Effects of PVP K30 on Aqueous Solubility and Dissolution Propertiesof Daidzein

AimTo study effects of PVP K30 on the aque ou s solubility and dissolution properties of daidzein. Methods To measure the aqueous solubility and dissolution rates of daidzein in three diffe rent states (within solid dispersions, physical mixtures and as a pure drug) and investigate drug-polymer interactions in the solid state using X-ray powder d iffraction and fourier-transform infrared spectroscopy. Results The negative values of the Gibbs free energy and enthalpy of transfer explaine d the spontaneous transfer of daidzein from phosphate buffer solution (PBS) to a solution of PVP in PBS. X-ray powder diffraction patterns showed that the drug was in the amorphous state (ratio of the drug ∶ PVP<1∶5) when dispers ed i n PVP K30. The infrared spectra indicated there exist interactions between the O H of daidzein and the C=O of PVP K30. Conclusionthe dispersion of daidzein in PVP K30 considerably enhances the solubility and dissolution rat e.

植物雌激素大豆苷原(daidzein)对结肠癌LoVo细胞生长的影响

目的 了解植物雌激素大豆苷原 (daidzein)对结肠癌细胞生长的影响。方法 取对数生长期的人结肠癌细胞株LoVo细胞 ,以不同浓度的大豆苷原处理细胞 2 ,3,4和 6d ,然后应用MTT法测定细胞的生长率。结果 大豆苷原在高浓度 10 0 μmol/L时在体外对LoVo有明显的抑制作用 (P <0 .0 1) ,而在低浓度 <5 μmol/L时对LoVo细胞又具有促增殖作用。

Response surface optimization of microwave-assisted extraction for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii

Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol（1∶15,v/v）for 30 min,and then microwave irradiation for 11 min at a power of 600 W.Coupling the extraction process with HPLC-fluorescence presented good recovery,satisfactory precision,and good linear relation.Compared with a method from the Chinese Pharmacopoeia,the proposed method enables higher extraction efficiency and more accurate analytical results.It can be of potential value in quality assessment of Radix Puerariae thomsonii medicinal materials.

Neuroprotective effects of daidzein on focal cerebral ischemia injury in rats

Daidzein, a plant extract, has antioxidant activity. It is hypothesized, in this study, that daidzein exhibits neuroprotective effects on cerebral ischemia. Rat models of middle cerebral artery oc- clusion were intraperitoneally administered daidzein. Biochemical and immunohistochemical tests showed that superoxide dismutase and nuclear respiratory factor 1 expression levels in the brain tissue decreased after ischemia and they increased obviously after daidzein administra- tion; malondialdehyde level and apoptosis-related cysteine peptidase caspase-3 and caspase-9 immunoreactivity in the brain tissue increased after ischemia and they decreased obviously after daidzein administration. Hematoxylin-eosin staining and luxol fast blue staining results showed that intraperitoneal administration of daidzein markedly alleviated neuronal damage in the ischemic brain tissue. These findings suggest that daidzein exhibits neuroprotective effects on ischemic brain tissue by decreasing oxygen free radical production, which validates the afore- mentioned hypothesis.

Dietary Daidzein Enhances Antiapoptotic Effect of 17β-Estradiol (E_2) on Breast Cancer MCF-7 Cells

Objective： To investigate whether dietary daidzein interact with endogenous 17β-Estradiol （E2） to give rise to additive or inhibitory effects on proliferation and apoptosis in breast cancer cells. Methods： Cell cycle distribution and apoptosis induction were analyzed by using flow cytometry when breast cancer cell lines MCF-7 were cotreated with daidzein （1, 5 μmol/L） and E2 （0.1-10 nmol/L） for 5 days. Whether daidzein could alter E2-modulated mRNA expression of estrogen receptor alpha （ERα）, estrogen receptor beta （ERI3） and ERβ-estrogen response element （ERE） dependent transcription was investigated by RT-PCR and luciferase induction assays. The effects of daidzein on E2-modulated expression of proapoptotic p53, bax and antiapoptotic bcl-2 at both mRNA and protein levels were also investigated by RT-PCR and Western blot. Results： Daidzein enhanced the antiapoptotic effect in an Ea dose-dependent manner, but had no effect on E2-induced proliferation. Daidzein antagonized E2-induced ERβ mRNA expression and ERβ-ERE dependent transcription. In addition, daidzein only antagonized E2-upregulated expression of p53 and bax, but had no effect on E2-upregulated expression of bcl-2. Conclusion： Daidzein enhances the antiapoptotic effect of E2 on breast cancer cells by inhibiting E2-mediated p53-bax proapoptotic pathway. These results suggest that dietary daidzein may enhance deleterious effect of endogenous E2 in hormone-dependent breast cancer.

Dietary supplementation with a high dose of daidzein enhances the antioxidant capacity in swine muscle but experts pro-oxidant function in liver and fat tissues

Background： Although isoflavones are natural dietary antioxidants, they may have toxicological effects. This study aimed to evaluate the redox system in tissues of finishing pigs by supplementation with high dose of daidzein（640 mg/kg）.Results： The supplementation of high dose of daidzein for 64 d increased the activity of superoxide dismutase and total antioxidant capacity in longissimus muscle but down-regulated the expression of reactive oxygen species（ROS）-producing enzyme NADPH oxidase-2 and cyclooxygenase-2. In contrast, high-level supplementation with daidzein exerted pro-oxidant changes in back fat, abdominal fat, liver, and plasma, as reflected by increased contents of malondialdehyde, a lipid peroxidation product, in these tissues. Furthermore, daidzein supplementation resulted in higher expression of ROS-producing enzymes, including NADPH oxidase-1 and cyclooxygenase-1in liver, 5-lipoxygenase（5-LOX） in backfat and NADPH oxidase-2 both in abdominal fat and backfat. The supplementation of daidzein did not affect meat quality parameters in longissimus muscle, including marbling score,eye muscle areas, intramuscular fat, shear force, drip loss, p H and meat color.Conclusions： This experiment suggests that dietary supplementation of finishing pigs with daidzein at a high dose level improves redox status in muscle but exerts pro-oxidant effect in liver and fat tissues.

RP-HPLC method for the simultaneous determination of daidzein,genistein and formonetin in Solanum Lyratum Thunb

A rapid method for the simultaneous determination of daidzein,genistein and formonetin in solanum Lyratum Thunb by high performance liquid chromatography(HPLC)was developed.Separation was achieved on a Diamonsil C18 column(250 mm×4.6 mm,5 μm)with isocratic elution,using a mobile phase of methanol-tetrahydrofuran-water(44∶3∶53,v/v).The wavelength was set at 260 nm and column was maintained at 35 ℃.The linear ranges of daidzein,genistein and formonetin were 1.0-40.0,0.1-4.0 and 0.1-4.0 μg/mL,respectively.The average recoveries were between 98.4% and 101.3%.This method could be used for the quality control of Solanum lyratum Thunb due to its simplification,reliability,rapidity and excellent precision.

Effect of daidzein on milk yield,components of milk and hormones in serum of dairy cows

Four Holstein dairy cows in the 7th month of late lactation were sampled, and self-control method was used. Daidzein was added to the concentrated feed at the dose of 10mg/kg during the treatment stage. Results showed that daidzein lessened the declining tendency of milk yield in late lactation and increased the milk yield to some extents. Lactoprotein was evidently increased (P<0.01), while milkfat was not affected evidently. The level of T4 in serum was evidently increased (P<0.05); the levels of GH, PRL, E2 and TSH in serum were prone to rising while T3 was prone to decreasing . It was concluded that daidzein could indirectly affect milk yield and components of milk by regulating the levels of intrinsic hormones.

Attenuating effect of daidzein on polychlorinated biphenyls-induced oxidative toxicity in mouse testicular cells

The attenuating effect of daidzein (DAI) on oxidative toxicity induced by Aroclor 1254 (A1254) was investigated in mouse testicular cells. Cells were exposed to A1254 alone or with DAI. The oxidative damage was estimated by measuring malondialdehyde (MDA) formation, superoxide dismutase (SOD) activity and glutathione (GSH) content. Results show that A1254 induced a decrease of germ cell number, an elevation in thiobarbituric acid reactive substances (TBARS) but a decrease in SOD activity and GSH content. However, simultaneous supplementation with DAI decreased TBARS level and increased SOD activity and GSH content. Consequently, dietary DAI may restore the intracellular antioxidant system to attenuate the oxidative toxicity of A1254 in testicular cells.

3′-Daidzein sulfonate sodium protects against memory impairment and hippocampal damage caused by chronic cerebral hypoperfusion

3′-Daidzein sulfonate sodium（DSS） is a new synthetic water-soluble compound derived from daidzein,a soya isoflavone that plays regulatory roles in neurobiology.In this study,we hypothesized that the regulatory role of DSS in neurobiology exhibits therapeutic effects on hippocampal damage and memory impairment.To validate this hypothesis,we established rat models of chronic cerebral hypoperfusion（CCH） by the permanent occlusion of the common carotid arteries using the two-vessel occlusion method.Three weeks after modeling,rat models were intragastrically administered 0.1,0.2,and 0.4 mg/kg DSS,once a day,for 5 successive weeks.The Morris water maze test was performed to investigate CCH-induced learning and memory deficits.TUNEL assay was used to analyze apoptosis in the hippocampal CA1,CA3 regions and dentate gyrus.Hematoxylin-eosin staining was performed to observe the morphology of neurons in the hippocampal CA1,CA3 regions and dentate gyrus.Western blot analysis was performed to investigate the phosphorylation of PKA,ERK1/2 and CREB in the hippocampal PKA/ERK1/2/CREB signaling pathway.Results showed that DSS treatment greatly improved the learning and memory deficits of rats with CCH,reduced apoptosis of neurons in the hippocampal CA1,CA3 regions and dentate gyrus,and increased the phosphorylation of PKA,ERK1/2,and CREB in the hippocampus.These findings suggest that DSS protects against CCH-induced memory impairment and hippocampal damage possibly through activating the PKA/ERK1/2/CREB signaling pathway.

Genistein and daidzein reduced chlorpyrifos induced damage in PC12 cell

The present study preliminarily evaluated neurotoxicity injuries induced by chlorpyrifos in PC12 cell,which were used as a model for nervous cell system.In cultured PC12 cell,application of soy isoflavones(genistein,daidzein,monomer and mixture)significantly reduced chlorpyrifos induced toxicity,a widely used pesticide,and resulted in a better cell survival rate.Treatments with isoflavones reduced malondialdehyde content,reactive oxygengeneration and acetylcholine level in medium,and maintained mitochondrial membrane potential integrity.Daidzein enhanced endogenous antioxidant system in PC12 cell with an increasing in superoxide dismutase perunit activity.Genistein reduced acetylcholine content in the medium.Daidzein and genistein showed different effects,and their combined effect were greater than individual.In conclusion,soy isoflavones as an antioxidant and neuroprotectant,enhanced choline metabolism,which effectively mitigated disadvantageous influence of PC12 cell caused by chlorpyrifos.

The Effect of Intra-cerebral Microinjection with Daidzein on the Plasma LH Level in the Castrated Boars

Daidzein (DA) was microinjected into mediobasal hypothalamus (MBH) and nucleus ventrome-dialis thalami (VM) of the castrated male Goettingen Mini pigs (n = 9, 10 μl·animal-1, 8 pgDA·μl-1 and plasma luteinizing hormone (LH) levels were compared between the prior and the posterior treatment. The result showed that LH levels after the cerebral administration (ad) tended to increase compared to the levels before ad. In MBH, LH levels in 4 cases (4/5), rose and were not changed in 1 case at 0.5 - 2 hours after ad compared to those before ad. There were no significant changes at 2.5 hours after ad. When it was injected in VM, LH levels in 3 cases (3/4) rose, and were not changed in 1 case after ad compared to those before ad. In the control, there were no changes in plasma LH levels between the pre-and post-treatment except 1 case in MBH. This study suggested that DA could up-regulate LH secretion through hypothalamus level in the castrated boars.

Neuroprotective effect and mechanism of daidzein in oxygen-glucose deprivation/reperfusion injury based on experimental approaches and network pharmacology

Background:Daidzein,phytoestrogens derived from the Pueraria lobata(Willd.)Ohwi root used in traditional Chinese medicine,has a wide range of biological activities,including antioxidant,anti-inflammatory,and neuroprotection.However,the neuroprotective role of daidzein in oxygen-glucose deprivation/reperfusion injury and its underlying mechanism are still unknown.Methods:In this study,we used pheochromocytoma cells induced by oxygen-glucose deprivation and reperfusion to study the potential effect in the protection of the nerve cells.Then,we used molecular docking simulation and network pharmacology to predict the possible targets and pharmacological pathways of daidzein.Western blot was used to verify the expression of target proteins with or without adding the inhibitors.Results:After daidzein treatment,cell vitality had an upward trend(P<0.05)and the release of lactate dehydrogenase had a downward trend(P<0.01)in dose-dependent compared with the model group by exposure to oxygen-glucose deprivation and reperfusion.Several core targets were analyzed through network pharmacology and molecular docking including catalase,peroxisome proliferator-activated receptor gamma,vascular endothelial growth factor A,interleukin-6,tumor necrosis factor,nitric oxide synthase 3,prostaglandin-endoperoxide synthase 2,and RAC-alpha serine/threonine kinase 1.These results suggest that catalase may be a first-ranked target for the neuroprotective role of daidzein.Gene Ontology enrichment analysis indicated the pathways mainly contained molecule metabolic process,while Kyoto Encyclopedia of Genes and Genomes enrichment analysis focus on pathways in terms of inflammation such as tumor necrosis factor signal pathway.Then,Western blot results showed that daidzein had a significant increase on the expression of protein catalase(P<0.01).Daidzein reversed catalase level alterations after oxygen-glucose deprivation reperfusion injury in a dose-dependent manner which was consistent with the catalase antagonists-based experiments.Conclusion:These outcomes provide new insights into the neuroprotective effect and mechanism of daidzein in oxygen-glucose deprivation/reperfusion injury.

Protective effects of 3′-daidzein sulfonate sodium on chronic hepatic injury by regulating the T lymphocyte subsets in mice

OBJECTIVE To explore the protective effects of 3′-daidzein sulfonate sodium on chronic hepatic injury induced by carbon tetrachloride(CCl4)in mice and investigate the mechanism about regulating the T lymphocyte subsets.METHODS Healthy Kunming male mice were randomly divided in 5groups:control group,model group,bifendate positive control group(2.5mg·kg-1),low and high dose 3′-daidzein sulfonate sodium groups(0.1and 0.3mg·kg-1).The chronic hepatic injury mice were made by intraperitoneal injection of 10% CCl4 plant oil solution twice a week,and sustained for six weeks.At the same time,the mice were treated with normal saline,bifendate(2.5mg·kg-1)and 3′-daidzein sulfonate sodium(0.1and 0.3mg·kg-1),respectively by ig administration once a day and continued for six weeks.After the last administration,the mice blood and liver were taken.Using automatic biochemical analyzer survey the activity of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in serum.Flow cytometry was used to detect T lymphocyte subsets,enzymes analysis technique was used to observe the liver function(CD3+,CD4+and CD8+),HemateinEosin stain was used to explore the changes in liver morphology.RESULTS The level of ALT and AST in the serum in model group mice increased significantly.While the level of ALT and AST in the serum in 3′-daidzein sulfonate sodium(0.1 or 0.3mg·kg-1,ig)groups mice decreased compared with the model group(P<0.05).And compared with the model group,the ratios of CD3+,CD4-CD8-and CD4+/CD8+ decreased,and the ratios of CD8+ increased in 3′-daidzein sulfonate sodium groups(0.1 and 0.3mg·kg-1,ig;P<0.05).CONCLUSION 3′-daidzein sulfonate sodium have a protective effect on CCl4-induced liver injury mice,and it can result in the changes of T lymphocyte subsets,which may be one of the factors leading to hepatic injury by CCl4.

3′-Daidzein sulfonate sodium improves mitochondrial functions after cerebral ischemia/reperfusion injury

3′-Daidzein sulfonate sodium is a new synthetic water-soluble compound derived from daidzein（an active ingredient of the kudzu vine root）. It has been shown to have a protective effect on cerebral ischemia/reperfusion injury in rats. We plan to study the mechanism of its protective effect. 3′-Daidzein sulfonate sodium was injected in rats after cerebral ischemia/reperfusion injury. Results showed that 3′-daidzein sulfonate sodium significantly reduced mitochondrial swelling, significantly elevated the mitochondrial membrane potential, increased mitochondrial superoxide dismutase and glutathione peroxidase activities, and decreased mitochondrial malondialdehyde levels. 3′-Daidzein sulfonate sodium improved the structural integrity of the blood-brain barrier and reduced blood-brain barrier permeability. These findings confirmed that 3′-daidzein sulfonate sodium has a protective effect on mitochondrial functions after cerebral ischemia/reperfusion injury, improves brain energy metabolism, and provides protection against blood-brain barrier damage.

大豆黄酮影响青春期小鼠乳腺发育的潜在机制

[目的]以大豆黄酮(DZ)作用于青春期雌性小鼠,探讨其通过上调雌二醇(E2)水平和雌激素受体β(ERβ)受体表达影响青春期小鼠乳腺发育的机制,为DZ及其他植物雌激素类在畜牧生产和临床应用提供依据。[方法]选取48只2周龄C57BL/6雌性小鼠,随机均分为3组:对照组,腹腔注射无菌的PBS;DZ处理组,腹腔注射DZ(100 mg·kg^(-1));阳性对照组,腹腔注射E2(0.2 mg·kg^(-1)),每组16只。分别在饲养至21和28 d,每组随机选取8只,采集血液,测定血液中E2含量,处死后摘取乳腺,测定乳腺指数并检测乳腺组织中E2含量。取同侧部位乳腺制作组织学切片,免疫组化检测核增殖抗原蛋白(PCNA)的表达,Western blot法分析乳腺组织内细胞周期蛋白D1(cyclin D1)和D3(cyclin D3)、ERβ蛋白表达水平。[结果]与对照组相比,DZ处理显著增加饲养21~28 d小鼠体重及采食量;增加21 d小鼠血液和乳腺组织中E2含量,但未达到显著水平(P>0.05);显著增加28 d小鼠血液和乳腺组织中E2含量(P<0.05);E2处理血液中E2含量均极显著增加。DZ处理对饲养21 d小鼠乳腺指数影响较小,显著增加饲养28 d小鼠乳腺指数(P<0.05);E2处理具有相同趋势(P<0.05)。HE染色结果显示,DZ和E2处理均极显著增加饲养21和28 d小鼠乳腺组织小叶内的导管数(P<0.01)和乳腺组织PCNA阳性细胞数(P<0.01)。Western blot结果显示,DZ及E2处理均可上调小鼠乳腺组织cyclinD1、cyclinB1、ERβ蛋白的表达,显著上调饲养28 d小鼠cyclinD1、cyclinB1、ERβ蛋白的表达(P<0.05)。[结论]DZ处理可促进青春期小鼠乳腺发育,促进乳腺细胞增殖和分裂,这种作用可能与内源性E2水平升高及ERβ受体有关。

植物雌激素大豆黄酮对断奶期小鼠乳腺退化的减缓作用研究

旨在研究植物雌激素大豆黄酮(DZ)对断奶期小鼠乳腺退化的影响。选取48只孕鼠,随机分为对照组(CON组),腹腔注射无菌PBS;大豆黄酮组(DZ组),每天每kg体重腹腔注射DZ 100 mg,连续处理1周。孕鼠正常分娩后,在哺乳期第10天强制断奶。各组分别在断奶第1、3、7天选取8只小鼠取血后处死,取所有乳腺组织,称重计算乳腺指数;ELISA法测定血液中雌二醇(E2)、催乳素(PRL)、胰岛素样生长因子(IGF-Ⅰ)的浓度;常规法制作乳腺组织切片,进行HE染色;Western blot及荧光定量PCR(RT-qPCR)分析乳腺组织内促凋亡因子Bax和抗凋亡因子Bcl-2的蛋白与基因表达水平。结果:与CON组相比,DZ组增加了断奶1 d母鼠的乳腺指数,但降低断奶3 d、7 d的乳腺指数;与CON组相比,断奶1 d时,DZ组极显著提高母鼠血清中PRL和IGF-Ⅰ浓度(P<0.01),断奶3 d时DZ组极显著提高了PRL和E2浓度(P<0.01),显著提高断奶7 d母鼠血清中E2浓度(P<0.05);HE染色结果显示,与CON组相比,DZ组显著降低断奶母鼠乳腺组织中脂肪细胞面积比(P<0.05),提高腺泡的面积比(P>0.05);与CON组相比,DZ组极显著上调断奶期乳腺组织Bcl-2蛋白的表达水平(P<0.01),极显著下调断奶1 d Bax基因的表达水平(P<0.01),断奶3 d时极显著下调Bax蛋白与基因的表达水平(P<0.01),断奶7 d时极显著上调乳腺中Bax和Bcl-2蛋白的表达水平(P<0.01),在断奶1 d和3 d,Bcl-2/Bax比值均极显著升高(P<0.01)。本研究表明,DZ处理可升高断奶母鼠血清中IGF-Ⅰ、PRL及E2浓度,上调乳腺中Bcl-2/Bax比值,抑制细胞凋亡,降低脂肪细胞所占面积比,提示DZ可能通过抑制母鼠乳腺细胞凋亡,延缓乳腺早期退化。

大豆连作及其根茬腐解物对大豆根系分泌物中异黄酮类物质—黄豆甙元(daidzein)的影响

本试验通过收集正茬、重茬7年、重茬7年并经MB-生物制剂调控和加入根茬腐解物(20g kg-1土)的正茬、重茬、调控的大豆根系分泌物,采用高效液相色谱检测方法,对根系分泌物中的大豆异黄酮—黄豆甙元(daidze in)进行了检测。结果在正茬根系分泌物中没有检测到该物质,在重茬、调控根系分泌物中检测到了该物质,其检测出的量为重茬>调控。在加入根茬腐解物的各处理的根系分泌物中均检测到了大豆异黄酮,其检测出的量为重茬>正茬>调控。加入根茬腐解物的处理与不加根茬腐解物的相应的处理相比,在其根系分泌物中检测到的大豆异黄酮的量均有显著提高。

Synthesis of Novel Phosphorylated Daidzein Derivatives and ESI Investigation on Their Non-Covalent Complexes with Lysozyme

Daidzein （7,4＇-dihydroxyisoflavone） was phosphorylated by a modified Atherton-Todd reaction. The structures of the five target product, were determined by X-ray, IR, NMR and ESI-MS. Electrospray ionization results show that in the gas phase all the phosphorylated daidzein derivatives could form non-covalent complexes with the protein lysozyme, while non-covalent complexes were not detected in the mixed solution of daidzein with lysozyme. Relative affinity of every non-covalent complex was obtained according to its different decomposition orifice voltage.

Supplementing daidzein in diets improves the reproductive performance, endocrine hormones and antioxidant capacity of multiparous sows

Certain hormones play important roles in modulating mammalian reproductive behaviour.Daidzein is a well-known isoflavonic phytoestrogen that possesses oestrogenic activity.This study was conducted to probe the effects of daidzein supplementation in gestation diets on the reproductive performance in sows.A total of 120 multiparous sows(Landrace x Yorkshire)were randomly assigned to 2 groups(n=60)and fed either a base diet(control)or one containing 200 mg/kg daidzein during gestation.We discovered that daidzein supplementation significantly increased the total number of piglets born per litter and number of piglets born alive per litter(P<0.05),decreased the farrowing time(P<0.05)and increased the serum oestrogen and progesterone concentrations(P<0.05)at 35 d of gestation.Moreover,serum immunoglobulin G(IgG)concentration and superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)activities were higher in the daidzein-treated group than in the control group at35 d of gestation(P<0.05).Daidzein increased the serum SOD activity and total anti-oxidative capacity(T-AOC)at 85 d of gestation(P<0.05).Interestingly,daidzein elevated the expression levels of the sodium-coupled neutral amino acid transporter 1(SLC38 A1)and insulin-like growth factor 1(IGF-1)genes in the placenta(P<0.05).These results suggest that daidzein ingestion could improve sow reproductive performance by changing serum hormones,elevating anti-oxidative capacity and upregulating critical functional genes in the placenta.