Estrogen Receptor α and β Expressions in Hypothalamus-pituitary-ovary Axis in Rats Exposed Lactationally to Soy Isoflavones and Bisphenol A

Objectives This paper aims to investigate the uterotrophic activities of lactational exposure to combination of soy isoflavones （SIF） and bisphenol A （BPA） and to examine estrogen receptor α （ERα） and estrogen receptor β （ERβ） expressions in hypothalamus-pituitary-ovary axis and uterus.Methods Maternal rats that were breeding about 8 litters were randomly divided into four groups with seven dams in each group.Dams in different treatment groups received corn oil （control）,150 mg/kg BW of SIF,150 mg/kg BW of BPA or combination of 150 mg/kg BW of SIF and 150 mg/kg BW of BPA,respectively,from postnatal day 5 to 11 （PND5-11） by gavage.On PND12 and PND70,10 female litters were killed and hypothalamus,pituitary,ovary and uterus were collected.ERα and ERβ expressions in these organs were detected with Western blotting assay.And vaginal opening time and estrus cycle were examined in animals fed for PND70.Results On PND12,the relative uterine weight of rats treated with ISF or BPA or their combination was significantly higher than that of untreated rats （P〈0.05）.But the relative uterine weight of rats in the co-exposure group was slightly lower than that in the group only exposed to SIF or BPA.On PND 70,however,the relative uterine weight in each treatment group was not statistically different from that in the control group （P〈0.05）.Vaginal opening time and estrus cycle in groups treated with SIF or BPA or their combination were similar to those in the control group （P〈0.05）.Exposure to SIF or BPA or their combination could up-regulate or down-regulate ERα and ERβ expressions in hypothalamus,pituitary,ovary and uterus on PND12 and PND70.These regulation patterns for ERα and ERβ were different in different organs at different time points.Conclusion Lactational exposure to ISF or BPA or their combination could induce uterotrophic responses in neonate rats,which disappeared in later life.But these data fail to suggest a possibility for synergic actions between SIF and BPA.It was also demonstrated that the uterotrophic effects of SIF and BPA exposure might,at least,involve modification of ERα or ERβ expressions in the hypothalamus-pituitary-ovary axis.

Effects of Dietary Soy Isoflavones on Feed Intake,Growth Performance and Digestibility in Juvenile Japanese Flounder(Paralichthys olivaceus)

An 8-week feeding trial was conducted to investigate the effects of dietary soy isoflavones on feeding intake,growth performance,and digestion of juvenile Japanese flounder(Paralichthys olivaceus).Four isonitrogenous(49% crude protein) and isoenergetic(20.1 MJ kg-1) diets were formulated to contain four graded levels of soy isoflavones,namely,0,1,4 and 8 g soy isoflavones in 1 kg of diet.Each diet was randomly fed to triplicate tanks of fish(Initial average weight:2.58 g ± 0.01 g),and each tank was stocked with 35 fish.No significant difference was observed among diets with levels of 0,1 and 4 g kg-1 soy isoflavones in feed intake,weight gain,feed efficiency ratio(FER),proximate composition of fish whole body and apparent digestibility coefficients(ADC) of nutrients and energy(P>0.05).However,high dietary soy isoflavones level(8 g kg-1) significantly depressed weight gain,FER,whole-body crude lipid content of fish and ADC of nutrients(P<0.05).These results indicate that high level of dietary soy isoflavones(above 4 g kg-1) significantly depresses growth responses and FER of Japanese flounder.However,as the content of soy isoflavones in soybean meal is around 1 to 3 g kg-1,the adverse effects might be neglected when soybean products are used as a fish feed ingredient.

Consumption of soybean,soy foods,soy isoflavones and breast cancer incidence:Differences between Chinese women and women in Western countries and possible mechanisms

Breast cancer is one of the most lethal diseases world-wide.However,there is a large difference in breast cancer incidence among Caucasian,Hispanic,African and Asian(e.g.Chinese)women with Caucasian women being the highest and Asian women being the lowest.It has been suggested that the dietary factors may account for approximately 50%of the breast cancer.One of such dietary components which are typical to Asian but not Caucasian diet is soy foods.A number of epidemiological studies have suggested that increasing soy consumption could be related to the decreased risk of occurrence and/or mortality of breast cancer.In this review,we first described briefly different types of soy products and their nutritional functions and consumption.Then,we described briefly soybean isoflavones,i.e.genistein(GEN),daidzein,glycitein,and presented several lines of evidence to demonstrate the possible association of soy flavone food consumption with incidence and prognosis of breast cancer;finally,we summarized several possible molecular mechanisms,including the effects of GEN as an agonist of ER,epigenetic and genome-wide effects,activation of peroxisome proliferator-activated receptors,induction of apoptosis and stimulation of autophagy,involved in the chemo-preventive effects of GEN on breast cancer.©2013 Beijing Academy of Food Sciences.Production and hosting by Elsevier B.V.All rights reserved.

Effects of soybean isoflavones on reproductive parameters in Chinese mini-pig boars

Background： Soybean isoflavones are structurally similar to mammalian estrogens and therefore may act as estrogen agonists or antagonists. However, it has not been determined if they have any negative effects on reproductive parameters in male livestock. Therefore, the objective of this study was to evaluate the effects of soybean isofiavones on male reproduction using Chinese mini-pig boars as a model. Fifty Xiang boars were randomly divided into five groups and fed diets containing 0, 125, 250, or 500 ppm soybean isoflavones or 0.5 ppm diethylstilbestrol for 60 days. Results： Dietary supplementation with 250 ppm of soy isoflavones markedly increased the testis index （P 〈 0.05）, fructose content （P 〈 0.05）, and o-glycosidase content in testicular tissue （P 〈 0.01）, as well as increased the number of viable germ cells （P 〈 0.01） and the level of Bcl-2 protein （P 〈 0.01）. However, 500 ppm of soybean isoflavones significantly reduced both testis and epididymis indexes （P 〈 0.0S） and lactate dehydrogenase levels （P 〈 0.01）, as well as reduced serum LH and testosterone levels （P 〈 0.05）. High levels of soybean isoflavones also increased malondialdehyde levels （P 〈 0.05）, as well as increased the numbers of early and late apoptotic germ cells （P 〈 0.01） and the level of Bax proteins （P 〈 0.05） in the testis. Conclusions： The results of this study indicate that consumption of soy isoflavones at dietary levels up to 250 ppm did not adversely affect reproductive parameters in Chinese mini-pig boars whereas higher levels of soy isoflavones may adversely affect male reproduction.

Subchronic Toxicity Study on Soy Isoflavones in Rats

Objective To investigate the subchronic toxicity of soy isoflavones （SIF） in male rats. Method Fifty Sprague-Dawley rats were randomly divided into 5 groups, 10 rats per group. SIF were given to rats in different groups by gavage at dose of 0, 0.2, 0.5, 1.5, and 4.5 g/kg bw, respectively for 13 weeks. Clinical manifestations, body weight, and food consumption were observed weekly. At the end of the study, urinalysis, hematology, clinical chemistry, total testosterone, and follicle-stimulating hormone were tested, and histopathological examinations were performed. Results No mortality, ophthalmic abnormalities or treatment-related clinical signs were identified during the study. As compared with the control group, significantly lower body weights and food consumption were observed in 1.5 and 4.5 g/kg bw groups. In clinical chemistry tests, triglyceride was significantly decreased and high-density lipoprotein cholesterol was significantly increased in all SIF-treated groups. Total testosterone levels were significantly lower in 0.50, 1.50, and 4.5 g/kg bw dose groups than in the control group. Microscopic examination showed that the mammary glands exhibited hyperplasia and excreted latex in rats of the 4.5 g/kg bw group. No changes attributable to treatment of SIF in other parameters were found. Conclusion SIF at high dosages caused significant endocrine disruption in male rats. The no observed adverse effect level （NOAEL） of SIF to male rats in this study is considered to be 0.20 g/kg bw.

Effects of Soy Isoflavones on the Sex Hormone Level and Pathological Changes in the Perimenopausal Syndrome Animal Model

[Objectives] To observe the effect of soy isoflavones on the perimenopausal syndrome model of female rats,and to find its mechanism,in order to provide experimental evidence for the clinical treatment of menopausal syndrome. [Methods]50 female SD rats were randomly divided into five groups,with 10 rats in each group,respectively,for the normal control group,sham operation group,model group,diethylstilbestrol group,soy isoflavone treated group,except the normal control group and sham operation group,the other groups were ovariectomized rats,the formation model of perimenopausal rats treated with normal saline,diethylstilbestrol,soy isoflavone by gavage,normal control group and sham operation group were given the same dose of normal saline and soy isoflavone by gavage. After 6 weeks of continuous treatment,abdominal aorta was taken blood,the serum was isolated,and radioimmunoassay method to measure E_2( Eestradiol),P( Progesterone),T( Testosterone). And removal of the pituitary gland,uterine and adrenal prepared pathological section,experimental effect was observed by light microscopy,using immunohistochemical method to observe the brain pituitary FSH( Follicle-Stimulating Hormone) and LH( Luteinizing Hormone) levels. [Results] Radiation immune analysis results showed that soy isoflavone could significantly improve serum E_2 level in ovariectomized rats,compared with the model group,the statistical analysis showed significant difference( P < 0. 05),but the serum P and T level had little effect. The pathological results showed that( i) uterus: the uterine wall and endometrial thickness of soy isoflavone group was between that of model group and sham operation group,the surface epithelium was columnar,glands increased;( ii) pituitary gland: soy isoflavone ovariectomized group cells were between model group and normal group;( iii) adrenal gland: soy isoflavone group adrenal tissue did not change significantly. Immunohistochemical results showed that soy isoflavone group pituitary tissues of FSH and LH cells compared with the model group decreased significantly. [Conclusions] The soy isoflavones could make the uterus wall and endometria of perimenopausal syndrome animal model thicken,improve the serum level of E_2 in ovariectomized rats,and decrease the level of FSH and LH,but had little effect on the adrenal tissues,level of T and P.

Urine Metabonomic Analysis of Interventions Effect of Soy Isoflavones on Rats Exposed to Di-(2-ethylhexyl) Phthalate

Objective Di-(2-ethylhexyl) phthalate(DEHP) is a ubiquitous environmental contaminant.As an endocrine disruptor,it seriously threatens human health and ecological environmental safety.This study examines the impact of intervention with soybean isoflavones(SIF) on DEHP-induced toxicity using a metabonomics approach.Methods Rats were randomly divided into control(H),SIF-treated(A,86 mg/kg body weight),DEHP-treated(B,68 mg/kg),and SIF plus DEHP-treated(D) groups.Rats were given SIF and DEHP daily through diet and gavage,respectively.After 30 d of treatment,rat urine was tested using UPLC/MS with multivariate analysis.Metabolic changes were also evaluated using biochemical assays.Results Metabolomics analyses revealed that p-cresol glucuronide,methyl hippuric acid,N1-methyl-2-pyridone-5-carboxamide,lysophosphatidycholine [18:2(9 Z,12 Z)] {lyso PC [18:2(9 Z,12 Z)]},lyso PC(16:0),xanthosine,undecanedioic acid,and N6-acetyl-l-lysine were present at significantly different levels in control and treatment groups.Conclusion SIF supplementation partially protects rats from DEHP-induced metabolic abnormalities by regulating fatty acid metabolism,antioxidant defense system,amino acid metabolism,and is also involved in the protection of mitochondria.

Molecular docking studies of human MCT8 protein with soy isoflavones in Allan-Herndon-Dudley syndrome(AHDS)

Monocarboxylate transporter-8 （MCT8） is a specific thyroid hormone transporter, essential for the uptake of thyroid hormone into target tissues. Mutations in the MCT8 gene have been identified as the cause of Allan-Herndon-Dudley syndrome （AHDS）. It has been reported that soy isoflavones influence thyroid hormone system and can interact with thyroid hormone transporter proteins. Therefore, the present study aimed to find out whether soy isoflavones （genistein, daidzein and glycitein） can be used as a natural inhibitor to target MCT8 in AHDS. Docking studies were performed for soy isoflavones in order to evaluate their binding affinity to MCT8 protein using AutoDock4 （version 4.2.6） and AutoDock Vina. After docking, the ligands were ranked according to their binding energy and the best lead compound was selected based on the least binding energy. The docking results indicated that daidzein possesses the lowest binding energy against MCT8. Moreover, it was found that the residues PRO-338, HIS-341, and GLU-348 were involved in hydrogen bond interactions with genistein and daidzein. This study suggests that daidzein is a promising natural inhibitor to target MCT8 in AHDS.

Effect of daily supplementation of soy isoflavones on hot flashes and night sweats in African menopausal women

The aim of this study was to evaluate the effectiveness of a pharmaceutical grade isoflavone supplement (Inoclim?) 40 mg per capsule of soy extract rich in daidzin and genistin) on the number of hot flashes and night sweats in menopausal African women. This open, prospective, multicenter study involved 140 menopausal women with climacteric symptoms (i.e., a minimum of five hot flashes and/or moderate to severe night sweats per day). These patients received a daily 40-mg supplement of soy isoflavone contained in an Inoclim? capsule (Laboratoire Innotech International, France). The dose was increased to two capsules per day when there were more than five hot flashes or when the women could not sleep because of night sweats. The patients were followed up over a 4-month period and data were analyzed using EPI-InfoTM version 3.5.1. The outcome measure was fewer hot flashes and night sweats. The mean daily number of episodes of hot flashes was 5.7 at baseline and 1.8 at week 16, corresponding to a decrease of 68.4% (n = 93 patients at baseline and n = 66 at 16 weeks). The mean number of daily episodes of night sweats was 3.7 at baseline and 0.8 at week 16, corresponding to a decrease of 78.4% (n = 127 patients in the first week and n = 73 at 16 weeks). Over 80% of patients took a single one 40 mg capsule per day. The percentage of patients who adhered to the prescription was more than 95% during the 4-month follow-up period. These results confirm the effectiveness of a pharmaceutical grade isoflavone supplement (40 mg, rich in genistin and daidzin) on vasomotor symptoms in menopausal women.

异黄酮与β-伴大豆球蛋白的相互作用及其对蛋白结构和潜在致敏性的影响

探究异黄酮和β-伴大豆球蛋白(β-conglycinin,BCG)的相互作用机理及其对复合物结构和潜在致敏性的影响。利用荧光光谱、圆二色光谱分析2种大豆异黄酮(染料木素(genistein,Gen)、大豆苷元(daidzein,Dai))与BCG相互作用的猝灭类型、结合位点数、作用力类型和蛋白二级结构含量的变化,表征不同条件下制备的异黄酮-BCG复合物的结构,并利用酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)与消化产物免疫印迹检测其潜在致敏性。结果表明:活性异黄酮(Gen/Dai)对BCG的猝灭为静态猝灭,且相互作用力以疏水相互作用为主,结合位点数均接近1;并且与异黄酮相互作用诱导了BCG氨基酸微环境的极性增加,使蛋白肽链伸展,结构更为松散;ELISA与消化产物免疫印迹结果显示,与异黄酮结合后蛋白的潜在致敏性增强。本研究有助于科学认识异黄酮在食品复杂基质体系中对过敏蛋白致敏性影响机制,对其抗过敏的深度开发利用具有重要意义。

SIF干预肥胖大鼠下丘脑瘦素介导JAK/STAT信号通路的研究

通过观察SIF对肥胖大鼠下丘脑中瘦素介导的信号通路蛋白表达的影响,探讨SIF对肥胖大鼠的干预机制。采用高脂饲料饲喂大鼠,建立肥胖动物模型,并将肥胖大鼠分为4组,分别灌胃SIF,剂量为:对照组(Ⅰ组)0 mg·kg^(–1)、低剂量组(Ⅱ组)50 mg·kg^(–1)、中剂量组(Ⅲ组)150 mg·kg^(–1)、高剂量组(Ⅳ组)450 mg·kg^(–1),并设置正常大鼠基础组(Ⅴ组),饲喂基础饲料,并灌胃给予SIF处理等体积的羧甲基纤维素钠,连续4周。采用HE染色法观察下丘脑组织学结构变化,免疫组织化学SABC法检测下丘脑中瘦素介导的JAK/STAT信号转导通路蛋白OB-Rb(瘦素受体长型),JAK2(内源性酪氨酸蛋白激酶2),p-STAT3(磷酸化的信号转导与转录激活子3),SOCS3(细胞因子信号3抑制因子),NPY(神经肽Y)的表达水平。结果显示:高剂量组和基础组OB-Rb,JAK2,p-STAT3的阳性表达水平显著高于其他组(P<0.05),且前述蛋白均有随SIF的剂量增加而表达增多趋势。高剂量组和基础组的SOCS3和NPY表达水平显著较其他组低(P<0.05),但NPY在背内侧核和室旁核的表达随SIF的剂量增加而增多。OB-Rb,JAK2,p-STAT3,SOCS3和NPY均在下丘脑广泛表达,并随SIF的剂量增加而变化,提示SIF对瘦素介导的信号通路有一定作用并与机体能量的调节密切相关;SIF通过干预大鼠下丘脑瘦素JAK/STAT信号通路具有减重作用,并呈剂量依赖性。

饲粮中添加大豆异黄酮对安格斯肉牛生产及繁殖性能的影响

【目的】研究饲粮中添加不同水平大豆异黄酮对安格斯肉牛生长性能、血清生化指标、抗氧化能力、免疫性能及繁殖激素的影响,探讨大豆异黄酮在肉牛生产实践中的适宜添加量。【方法】选取40头体重相近、2~3胎次且健康的安格斯母牛,随机分为4组,每组10头。对照组牛饲喂基础饲粮,试验Ⅰ、Ⅱ、Ⅲ组牛分别在基础饲粮中添加10、20和40 mg/kg大豆异黄酮。试验始末对牛进行称重,记录每日采食量,计算平均日采食量(ADFI)、平均日增重(ADG)及料重比(F/G);第40天时,采集血液检测血清生化、抗氧化、免疫指标及繁殖激素水平。【结果】与对照组相比,(1)添加大豆异黄酮对安格斯肉牛ADFI、ADG及F/G均无显著影响(P>0.05)。(2)试验Ⅰ、Ⅱ组牛血清中总蛋白含量显著升高(P<0.05),谷丙转氨酶活性显著降低(P<0.05);试验Ⅲ组牛血清中总蛋白和球蛋白含量极显著升高(P<0.01),谷草转氨酶和谷丙转氨酶活性极显著降低(P<0.01);试验Ⅲ组牛血清中免疫球蛋白(IgA、IgM、IgG)及γ-干扰素水平极显著或显著升高(P<0.01;P<0.05)。(3)试验Ⅱ组牛血清中总抗氧化能力(T-AOC)及谷胱甘肽过氧化酶(GSH-Px)活性均极显著升高(P<0.01);试验Ⅲ组牛血清中T-AOC、GSH-Px及超氧化物歧化酶水平均极显著升高(P<0.01),丙二醛含量极显著降低(P<0.01)。(4)试验Ⅱ组牛血清中雌二醇水平显著升高(P<0.05);试验Ⅲ组牛血清中孕酮、雌二醇、促卵泡激素及促黄体生成素水平均极显著升高(P<0.01)。【结论】饲粮中添加大豆异黄酮可有效提高安格斯肉牛免疫功能和繁殖机能,其中40 mg/kg大豆异黄酮添加量效果最佳。

基于发酵过程的淡豆豉HPLC-ELSD特征图谱建立及指标性成分含量测定

目的:研究发酵过程中淡豆豉样品化学成分变化的特点,建立可用于淡豆豉质量评价的高效液相色谱-蒸发光散射检测法(HPLC-ELSD)特征图谱,并对淡豆豉中大豆异黄酮类和大豆皂苷类生物活性成分进行含量评价。方法:采用Agilent ZORBAX SB C_(18)色谱柱(150 mm×4.6 mm,3.5μm),流动相A为0.1%乙酸水溶液,流动相B为0.1%乙酸乙腈溶液,梯度洗脱,流速为1.0 mL·min~(–1),柱温为30℃,蒸发光散射检测器气体流量为1.6 L·min~(–1)、漂移管温度为60℃、喷雾器温度为30℃。基于选定的过程样品,确定经发酵后含量升高的共有峰,进而建立淡豆豉特征图谱。以淡豆豉特征图谱为对照图谱,对15批市售淡豆豉样品的HPLC-ELSD色谱图进行相似度评价,并对其10个共有峰峰面积进行正交偏最小二乘法-判别分析评价。采用HPLC-二极管阵列检测器法(HPLC-DAD)对淡豆豉中6个大豆异黄酮类物质进行含量测定;采用HPLC-ELSD对淡豆豉中3个B族大豆皂苷进行含量测定。结果:基于10批30个发酵过程样品,发现淡豆豉中存在多个含量升高的色谱峰;其中10个共有峰峰面积升高较显著,进而建立了淡豆豉的特征图谱;通过与对照品比对,指认了其中6个共有峰。根据市售淡豆豉样品的HPLC-ELSD特征图谱相似度是否大于0.95,可将15批市售淡豆豉分为优质淡豆豉和基本合格淡豆豉。10个共有峰峰面积的正交偏最小二乘法-判别分析结果与相似度分析结果一致。15批市售品中大豆苷元和染料木素总质量分数为0.12%±0.06%,大豆皂苷Ba、大豆皂苷Bb和大豆皂苷Bc的总质量分数为0.59%±0.09%;优质淡豆豉中大豆苷元和染料木素总质量分数为0.15%±0.07%,大豆皂苷Ba、大豆皂苷Bb、大豆皂苷Bc总质量分数为0.63%±0.10%,大豆皂苷Bb质量分数为0.46%±0.08%;基本合格淡豆豉中大豆苷元和染料木素总质量分数为0.09%±0.04%,大豆皂苷Ba、大豆皂苷Bb、大豆皂苷Bc总质量分数为0.56%±0.07%,大豆皂苷Bb质量分数为0.41%±0.06%。结论:建立的HPLC-ELSD特征图谱精密度、重复性、稳定性良好,能够体现淡豆豉中大豆皂苷与异黄酮多种组分的含量特点,可用于提升淡豆豉的质量控制水平。指标性成分含量测定方面,建议在《中华人民共和国药典》2020年版标准基础上,进一步提高大豆苷元和染料木素总量的限量标准,且可增加大豆皂苷Bb含量测定项。

大豆异黄酮对西农萨能奶山羊繁殖性能及其羔羊的影响

[目的]大豆异黄酮(SIF)是一种生物活性因子,影响激素的分泌和代谢。本试验旨在探讨在繁殖前短期补充SIF对西农萨能母山羊繁殖性能及其羔羊的影响。[方法]试验于配种前选取30只健康的泌乳奶山羊,随机分为3组:对照组(常规日粮)、SIF-1组(常规日粮+100 mg SIF)和SIF-2组(常规日粮+200 mg SIF),饲喂28天。[结果]SIF-1组和SIF-2组的奶山羊干物质采食量均显著高于对照组(P<0.05)。3组母羊血清雌二醇浓度在试验期间均随时间延长而显著升高(P<0.05)。SIF-1组和SIF-2组的奶山羊发情率与受胎率均高于对照组。SIF-1组的奶山羊繁殖率高于对照组和SIF-2组。SIF组的羔羊体长显著高于对照组,其中SIF-2组的羔羊体长最大(P<0.05)。[结论]奶山羊繁殖期短期补充SIF可以提高其繁殖性能,在饲喂常规日粮(其中精料补充料大豆异黄酮含量为148.6mg/kg)基础上,每只奶山羊每天补充100mgSIF更为理想。

饲料中大豆异黄酮检测方法的建立

[目的]建立用于检测饲料中大豆提取物的高效液相色谱方法(HPLC),为大豆异黄酮在畜牧业的合理使用提供支持。[方法]通过查阅文献,采用HPLC色谱技术,样品经甲醇溶液超声提取后,以乙腈-0.1%醋酸水为流动相梯度洗脱,260 nm波长检测。[结果]建立的HPLC检测方法,线性范围为0.2~20μg/mL,定量限可达0.197μg/mL,以2、15μg/mL的质量浓度进行添加回收,添加回收率为97.45%~102.54%。[结论]按建立的HPLC方法检测5%纯度的大豆异黄酮,可检测6种组分的含量分别为:大豆苷1.13%、大豆黄苷0.22%、染料木苷2.79%、大豆苷元0.55%、大豆黄素0.06%和染料木素0.29%,总黄酮含量为5.04%。

大豆异黄酮通过抑制Wnt/Ca2+信号通路减轻大鼠脑缺血再灌注引起的钙超载

目的 探讨大豆异黄酮(SI)减轻脑缺血再灌注(I/R)引起钙超载的作用机制。方法 将48只SD大鼠采用随机数法分为4组:假手术组(Sham组)、脑缺血再灌注模型组(I/R组)、病毒空载组(NC组)、Frizzled-2敲低组(Knock down组),12只/组。采用线栓法堵塞大脑中动脉2 h,再灌注24 h构建I/R模型。采用Western blot验证病毒敲低效率并检测Wnt/Ca2+信号通路相关蛋白Wnt5a、Frizzled-2和P-CaMKⅡ的变化。采用钙含量显色法检测各组缺血半暗带(IP)区钙离子浓度变化,HE检测各组IP区组织结构变化。另将72只SD大鼠采用随机数法分为3组:Sham组、I/R组、大豆异黄酮预处理组(SI组),24只/组。采用多普勒血流仪检测局部脑血流变化,TTC染色检测脑梗死体积,HE和尼氏染色检测IP区组织变化、免疫荧光检测ROS、Ca2+和细胞凋亡水平、流式检测细胞钙离子浓度,试剂盒检测血清MDA和SOD水平,Western blotting和免疫组化检测IP区Wnt5a、Frizzled-2和P-CaMKⅡ蛋白表达。结果 与I/R组比较,Knock down组钙离子浓度(P<0.001)、Wnt5a(P<0.05)、Frizzled-2(P<0.05)和P-CaMKⅡ(P<0.001)表达水平均降低;与I/R组比较,SI组钙离子浓度(P<0.05)、ROS和MDA水平(P<0.001)、细胞凋亡程度(P<0.001)、脑梗死体积(P<0.001)、Wnt5a、Frizzled-2和P-CaMKⅡ表达水平(P<0.05)均降低,SOD水平升高(P<0.001)。结论大豆异黄酮可能通过抑制Wnt/Ca2+信号通路减轻大鼠脑缺血再灌注引起的钙超载。

大豆异黄酮抑制Slit2/MAPK信号通路对牙周炎大鼠牙槽骨吸收和炎症反应的影响

目的:基于Slit同源物2(Slit2)/P38丝裂原活化蛋白激酶(MAPK)信号通路探讨大豆异黄酮(SIF)对牙周炎大鼠牙槽骨吸收和炎症反应的影响。方法:将大鼠分为对照组(Control)、模型组(Model)、SIF低剂量组(L-SIF,25 mg/kg)、SIF高剂量组(H-SIF,75 mg/kg),每组10只,除对照组外,其余各组使用丝线结扎大鼠上颌第一磨牙的牙颈部建立牙周炎模型。L-SIF组和H-SIF组大鼠灌胃相应剂量SIF,Control组和Model组灌胃等量生理盐水,1次/d,连续4周。给药结束后,ELISA检测大鼠血清Slit2、TNF-α、IL-6、IL-1β水平;Micro-CT扫描检测釉牙骨质界至牙槽骨嵴顶(CEJ-ABC)距离和骨矿物质密度(BMD)、骨体积分数(BV/TV),评估牙槽骨丢失情况;HE染色和抗酒石酸酸性磷酸酶(TRAP)染色评估组织炎症反应、骨吸收和破骨细胞活性;免疫组织化学法(IHC)检测牙周组织骨保护素(OPG)和核因子κB受体活化因子配体(RANKL)表达;Western blot检测牙周组织Slit2、P38 MAPK、p-P38 MAPK蛋白表达。结果:与Control组相比,Model组大鼠血清Slit2和TNF-α、IL-1β、IL-6水平、CEJ-ABC距离、牙周组织病理学损伤评分、破骨细胞数量、RANKL阳性表达、Slit2蛋白水平和p-P38 MAPK/P38 MAPK显著升高,BMD和BV/TV、牙周组织OPG阳性表达显著降低(均P<0.05);与Model组相比,L-SIF组和H-SIF组大鼠血清Slit2和TNF-α、IL-1β、IL-6水平、CEJ-ABC距离、牙周组织病理学损伤评分、破骨细胞数量、RANKL阳性表达、Slit2蛋白水平和p-P38 MAPK/P38 MAPK显著降低,BMD和BV/TV、牙周组织OPG阳性表达显著升高(P<0.05),且H-SIF组上述指标变化显著优于L-SIF组(P<0.05)。结论:SIF可抑制牙周炎大鼠牙槽骨吸收和炎症反应,改善牙周炎,其作用机制可能与抑制Slit2/P38 MAPK信号通路有关。

豆制品中大豆异黄酮的转化与富集研究进展

豆制品是日常生活中极为常见的食品,它们富含大豆异黄酮。这种物质进入人体后会转化为游离型大豆异黄酮苷元,具有预防骨质疏松、缓解女性更年期综合征、抗氧化与抗癌等积极效果,因此在食品保健领域具有极为广阔的应用前景。本文总结了豆制品中大豆异黄酮的转化与富集方法的研究进展,旨在为大豆异黄酮在食品保健领域中的创新与应用提供坚实的理论参考和技术支持。

超高效液相色谱法检测大豆异黄酮含量

目的:建立一种测定大豆中大豆异黄酮含量的高效检测方法,研究大豆中大豆异黄酮成分的差异。方法:采用超高效液相色谱法检测大豆中大豆异黄酮含量,在大豆样品中加入70%甲醇溶液,超声波法提取大豆异黄酮,色谱柱为BEHC C_(18)柱(150 mm×2.1 mm,1.7μm),流动相为乙腈-0.2%乙酸,检测波长为260 nm,柱温为30℃,流速为0.15 mL·min^(-1)。结果:4种大豆异黄酮成分的线性关系、精密度、稳定性良好,平均回收率为97.6%~98.6%。大豆中大豆异黄酮成分大豆苷含量最高,大豆苷元含量最低,具体含量为大豆苷352.0μg·g^(-1),染料木苷88.0μg·g^(-1),染料木素25.8μg·g^(-1),大豆苷元19.5μg·g^(-1)。结论:超高效液相色谱法操作简单、检测速度快、灵敏度高,可用于大豆中大豆异黄酮含量检测。

Systematic review of soy isoflavone supplements on osteoporosis in women

Objective:To clarify the effect of soy isoflavones on prevention of osteoporosis,and the effective dosage of soy isoflavones and its duration.Methods:Random control trials that investigated the association of soy isoflavones and osteoporosis were included in the meta-analysis by researching MEDLINE,EMBASE and the Chinese Biomedical Database up to October 2011.The RevMan software was used for all of the statistical analysis.Results:The present meta-analysis found that soy isoflavones significantly increased the bone mineral density by 54%and decreased the bone resorption marker urinary deoxypyridinoline(DPD) by 23%compared to baseline in women. Using random effects model,the effect of isoflavones on bone mineral density(BMD) regarding menopausal status and isoflavone dose revealed higher weighted mean difference changes were found in postmenopausal women and isoflavone dose above 75 mg/d.Subgroup analysis of trials with menopausal status,supplement type,isoflavone dose and intervention duration that used soy isoflavone extracts resulted in significant diflerent overall effect of DPD using by random effects model.Sensitivity analysis indicated that the effect of soy isoflavones on BMD and DPD was robust.Conclusions:The present meta-analysis reveals that soy isoflavone supplements significandy increase bone mineral density and decrease the bone,resorption marker urinary DPD.It shows no significant effect on bone formation markers serum bone alkaline phosphatase. The significant effect of soy isoflavones on BMD and urinary DPD is relative to menopausal status, supplement type,isoflavone dose and intervention duration.