Poly(chitosan-ester-ether-urethane) Hydrogels as Highly Controlled Genistein Release Systems

Polymeric hydrogels play an increasingly important role in medicine, pharmacy and cosmetology. They appear to be one of the most promising groups of biomaterials due to their favorable physicochemical properties and biocompatibility. The objective of the presented study was to synthesize new poly(chitosan-ester-ether-urethane) hydrogels and to study the kinetic release of genistein (GEN) from these biomaterials. In view of the above, six non-toxic hydrogels were synthesized via the Ring-Opening Polymerization (ROP) and polyaddition processes. The poly(ester-ether) components of the hydrogels have been produced in the presence of the enzyme as a biocatalyst. In some cases, the in vitro release rate of GEN from the obtained hydrogels was characterized by near-zero-order kinetics, without “burst release” and with non-Fickian transport. It is important to note that developed hydrogels have been shown to possess the desired safety profile due to lack of cytotoxicity to skin cells (keratinocytes and fibroblasts). Taking into account the non-toxicity of hydrogels and the relatively highly controlled release profile of GEN, these results may provide fresh insight into polymeric hydrogels as an effective dermatological and/or cosmetological tool.     

负载染料木素纳米脂质体的制备 及其体外抗炎活性研究

为了提高染料木素（Gen）的生物利用率以及为脂溶性生物活性物质在新型功能食品中的开发提供参考,采用薄膜分散法,以大豆卵磷脂和胆固醇为原料,制备了负载Gen的纳米脂质体（Gen@Lip）,并对其基本指标、结构、储存稳定性、体外释放特性和抗炎活性进行分析。结果表明：制备的Gen@Lip的Gen负载量为36%,Gen@Lip的平均粒径为（242.73±2.03）nm,多分散指数为0.32,Zeta-电位为（-16.33±1.16）mV;结构表征结果证实Gen被有效包埋在Gen@Lip中;Gen@Lip具有良好的储存稳定性;Gen@Lip能够有效地通过胃进入到肠道实现缓释作用;相较于游离的Gen,Gen@Lip对Raw 264.7细胞有更强的抑炎效果,且在使用范围（＜100 μmol/L）内无毒副作用。综上,所制备的Gen@Lip可提高Gen的生物利用率,并具有明显的抗炎效果。     

染料木黄酮抑制3T3-L1细胞成脂分化机制

目的：研究染料木黄酮是否通过雌激素受体介导影响3T3-L1前脂肪细胞成脂分化,并探讨其可能的机制。方法：以不同浓度染料木黄酮处理3T3-L1细胞,四甲基偶氮唑蓝（methyl thiazolyl tetrazolium,MTT）法测定细胞存活率;在3T3-L1前脂肪细胞成脂分化过程中分别加入不同浓度染料木黄酮、染料木黄酮和ICI182780（一种雌激素受体抑制剂）混合物及不同浓度雌二醇,油红染色观察分化结果,测定细胞甘油三酯（triglyceride,TG）含量;染料木黄酮、染料木黄酮和ICI182780混合物及不同浓度雌二醇处理诱导成熟后的脂肪细胞,测定培养液甘油含量;分别用染料木黄酮（30 μmol/L）、染料木黄酮（30 μmol/L）和ICI182780混合物干预细胞成脂分化,Western blotting法检测细胞外信号调节激酶（extracellular signal-regulated kinase,ERK）、p-ERK、腺苷酸活化蛋白激酶（adenosine monophosphate activated protein kinase,AMPK）、p-AMPK、激素敏感性脂肪酶（hormone sensitive lipase,HSL）、脂肪酸合成酶（fatty acid synthetase,FAS）蛋白表达量。结果：3T3-L1细胞存活率随染料木黄酮浓度的升高而降低,50～200 μmol/L染料木黄酮能抑制细胞生长（P＜0.01）;染料木黄酮能负向调节成脂分化后细胞胞浆内TG含量,在0～50 μmol/L浓度范围内呈剂量依赖关系,高剂量雌二醇（100 nmol/L）能下调TG含量;染料木黄酮能促进成熟脂肪细胞脂解,提高细胞培养液甘油浓度;染料木黄酮（30 μmol/L）能上调p-AMPK、HSL蛋白表达,下调FAS蛋白表达（P＜0.01）,对ERK、p-ERK、AMPK表达量无明显影响（P＞0.05）。ICI182780（1 μmol/L）能部分阻断染料木黄酮（30 μmol/L）对p-AMPK的调节作用（P＜0.05）。结论：染料木黄酮能抑制3T3-L1细胞成脂分化,其机制可能是染料木黄酮一方面下调FAS蛋白表达,抑制脂肪合成,另一方面激活AMPK-HSL途径促进脂肪分解;染料木黄酮还可能通过非雌激素受体途径抑制3T3-L1细胞成脂分化。     

Enrichment of Genistein in Soy Protein Concentrate with Hydrocolloids and β-glucosidase

ABSTRACT: The effects of hydrocolloid (propylene glycol alginate, K-carrageenan, citrus pectin, and xanthan gum) additions in soy protein concentrate (SPC) preparation on genistin and genistein retentions were investigated. Additions of xanthan, alginate, pectin, and carrageenan in SPC prepared with the acid leach method gave 0.711, 0.760, 0.792, and 0.825 mg/g genistein, respectively, whereas control SPC prepared without hydrocolloid gave 0.721 mg/g genistein. SPC prepared under optimum conditions for (β-glucosidase hydrolytic activity with xanthan, alginate, carrageenan, and pectin had 0.943, 0.975, 1.015, and 1.132 mg/g genistein, respectively, compared to genistein in control SPC (0.845 mg/g) under the optimum conditions. Combined (β-glucosidase and pectin treatment in SPC preparation resulted in high genistein SPC (1.551 mg/g).     

大豆异黄酮生理功能的研究进展

大豆异黄酮因具有明显的生物学活性已越来越引起社会和学术界的普遍关注,是近年来科学家研究的热点。研究表明,大豆异黄酮与人类健康密切相关,具有许多生理功能,如抗肿瘤作用、对血管的防护作用、抗氧化活性、类似女性雌激素作用以及抗激素作用等。文中阐述了大豆异黄酮生理功能的研究进展。     

The bone‐protective effect of a Taiwanese yam (Dioscorea alata L. cv. Tainung No. 2) in ovariectomised female BALB/C mice

BACKGROUND: Yam products have been marketed for treating postmenopausal syndromes. This study investigated the effects of Dioscorea alata L. cv. Tainung No. 2 (TNG yam) on the bone density of ovariectomised (OVX) female BALB/c mice and the mechanism whereby TNG yam exerted this effect. Sham and OVX control groups were fed a control diet while remaining OVX mice were randomly allocated into experimental diets, i.e. yam (630 g TNG powder kg^?1), E2 (20 mg 17β‐oestradiol kg^?1), or genistein (2 g genistein kg^?1) diet. After 12 weeks of feeding, the uterine weight, indices of bone mass and caecal short chain fatty acids were determined. RESULTS: Neither a yam nor genistein diet restored the OVX‐induced uterine atrophy as did the E2 diet. The femoral and lumbar bone mineral density (BMD) of mice fed the yam diet was greater than those of the sham group, respectively (P < 0.05 vs OVX control), while the lumbar BMD of yam and sham groups were similar (P > 0.05 vs sham). The femoral ash and calcium content in the yam group was significantly greater than that in the OVX control group, respectively (P < 0.05 vs OVX control). The total short chain fatty acid content in the caecum, only enhanced in the yam group, was not correlated with the calcium content of either bone or the plasma calcium level. CONCLUSION: TNG yam prevented loss of BMD and improved bone calcium status without stimulating uterine hypertrophy in OVX BALB/c mice. TNG yam may be beneficial for postmenopausal women for preventing bone loss. Copyright © 2008 Society of Chemical Industry     

大豆分离蛋白与染料木素共价交联对蛋白表征和结构的影响

探究大豆分离蛋白和染料木素的共价交联对蛋白表征和结构的影响。制备大豆分离蛋白与不同质量浓度染料木素（0、1.2、1.5、2.0 mg/mL）的共价复合物,通过探究粒径、Zeta电位、浊度、表面疏水性分析蛋白体系的表征变化,并采用紫外分光光度计、荧光分光光度计、傅里叶变换红外光谱仪分析蛋白体系的结构变化。结果表明：大豆分离蛋白与染料木素共价复合后,蛋白的中位径由135.6 μm最低减小至98.0 μm,Zeta电位绝对值由15.0 mV最高增大至21.4 mV,表面疏水性由216.0最低减小至115.5,总巯基含量由31.5 μmol/g最低减小至20.4 μmol/g。与对照组相比,共价复合物的浊度增加,并且实验组中SPI-Ge-1.2组低于SPI-Ge-1.5组和SPI-Ge-2.0组。光谱分析表明染料木素对大豆分离蛋白有猝灭效果,二者共价交联后蛋白质的色氨酸与酪氨酸残基所处的微环境疏水性减少,蛋白质二级结构中α-螺旋含量增多、β-折叠含量减少、β-转角含量增多、无规卷曲含量减少,并且加入1.2 mg/mL的染料木素对大豆分离蛋白的表征特征和结构影响效果更好。本研究结果表明在大豆分离蛋白中加入染料木素后,二者的共价交联能够影响蛋白的表征与结构。     

染料木黄酮7,4′-二-O-β-D-吡喃乳糖苷的合成及表征

为了改善染料木黄酮在机体内作用的靶向性,研究了用具有与肿瘤细胞表面受体定向结合的乳糖来修饰染料木黄酮。采用相转移催化法首次合成标题化合物,并对产物进行了IR、MS、1HNMR和13CNMR结构表征。     

Genistein Inhibits Osteoclastic Differentiation of RAW 264.7 Cells via Regulation of ROS Production and Scavenging

Genistein, a phytoestrogen, has been demonstrated to have a bone-sparing and antiresorptive effect. Genistein can inhibit the osteoclast formation of receptor activator of nuclear factor-κB ligand (RANKL)-induced RAW 264.7 cells by preventing the translocation of nuclear factor-κB (NF-κB), a redox-sensitive factor, to the nucleus. Therefore, the suppressive effect of genistein on the reactive oxygen species (ROS) level during osteoclast differentiation and the mechanism associated with the control of ROS levels by genistein were investigated. The cellular antioxidant capacity and inhibitory effect of genistein were confirmed. The translation and activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 1 (Nox1), as well as the disruption of the mitochondrial electron transport chain system were obviously suppressed by genistein in a dose-dependent manner. The induction of phase II antioxidant enzymes, such as superoxide dismutase 1 (SOD1) and heme oxygenase-1 (HO-1), was enhanced by genistein. In addition, the translational induction of nuclear factor erythroid 2-related factor 2 (Nrf2) was notably increased by genistein. These results provide that the inhibitory effects of genistein on RANKL-stimulated osteoclast differentiation is likely to be attributed to the control of ROS generation through suppressing the translation and activation of Nox1 and the disruption of the mitochondrial electron transport chain system, as well as ROS scavenging through the Nrf2-mediated induction of phase II antioxidant enzymes, such as SOD1 and HO-1.     

Genistein Inhibition of Topoisomerase II�� Expression Participated by Sp1 and Sp3 in HeLa Cell

Genistein (4′, 5, 7-trihydroxyisoflavone) is an isoflavone compound obtained from plants that has potential applications in cancer therapy. However, the molecular mechanism of the action of genistein on cancer cell apoptosis is not well known. In this study, we investigated the effect of genistein on topoisomerase II-α (Topo IIα), an important protein involved in the processes of DNA replication and cell proliferation. The results revealed that inhibition of Topo IIα expression through the regulation of Specificity protein 1 and Specificity protein 3 may be one of the reasons for genistein’s induction of HeLa cell apoptosis.