大豆异黄酮对去卵巢大鼠抗氧化及骨形态学影响的研究

目的 研究大豆异黄酮(SI)对去卵巢大鼠抗氧化及骨形态学的影响.方法 70只雌性SD大鼠按血清总胆固醇水平随机分为7组:高脂、雌激素,低、中、高剂量SI干预、假手术及正常对照组.对前5组摘除双侧卵巢后恢复1周开始干预,实验周期为12周,灌胃给药,每周称重1次,分别在去卵巢及处死时(12周)抽取尾静脉血,实验结束后取内脏及骨骼标本,分别测定血清碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶活性(GSH-Px)及骨密度等指标,共63只大鼠完成实验.结果 SI干预组股骨重量高于高脂与正常对照组,干预可提高AKP、GSH-Px酶活性.高剂量干预对维持大鼠股骨密度的作用与雌激素相似.SI干预可缓解因去卵巢造成的骨丢失.结论 大豆异黄酮对去卵巢大鼠的抗氧化、骨代谢酶活性及骨转化存在一定影响,适量干预能逆转因去势造成的骨密度下降.考虑到植物雌激素与哺乳动物的雌激素受体(ER)结合能力低下,采用SI在临床开展干预的剂量与远期效果尚待进一步研究.     

金雀异黄酮对去卵巢大鼠脑抗氧化作用的影响

目的研究金雀异黄酮对去卵巢大鼠大脑抗氧化作用的影响,为使用植物雌激素防治女性更年期后老年性痴呆提供依据。方法40只3月龄雌性SD大鼠,随机分为假手术组、去卵巢对照组、金雀异黄酮组和苯甲酸雌二醇组,给予相应手术和治疗。术后6周分析大鼠额叶、颞叶、海马和基底前脑的SOD和MDA值。结果假手术组颞叶和海马的SOD值低于基底前脑(P<0.05);额叶和颞叶的MDA值高于海马和基底前脑(P<0.05);去卵巢对照组基底前脑的SOD值显著低于假手术组、金雀异黄酮组和苯甲酸雌二醇组(P<0.05),MDA值显著高于假手术组、金雀异黄酮组和苯甲酸雌二醇组(P<0.05);假手术组、金雀异黄酮组及苯甲酸雌二醇组之间SOD值和MDA值差异无统计学意义(P>0.05)。结论去卵巢对照组大鼠基底前脑抗氧化能力显著降低,氧化损伤程度显著升高,金雀异黄酮替代治疗可增加去卵巢大鼠基底前脑的抗氧化能力,降低基底前脑的氧化损伤,可用于女性更年期后老年性痴呆的防治。     

金雀异黄酮对去卵巢后不同时间大鼠抗氧化作用的影响

目的研究雌激素剥夺后大鼠血清抗氧化作用的变化及金雀异黄酮(Gs)的治疗作用,探讨使用Gs进行雌激素替代治疗的可行性和最佳时间点。方法40只3月龄健康雌性SD大随机分为去卵巢对照组(OVX)、假手术组(sham)、金雀异黄酮组(Gs组)及苯甲酸雌二醇组(EB组),给予相应的手术和药物替代治疗。在术后第1、2、4、6周检测和分析血清超氧化物歧化酶(SOD)和丙二醛(MDA)的水平。结果术后第1～2周,各组大鼠血清SOD和MDA都逐渐上升,至第2周时达高峰,以后sham组、Gs组及EB组逐渐恢复到术后第1周水平,且各组之间均无显著性差异。但OVX组血清SOD活性持续下降,至术后第6周显著低于其它组(P<0.05);MDA值不断升高,在术后第4周(P<0.01)和第6周(P<0.05)都显著高于其它组。结论去卵巢2周后大鼠血清SOD活性逐步降低,氧化损伤程度不断增高;Gs具有与EB相似的抗氧化作用,可用于更年期后女性氧化损伤相关疾病的防治;雌激素替代疗法宜及早进行。     

大豆异黄酮对去势大鼠脂质过氧化及肝超微结构的影响

目的 研究大豆异黄酮(soybean isoflavone,SI)对去势大鼠脂质过氧化及肝超微结构的影响.方法 70只雌性SD大鼠按血清总胆固醇水平随机分为7组:高脂、雌激素,低、中、高剂量SI干预、假手术及正常对照组.前5组摘除双侧卵巢,恢复1周后开始干预,实验周期为12周,灌胃给药,每周称重1次,分别在去卵巢、给药4、8周、处死时抽取尾静脉血,实验结束后取肝脏标本,分别测定胆固醇(TC)、甘油三酯(TG)、血清高、低密度脂蛋白胆固醇(HDL-C、LDL-C)、脂蛋白(a)及抗氧化酶活性等.结果 SI干预组LDL-C水平显著低于高脂组但仍高于正常对照组;干预可使脂蛋白(a)水平发生变化但对HDL-C影响不显著;高剂量SI能缓解因去卵巢造成的肝细胞超微结构破坏,维护其完整性,全视野下以高剂量干预组与正常组结构最为接近,高脂组镜像损坏明显.结论 SI对去卵巢大鼠的脂蛋白(a)、脂质过氧化及肝超微结构存在影响,高剂量持续干预能降低去卵巢大鼠的LDL-C,并能维持肝超微结构.     

大豆异黄酮对大鼠血管平滑肌细胞增殖和表型转化的影响

目的:观察大豆异黄酮对大鼠血管平滑肌细胞增殖和表型转化的影响。方法:采用酶消化法原代培养大鼠血管平滑肌细胞,传至第3～6代,加入含不同浓度(0、0.1、0.2、0.4、0.8μmol/L)大豆异黄酮的DMEM培养基作用24 h,采用MTT法检测大鼠血管平滑肌细胞增殖率,采用RT-PCR法检测平滑肌22α(SM22α)mRNA、骨桥蛋白mRNA的表达,采用Western blot法检测SM22α、骨桥蛋白的表达。结果:0.1～0.8μmol/L的大豆异黄酮随浓度增加其抑制大鼠血管平滑肌细胞增殖的作用增强(P<0.05);SM22αmRNA及SM22α蛋白表达增加,骨桥蛋白mRNA及骨桥蛋白表达降低,各组间有明显差异(P<0.05)。结论:大豆异黄酮抑制大鼠血管平滑肌细胞增殖,同时使大鼠血管平滑肌细胞从合成表型向收缩表型转化。     

大豆异黄酮对去势雌大鼠缺血再灌注脑组织细胞凋亡及calpain表达的影响

目的: 研究大豆异黄酮对大鼠脑缺血再灌注后缺血脑组织神经细胞凋亡和calpain表达的影响,探讨大豆异黄酮的神经保护作用。方法: 成年雌性大鼠36只,随机分为药物组和对照组,药物组再随机分为小剂量和大剂量组。所有动物切除双侧卵巢,1周后各组每日给予大豆异黄酮和生理盐水,1个月后建立大脑缺血再灌注模型,原位末端标记检测神经细胞凋亡,采用原位杂交技术观察calpain mRNA表达情况,同时比较各组calpain活性变化。结果: 大豆异黄酮组凋亡细胞数明显减少(P<0.05),神经细胞凋亡指数降低,calpain mRNA表达明显减少,calpain活性下降。药物大剂量组与小剂量组比较无明显差异。结论: 大豆异黄酮使脑缺血再灌注后神经细胞凋亡减少,可能通过下调calpain mRNA表达,降低calpain的活性发挥神经保护作用。     

大豆异黄酮及主要活性成分Genistein对大鼠卵巢LRH-1基因表达的影响

目的 观察大豆异黄酮(soy isoflavones,SI)及主要活性成分Genistein对初老大鼠卵巢及体外培养卵巢颗粒细胞中肝受体类似物-1(liver receptor homolog-1,LRH-1)基因表达的影响,初步探讨大豆异黄酮作用于卵巢的分子机制.方法 采用自然老化法建立围绝经期大鼠动物模型,分别给予低剂量(50mg/kg)、中剂量(158 mg/kg)、高剂量(500 mg/kg)的SI灌胃处理8 w.分别采用原位杂交和RT-PCR检测卵巢组织中LRH-1 mRNA的表达.采用大鼠孕马血清促性腺激素(pregnant mare serum gonadotrophin,PMSG)皮下注射法建立促卵泡发育模型,分离、培养颗粒细胞24 h后,给予大豆异黄酮主要活性成分Genistein (0、0.1、1、5、10、100 μmol/L)以及雌激素受体(estrogen receptor,ER)阻断剂ICI182、780(1μmol/L)继续处理细胞24 h,采用RT- PCR方法检测卵巢颗粒细胞中LRH-1 mRNA的表达情况.结果 各剂量SI处理组卵巢LRH-1 mRNA的表达(低剂量0.563±0.037,中剂量0.926±0.127和高剂量1.223±0.134),与围绝经期模型组(0.460±0.082)相比均显著增加(P＜0.05).1～10 μmol/L的Genistein作用卵巢颗粒细胞24 h,LRH-1 mRNA的表达(分别为0.844±0.042,0.879±0.056,0.882±0.079)与空白对照组(0.678±0.052)相比均增加,结果具有统计学意义(P＜0.05).ER阻断剂ICI182,780预处理细胞30 min后再给予Genistein继续处理细胞24 h后,发现1～10 μmol/L的Genistein上调LRH-1表达作用,并未受ER阻断剂的影响.结论 一定剂量的大豆异黄酮可明显上调衰老卵巢LRH-1 mRNA的表达,1～10 μmol/L的Genistein可上调体外培养大鼠卵巢颗粒细胞中LRH-1 mRNA的表达,可能并非通过经典ER介导的.     

大豆异黄酮保护糖尿病肾病大鼠肾脏炎症及氧化应激损伤的研究

目的研究大豆异黄酮(SI)保护糖尿病肾病(DN)大鼠肾脏炎症及氧化应激损伤的研究。方法用链脲佐菌素腹腔内注射建立大鼠DN模型,并随机分为对照组、DN组和SI组,每组20只。模型构建8周后处死大鼠,肾脏称重计算肾体比,HE法检测各组肾脏病理改变;采用化学比色法检测血清和肾脏超氧化物歧化酶(SOD)和丙二醛(MDA)活性、总抗氧化能力(T-AOC)和晚期氧化蛋白产物(AOPPs)水平;ELISA法测定血清和肾脏TNF-α、IL-Iβ、IL-6水平。结果苏木素-伊红(HE)染色观察到,SI可以有效缓解肾小管损伤、肾小球系膜和基底膜的增生情况,减轻肾小管上皮水肿。与DN组相比,SI组SOD活性和T-AOC水平显著升高,MDA、血清AOPPs水平降低;血清TNF-α、IL-Iβ和IL-6含量显著降低(P<0.05)。结论大豆异黄酮具有抗炎抗氧化的作用,能够有效保护糖尿病肾病大鼠的肾脏中氧化应激及炎症反应。     

大豆异黄酮抑制大鼠前列腺增生及其作用机制

目的观察大豆异黄酮(SI)对良性前列腺增生(BPH)大鼠体内性激素、生长因子及细胞凋亡相关基因的影响,探讨SI对BPH的防治及作用机制。方法选择SPF级健康成年雄性SD大鼠100只,随机分为正常对照(NC)组、BPH模型组、低剂量[6 mg/(kg·d)]SI组、中剂量[12 mg/(kg·d)]SI组及高剂量[24 mg/(kg·d)]SI组,每组各20只。灌胃给药4周后麻醉处理大鼠,腹主动脉取血,制备血清;完整摘取大鼠前列腺组织。称重测定前列腺组织湿质量,计算前列腺指数(PI);酶联免疫吸附法(ELISA)测定大鼠血清雌二醇(E2)、睾酮(T)水平;免疫质印迹法(Western blotting)检测前列腺组织Fas、Fas L、Bax、Bcl-2、表皮生长因子(EGF)及其受体EGFR的表达。结果与NC组比,BPH组前列腺组织湿质量及PI均较明显增加(P<0.05);血清E2及T水平均显著升高(P<0.05);前列腺组织中Fas L、Bcl-2、EGF及EGFR表达水平均显著升高(P<0.05),而Fas和Bax表达水平均显著降低(P<0.05)。与BPH组比,中、高剂量SI组前列腺组织湿质量及PI均明显减少(P<0.05);血清E2及T水平均显著下降(P<0.05);前列腺组织中Fas L、Bcl-2、EGF及EGFR表达水平均显著降低(P<0.05),而Fas和Bax表达水平显著升高(P<0.05);其中,中剂量SI组各指标变化较低、高剂量组更显著(P<0.05)。结论 SI对BPH具有较好的抑制作用,其中中剂量SI作用效果更佳,其作用机制可能与调节机体性激素水平,下调生长因子及其受体表达以及调控细胞凋亡基因表达有关。     

大豆异黄酮的选择性雌激素受体调节剂作用

近年来 ,选择性雌激素受体调节剂和大豆异黄酮的研究不断深入 ,逐步证实大豆异黄酮在不同组织可以表现为雌激素激动剂和 (或 )拮抗剂的作用 ,本文将对大豆异黄酮的选择性雌激素受体调节剂的作用、作用机理以及应用和发展前景等进行概述。     

大豆异黄酮减轻反式脂肪酸诱导的大鼠动脉粥样硬化

目的观察大豆异黄酮减轻反式脂肪酸诱导的大鼠动脉粥样硬化的作用。方法将大鼠随机分为普通饲料组、反式脂肪酸组、大豆异黄酮组和反式脂肪酸+大豆异黄酮组,用试剂盒分别测量血中TG、HDL、LDL、IL-6和TNF-α的含量;培养牛主动脉内皮细胞,分为对照组、反式脂肪酸组和反式脂肪酸+大豆异黄酮组,Western blot法检测牛主动脉内皮细胞内NF-κB的磷酸化,同时检测细胞间黏附因子-1(ICAM-1)与血管细胞黏附因子(VCAM-1)蛋白表达。结果与普通饲料组比较,反式脂肪酸组血TG、LDL、IL-6和TNF-α明显升高(P0.05),HDL显著降低(P0.05)。大豆异黄酮干预后,血中TG、LDL、IL-6和TNF-α均降低(P0.01),HDL显著升高(P0.01)。大豆异黄酮能降低反式脂肪酸诱导的NF-κB磷酸化(P0.01);与对照组比较,大豆异黄酮能使反式脂肪酸诱导的ICAM-1和VCAM-1的表达降低(P0.01)。结论大豆异黄酮可减轻反式脂肪酸诱导的大鼠动脉粥样硬化作用。     

大豆异黄酮的药理功效研究进展

大豆异黄酮是大豆生长过程中形成的次生代谢产物,具有多种生物活性;近年来,大豆异黄酮已成为大豆最引人注目功能成分之一,也是药理学研究的热点之一。该文介绍大豆异黄酮在抗肿瘤、防治心血管疾病、抗炎、免疫调节、雌激素样作用等药理功效方面的研究进展及发展前景。     

Effects of a soybean milk product on feto-neonatal development in rats

Since estrogenic pollutants and phytoestrogens can cause the disorder of the reproductive system, the effects of a soybean milk product (Vegemil? containing 162 ppm isoflavones) on the feto-neonatal development, including male reproductive function, were investigated. Pregnant rats were fed the soybean milk (5% or 100% in drinking water) from gestational day (GD) 6 to parturition or to post-natal day (PND) 56. Specifically, the rats were divided into 4 groups: the control group (drinking water), the GD5% group (5% soybean milk during only the GD period), the GDPND5% group (5% soybean milk during the GD and PND periods), and the GD-PND100% group (100% soybean milk instead of water during the GD and PND periods). During the gestational, lactational, and developmental periods, the reproductive and developmental parameters of dams and offspring were observed. Feeding soybean milk did not affect the birth and physical development of both male and female offspring. At PND57, the weights of the testes and epididymides of F1 males significantly increased by feeding a high concentration of the soybean milk (GDPND100%). In addition, feeding of the soybean milk during both the GD and PND periods (GD-PND5% and GDPND100%) enhanced the sperm counts and motility. The results indicate that soybean milk is safe for embryos, fetuses, and offspring, and improves the post-generational development of male reproductive function.     

Molecular cloning and characterization of a new Japanese cedar pollen allergen homologous to plant isoflavone reductase family

BACKGROUND: Japanese cedar (Cryptomeria japonica) pollen is a major cause of seasonal pollinosis, and more than 10% of Japanese people suffer from this allergic disorder. However, only two major pollen allergens, Cry j 1 and Cry j 2, have been identified and exclusively characterized. OBJECTIVE: The aim of this study was to explore and identify important Japanese cedar pollen allergens other than Cry j 1 or Cry j 2. METHODS: C. japonica cDNA library was immunoscreened by rabbit antiserum raised against a partially purified cedar pollen allergen fraction. An isolated cDNA clone was inserted into a glutathione S-transferase (GST)-tagged Escherichia coli expression vector to obtain recombinant GST fusion protein. Non-fusion recombinant protein was purified by glutathione Sepharose affinity chromatography in conjunction with factor Xa cleavage of the GST moiety. IgE-binding ability of the recombinant protein was then evaluated by western blot analysis and enzyme-linked immunosorbent assay (ELISA). RESULTS: The cDNA encodes 306 amino acids with significant sequence similarity to those of plant isoflavone reductase-like proteins, which include a recently identified birch pollen allergen Bet v 5. Western blot analysis demonstrated that recombinant protein was recognized by cedar pollinosis patient IgE. In contrast to Bet v 5 being reported as a minor allergen, the recombinant protein exhibited 76% IgE binding frequency (19/25) against pollinosis patients. CONCLUSION: Here we identified the third member of Japanese cedar pollen allergen homologous to isoflavone reductase. Its high IgE-binding frequency implicates that the isoflavone reductase homologue might be an additional major pollen allergen in C. japonica.     

大白鼠皮下植入米非司酮埋植剂血药浓度和抑制异位子宫内膜效果的观察

目的观察大鼠皮下植入米非司酮埋植剂后的血药浓度变化及抑制大鼠子宫内膜异位症的效果。方法取体质量170～190g Wister雌性正常大鼠，分别在正常大鼠和动物模型大鼠（实验组5组和对照组）皮下埋植单根0．75cm、1．5cm、3．0cm和2根、3根3cm长米非司酮埋植剂（外径2．5mm），对照组植入2根3．0cm不含药的埋植剂。正常大鼠给药1个月．定期取血用HPLC法测血药浓度。动物模型大鼠给药3个月．用MRI测定异位子宫内膜体积并计算抑制率。结果不同剂量的米非司酮皮下埋植剂皆在植入体内7d时，血药浓度达到最高：随着给药剂量加大，血药浓度峰值也趋向增高，但7d后血药浓度逐渐下降，14d后除3根剂量组的血药浓度显著较高外（约0．18～0．20μg／ml），其他较低剂量组之间的血药浓度差异无统计学意义（约0．10～0．13μg/ml）。大鼠皮下植入埋植剂的剂量与血药浓度峰值呈线性关系。动物模型皮下植入1根1．5cm、3．0cm或2根3．0cm长埋植剂1个月时，异位内膜抑制率分别为18．6％±17．3％、31．5％±12．7％、72．2％±12．3％，与对照组比较差异有统计学意义，并且不再随剂量加大而显著提高抑制效果。结论 米非司酮皮下埋植剂作为治疗子宫内膜异位症的长效制剂是可行的．     

The effect of superoxide dismutase on the blood lipoprotein level of rabbits in the dynamics of traumatic shock

Changes of the lipoprotein (LP) level in blood in the abdominal aorta and the portal and hepatic veins were studied in experiments on rabbits with Cannon's traumatic shock. The LP level was determined by spectrophotometry. The blood LP level changed depending on the stage of the shock. Administration of superoxide dismutase (an enzyme whose biosubstrates are only products of a free-radical process) in the initial period of shock has a normalizing effect on the level of the main LP classes.     

Development of an LC-MS/MS-based analytical method for quantification of soybean allergen Gly m 4 in soybean grains and processed foods

Gly m 4 is one of important soybean allergens, which has property to cross-react with birch pollen allergen. However, a universal method for quantifying soybean allergen is unavailable. We aimed to develop an LC-MS/MS-based method for Gly m 4 quantification. A recombinant Gly m 4 (rGly m 4) was used to plot a standard curve and conduct a recovery test. The recovery of rGly m 4 from soybean grain extracts was 128% and the intra- and inter-day coefficients of variation were 12.8% and 13.2%, respectively. The method had a limit of quantification of 0.1?µg/mL. Using a validated method, Gly m 4 content was found to range from 309 to 421?µg/g in soybean grains and 2.7–67?µg/g in soybean processed foods. The newly developed method demonstrated sufficient reproducibility, sensitivity and accuracy, and might be helpful for individuals with soybean allergy to control Gly m 4 intake from food products.     

Multi-agent systems simulating the physiological role of plasmic membrane

This paper presents our multi-agent framework for modelling and simulating the physiological role of plasmic membrane specially the 'endocytosis by receptors' and the 'exocytosis' mechanisms. As the dysfunction of a plasmic membrane may cause several illnesses (the family hypercholesterolemia, the dilation and the permeability of blood vessel, the allergic reactions as the asthma, rums of hays, hives, etc.), the objective of our multi-agent framework is to be a virtual world of cellular and molecular biology while simulating the physiological role of plasmic membrane in order to help its specialists to better understand, to good interpret and to warn changes of cell interaction and cell adaptation with its environment via its plasmic membrane. The architecture of our multi-agent framework is composed of a set of reactive agents. These agents use an ontology (named OntoCell) to communicate and to represent their knowledge. OntoCell describes the knowledge of cellular biology. Our framework is implemented with a multi-agent platform (named DIMA).     

大豆异黄酮酊干预Ⅱ度烫伤模型小鼠创面的愈合

背景：雌激素具有促进创面愈合的作用,大豆异黄酮能与雌激素受体结合,具有明显的雌激素样效应。 目的：探索大豆异黄酮酊的制备、观察其对小鼠烫伤创面愈合的影响。 方法：用冷浸渍法制备大豆异黄酮酊,筛选最佳有效浓度。用昆明小鼠造Ⅱ度烫伤模型,随机分为6组。药物组分别应用0.100,0.361,1.000,3.610 g/L的大豆异黄酮酊局部涂抹;溶剂对照组应用体积分数75%乙醇局部涂抹;空白对照组不作任何处理。从第 3 天起,隔天记录创面面积、计算未愈合创面占原始创面的百分比,于造模后第 3,9,14 天取创面组织做病理切片,观察形态学改变和创面愈合情况。 结果与结论：大豆异黄酮酊的最佳效应浓度是3.61 g/mL。在创伤后第 5,7,9,11,13 天,大豆异黄酮酊组的未愈合创面占原始创面的百分比明显小于溶剂对照组和空白组(P < 0.05)。病理组织切片显示,第 9 天,大豆异黄酮酊组的成纤维细胞明显多于溶剂对照组和空白组,第 14 天,大豆异黄酮酊组的表皮生长情况比溶剂对照组和空白组好。表明以75%乙醇为溶剂制备大豆异黄酮酊是可行的;  3.61 g/L的大豆异黄酮酊对小鼠创面的愈合具有明显的促进用。