat diet could counter metabolic disorders and whether these alterations were associated with gene expression in hypothalamus. C57BL/6 mice were fed either a high-fat diet (HF), high-fat diet with genistein (0.25 g/kg diet) (HFG) or a normal control diet (CON) for 8 weeks. Body weight was assessed during the study. After 8-week intervention, content of inguinal subcutaneous adipose tissue (SAT), perirenal visceral adipose tissue (VAT) and brown adipose tissue (BAT) were weighed. Glucose tolerance test, the serum levels of insulin and lipid were assessed. The mRNA of browning marker was detected in the white fat. The hypothalamus was collected for whole transcriptome sequencing and reverse transcription quantitative PCR validation. The results demonstrated that mice fed HFG diet had lower body weight and SAT mass, decrease levels of low-density lipoprotein cholesterol and free fatty acids, higher browning marker of Ucp1 and Cidea in WAT and an improvement in glucose tolerance and insulin sensitivity compared with those in HF group. Transcriptome sequencing showed that there were three differentially expressed genes in hypothalamus among the three groups, including Ucn3, Depp, and Stc1, which were significantly correlated with the browning markers in WAT and insulin sensitivity. Thus, regulating gene expressions in hypothalamus is a potential mechanism for genistein improving metabolism and inducing WAT browning, which may provide a novel target for the precaution and treatment of T2DM.

1821-P: A Possible Mechanism: Genistein Improves Metabolism and Induces White Fat Browning through Modulating Hypothalamic Expression of Ucn3, Depp, and Stc1

Bioactive food components have gained growing attention in recent years. Multitude of studies have demonstrated that genistein, as a soy-derived phytoestrogen, has beneficial effects on metabolism. However, the exact mechanism by which genistein improving metabolic disorders remains unclear, especially the central regulation. This study was designed to evaluate whether addition of genistein to the high-fat diet could counter the metabolic disorders and whether these alterations were associated with the gene expression in hypothalamus. C57BL/6 mice were fed either a high-fat diet (HF), high-fat diet with genistein (0.25g/kg diet) (HFG) or a normal control diet (CON) for eight weeks. Body weight were assessed during the study. After eight-week intervention, content of subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT) and brown adipose tissue (BAT) were weighed. Glucose tolerance test, the serum levels of insulin and lipid were assessed. The mRNA of browning marker was detected in the WAT. The hypothalamus was collected for whole transcriptome sequencing and reverse transcription quantitative PCR (RT-qPCR) validation. The results demonstrated that mice fed the HFG diet had lower body weight (p&amp;lt;0.05) and SAT mass (p&amp;lt;0.01), decrease levels of low-density lipoprotein cholesterol (LDL-C) (p&amp;lt;0.01) and free fatty acids (FFA) (p&amp;lt;0.05), higher browning marker of Ucp1 (p&amp;lt;0.01) and Cidea (p&amp;lt;0.05) in WAT and an improvement in glucose tolerance and insulin sensitivity compared with those in the HF group. Transcriptome sequencing showed that there were three differentially expressed genes in hypothalamus among the three groups, including Ucn3 (p&amp;lt;0.05), Depp (p&amp;lt;0.01) and Stc1 (p&amp;lt;0.01). Thus, regulating gene expressions in hypothalamus is a potential mechanism for genistien improving metabolism and inducing WAT browning, which may provide a novel target for the precaution and treatment of T2DM. Disclosure L. Zhou: None. X. Xiao: None. M. Deng: None. J. Liu: None. M. Yu: None. Funding National Natural Science Foundation of China

Effect of genistein on IGF‐1 and IGFBP‐1 in young and aged female rat ovary

The objective of this study was to assess the effects of genistein (GEN) on expression of insulin-like growth factor 1 (IGF-1) and insulin-like growth factor binding protein 1 (IGFBP-1) in young and aged rat ovary. Forty young female Sprague Dawley (SD) rats (200±20g) and forty aged female SD rats (490±20g) were selected and according to weight, they were divided into the following five groups with eight animals in each: negative control group (NC), low-dose group (L), middle-dose group (M), high-dose group (H) and positive control group (PC). GEN group received GEN of 15, 30, 60mg/kg respectively. It lasted 30days. Concentrations of serum hormones, IGF-1 and IGFBP-1 were determined by enzyme-linked immunosorbent assay (ELISA). Gene and protein expressions of IGF-1 and IGFBP-1 were determined by real-time PCR and Western blot respectively. Compared with NC, GEN significantly increased oestradiol-17β(E2 ) level in aged rat, reduced luteinizing hormone (LH) level in young and aged rat. Serum levels of IGFBP-1 in young rats were significantly higher in GEN groups (p<0.05). mRNA and protein expression levels of IGF-1 and IGFBP-1 were positively correlated with GEN dose. GEN could significantly reduce the ratio of IGF-1/IGFBP-1 of aged rats. Multivariate Cox regression analysis result showed IGF-1 and IGFBP-1 levels significantly correlated with GEN dose. We speculate that there is an association between the addition of GEN and expression of IGF-1 and IGFBP-1, and the relationship between them is different in young and aged rat.

Malaria pigment accelerates MTT - formazan exocytosis in human endothelial cells.

Haemozoin is a by-product of haemoglobin digestion by intraerythrocytic malaria parasites, which induces immunologic responses on different tissues, including endothelial cells. In the present paper, the incubation of human microvascular endothelial cells with haemozoin significantly inhibited MTT reduction, a measure of cytotoxicity, without increasing the release of cytoplasmic lactate dehydrogenase. Moreover, haemozoin did not induce apoptosis or cell cycle arrest nor decreased the number of live cells, suggesting that cells viability itself was not affected and that the inhibition of MTT reduction was only apparent and probably due to accelerated MTT-formazan exocytosis. After 30 min of MTT addition, a significant increase in the % of cells exocytosing MTT formazan crystals was observed in haemozoin-treated cells compared with control cells. Such an effect was partially reversed by the addition of genistein, an inhibitor of MTT-formazan exocytosis. The rapid release of CXCL-8, a preformed chemokine contained in Weibel-Palade bodies, confirmed that haemozoin induces a perturbation of the intracellular endothelial trafficking, including the exocytosis of MTT-formazan containing vesicles. The haem moiety of haemozoin is responsible for the observed effect. Moreover, this work underlines that MTT assay should not be used to measure cytotoxicity induced by haemozoin and other methods should be preferred.

Long-Term Genistein Consumption Modifies Gut Microbiota, Improving Glucose Metabolism, Metabolic Endotoxemia, and Cognitive Function in Mice Fed a High-Fat Diet.

The aim of this study is to assess whether the long-term addition of genistein to a high-fat diet can ameliorate the metabolic and the cognitive alterations and whether the changes can be associated with modifications to the gut microbiota. C57/BL6 mice were fed either a control (C) diet, a high-fat (HF) diet, or a high-fat diet containing genistein (HFG) for 6 months. During the study, indirect calorimetry, IP glucose tolerance tests, and behavioral analyses were performed. At the end of the study, plasma, liver, brain, and fecal samples were collected. The results showed that mice fed the HFG diet gained less weight, had lower serum triglycerides, and an improvement in glucose tolerance than those fed an HF diet. Mice fed the HFG diet also modified the gut microbiota that was associated with lower circulating levels of lipopolysaccharide (LPS) and reduced expression of pro-inflammatory cytokines in the liver compared to those fed HF diet. The reduction in LPS by the consumption of genistein was accompanied by an improvement of the cognitive function. Genistein is able to regulate the gut microbiota, reducing metabolic endotoxemia and decreasing the neuroinflammatory response despite the consumption of a HF diet.

Preventive effect of genistein on AOM/DSS-induced colonic neoplasm by modulating the PI3K/AKT/FOXO3 signaling pathway in mice fed a high-fat diet

Colon cancer is one of the most prevalent malignancies in the world. The incidence of colon cancer is on the rise, which seriously affects people's physical and mental health. Genistein, the predominant isoflavone found in soy products, has been shown to inhibit carcinogenesis in animal models. However, the molecular mechanisms involved in genistein preventing carcinogenesis are still uncharacterized. This study was aimed to evaluate the preventive effect of genistein-rich diet on azoxymethane/dextran sulfate sodium (AOM/DSS) induced colon cancer in high-fat mice. Contrasted with control group, the addition of genistein can ward off colon cancer. The expression of COX-2, TNF-α and FRAT1 mRNA has decreased in genistein-treated group. Furthermore, genistein-rich diet has inhibited the viability of colon cancer via regulating the expression of PI3K, FOXO3 and AKT. It has been suggested that genistein prevent the colonic neoplasms growth by modulating the PI3K/FOXO3/AKT signaling pathway in mice fed with a high-fat diet.

Effect of genistein addition to equine sperm freezing extender

Cryopreservation negatively affects equine sperm quality post-thaw: an excessive release of reactive oxygen species (ROS) and increased protein tyrosine phosphorylation (known as cryocapacitation) have been demonstrated in frozenthawed equine sperm diminishing its lifespan. The objective of this study was to determine the possible beneficial effect of genistein addition (a ROS scavenger and protein tyrosine kinase inhibitor) to a commercial freezing extender. Equine sperm were frozen in the presence of different genistein concentrations ranging from 0 to 800 μM. After thawing, the sperm viability (eosinnigrosin), acrosome integrity using peanut agglutinin (PNA), protein tyrosine phosphorylation (PY) and motility were assessed at times 0 and 1 hr. In addition PY was studied in fresh and frozen sperm incubated in Modified Whitten’s (MW) medium with 25 mM Bicarbonate (MW+Bic). Genistein did not affect the viability, total or progressive motility or acrosome status of frozenthawed equine sperm. Immediately after thawing, positive PY staining in the equatorial band was observed and genistein addition did not exert any change in PY. Fresh sperm incubated in MW+Bic showed a significant increase in PY staining along the tail compared to frozen-thawed sperm. In our study sperm viability immediately post-thaw was 58.25% ± 5.35 and progressive motility 14.46% ± 5.7 (mean ± S.E.M.) and genistein did not improve the post-thaw quality of equine sperm. In addition, cryopreservation did not induce PY immediately post-thaw or enhanced frozen-thawed equine sperm susceptibility to PY induction.

Anti-inflammatory, anti-osteoclastic, and antioxidant activities of genistein protect against alveolar bone loss and periodontal tissue degradation in a mouse model of periodontitis.

Genistein, a dietary polyphenol primarily found in soy products, has beneficial effects on bone. However, the effect of genistein on inflammatory periodontal destruction has not been investigated in detail. We explored whether genistein protects against lipopolysaccharide (LPS)/ligature-induced periodontitis in mice. We also examined the effect of genistein on LPS-stimulated inflammatory and oxidative stress using RAW 264.7 macrophages and human gingival fibroblasts (hGFs). The results from μCT and histological analyses revealed that intraperitoneal injection of genistein (20 mg/kg body weight) daily for three weeks inhibited LPS-mediated alveolar bone loss and periodontal tissue degradation. The administration of genistein also inhibited osteoclast formation and the expression of inflammation-related molecules in the inflamed region of mice with periodontitis. Treatment with 30-70 μM genistein significantly prevented osteoclast differentiation in receptor activator of nuclear factor κB ligand- or LPS-stimulated macrophages by suppressing the expression of osteoclast-specific molecules. The addition of genistein led to a dose-dependent inhibition of the expression of inflammation-related molecules both in LPS-stimulated macrophages and hGFs. In addition, genistein at 50 μM protected hGFs from LPS-mediated stresses such as mitochondrial impairment and cellular ROS accumulation. However, such protection was significantly diminished by combined treatment with 25 nM bafilomycin A1, a chemical autophagy inhibitor. Collectively, our results indicate that genistein protects against inflammatory periodontal damage by regulating autophagy induction and inhibiting osteoclast activation, the production of inflammation mediators, and mitochondrial oxidative damage. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2510-2521, 2017.

WPŁYW ZAWARTOŚCI Nmin NA NODULACJĘ ROŚLIN ŁUBINU ŻÓŁTEGO (Lupinus luteus L.) W OBECNOŚCI Bradyrhizobium lupini

A two-factor pot experiment was carried out in the years 2009-2011 under a permanent plastic-covered tunnel greenhouse at the Mochelek Experimental Station of the University of Technology and Life Sciences in Bydgoszcz (POLAND). The subject of research was the traditional cultivar of yellow lupin, Mister. The design of the experiment included 5 levels of Nmin content in the plant growth medium: from 4.45 mg Nmin.kg-1.d.w.– very low content to 22.5 mg Nmin.kg-1d.w. – very high content. Seeds of yellow lupin were inoculated with agar inoculum containing Bradyrhizobium lupini with or without the addition of genistein. Along with the plant growth, gradual decrease in the mean content of Nmin was observed, especially in the nitrogenous form until the stage of full bloom of lupin plants. No significant effect of the Nmin content was found on the initiation of the nodulation process, number and dry weight of nodules, independently of seed inoculation with Bradyrhizobium lupini and application of genistein. Average or higher content of Nmin in the medium was the reason of the lack of survival in symbiotic bacteria Bradyrhizobium lupini capable of nodulation in the medium after lupin harvest. Only in the case of low and very low Nmin content in the medium, the addition of genistein to bacterial inoculum did affect the increase in the survival of symbiotic bacteria after lupin harvest.

Involvement of a Novel Genistein-Inducible Multidrug Efflux Pump of &lt;i&gt;Bradyrhizobium japonicum&lt;/i&gt; Early in the Interaction with &lt;i&gt;Glycine max&lt;/i&gt; (L.) Merr

The early molecular dialogue between soybean and the bacterium Bradyrhizobium japonicum is crucial for triggering their symbiotic interaction. Here we found a single large genomic locus that is widely separated from the symbiosis island and was conspicuously induced within minutes after the addition of genistein. This locus (named BjG30) contains genes for the multidrug efflux pump, TetR family transcriptional regulator, and polyhydroxybutyrate (PHB) metabolism. The induction of BjG30 by genistein was competitively inhibited by daidzein, although both genistein and daidzein are soybean-derived inducers of nodulation (nod) genes. Such a differential expression pattern is also observed in some legume-derived flavonoids, which structurally differ in the hydroxy/deoxy group at the 5-position. In addition, not only did the induction start far in advance of nodW and nodD1 after the addition of genistein, but the levels showed distinct concentration dependence, indicating that the induction pattern of BjG30 is completely different from that of nod genes. The deletion of genes encoding either the multidrug efflux pump or PHB metabolism, especially the former, resulted in defective nodulation performance and nitrogen-fixing capability. Taken together, these results indicate that BjG30, and especially its multidrug efflux pump, may play a key role in the early stage of symbiosis by balancing the dual functions of genistein as both a nod gene inducer and toxicant.

Genistein promotion of osteogenic differentiation through BMP2/SMAD5/RUNX2 signaling.

To investigate the effects of Genistein on the osteogenic related gene expression profiles during osteoblastic differentiation of human bone marrow mesenchymal stem cell (hBMSC) cultures, the hBMSCs were cultured under osteogenic differentiation medium with the addition of Genistein (10-8∼10-5 M) for 12 days. The cell proliferation was measured by BrdU incorporation, while the osteoblastic differentiation in hBMSC cultures was assessed by cellular alkaline phosphatase (ALP) activity. The cell apoptosis was determined by caspase 3/7 activation. GEArray Q series human osteogenesis gene array was used to analyze large-scale gene expression in Genistein-treated hBMSC cultures compared to the control group. Quantitative real-time RT-PCR, small interfering RNA (siRNA), and western blot analysis were used to confirm the microarray data in five representative transcripts. Genistein (10-8∼10-6 M) dose- and time-dependently increased cell proliferation and cellular ALP activity, but had no significant effect on cell apoptosis in hBMSC cultures. The 96-gene array analysis indicated that 22 genes were upregulated more than 2-fold and 7 genes were downregulated at least 1.5-fold. The expressions of bone morphogenetic proteins (BMPs), small mothers against decapentaplegic homologs (SMADs), and Runt-related transcription factor 2 (RUNX2) were concomitantly increased under Genistein treatment while insulin-like growth factor 2 and inhibitory SMADs 6 and 7 expressions were significantly decreased. The results of the real-time RT-PCR had a correlation with the results of microarray analysis and were estrogen-receptor dependent. Specific gene siRNAs knock-down further confirmed the osteogenic effects of Genistein on BMP2, SMAD5 and RUNX2 protein expression. Genistein enhanced osteogenic differentiation in cultured hBMSCs mainly through the BMP-dependent SMADs and RUNX2 signaling.

The possibility of genistein as a new direct pulp capping agent

Genistein, kind of soy isoflavones, is well-known as natural ingredients and consumed as health foods and supplements. They are expected to improve renal function. They have high-affinity to estrogen receptor β expressed predominantly in bone tissue, they prevent osteoporosis specifically and safely. We examined whether genistein can be a new direct capping agent. In this study, we examined the effect of genistein for the proliferation and differentiation of rat dental pulp cells in vitro and the ability of tertiary dentin formation in vivo. As a result, rat dental pulp cells with genistein were increased activity of ALPase and showed alizarin red positive-staining. Calcification-related genes expression has been confirmed by the addition of genistein. From in vivo study, high quality of tertiary dentin formation and minor pulp reaction were observed. From these findings, it was suggested that genistein may be useful agent for direct pulp capping.