Phytochemicals and bioactivities of Goji (Lycium barbarum L. and Lycium chinense Mill.) leaves and their potential applications in the food industry: a review

Goji leaves, including Lycium barbarum L. and Lycium chinense Mill. leaves, have been used as traditional Chinese medicine materials for nourishing kidneys and enhancing eyesight but are mostly discarded at present. The nutritional composition of Goji leaves is similar to that of Goji berries even some components are more plentiful in the former, such as flavonoids. This paper firstly reviews the main phytochemicals of Goji leaves. Also, updated information on the biological activities of Goji leaves is narrated. Based on the long use history and existing bioactivity data on Goji leaves, we propose their potential applications in the food industry and encourage further investigation on Goji leaves.     

Enhancing the lipolysis-stimulating activity of soy protein using limited hydrolysis with Flavourzyme and ultrafiltration

The aim of this study was to explore the lipolysis-stimulating activity of soy protein isolate (SPI) hydrolysate using 3T3-L1 adipocytes. Intracellular triglyceride residue (TR) was employed as a marker for lipolysis in cells. The lower TR represents the better lipolysis-stimulating activity. SPI was hydrolysed with Flavourzyme for 2 h to obtain the hydrolysate FH2h, which showed lipolysis-stimulating activity in adipocytes at 400–1600 ppm levels. The sequential fractionation of FH2h with 30–0.3 kDa molecular weight cut-off (MWCO) membranes in order to obtain a 1 kDa retentate resulted in further enhancement of its lipolysis-stimulating activity in the cells. The TR decreased significantly from 2.73 to 2.30 μmole/mg protein at the 400 ppm level (p < 0.05). Based on the western immunoblot and immunostaining analysis, the 1 kDa retentate promotes lipolysis by increasing phosphorylation and translocation of the hormone-sensitive lipase in 3T3-L1 adipocytes.     

中医药防治肥胖症的研究概况

目的：通过网络药理学和分子对接技术,探讨沙棘抗肥胖的活性成分、靶点和作用机制,并验证其体外抗肥胖效果。方法：使用TCMSP平台检索沙棘活性成分与靶点,收集疾病靶点,利用Venny 2.0.2得沙棘靶点与肥胖靶点的交集。通过STRING数据库平台建立药物靶点-疾病靶蛋白相互作用（PPI）网络。使用David数据库对交集靶点进行分析,得到GO富集分析和KEGG通路分析结果。通过Cytoscape 3.9.1软件构建沙棘成分-靶点-信号通路网络图。使用Autodock Dock 1.5.7和Pymol 2.2.0进行沙棘核心靶点与其成分的分子对接,并进行可视化处理。通过体外实验验证沙棘提取物的抗肥胖作用。结果：筛选得到33个沙棘活性成分,2820个疾病靶点和151个交集靶点。主要活性成分包括黄酮类、维生素类、甾醇类等,关键靶点涉及AKT1、TNF、IL6、TP53、VEGFA、CASP3等。KEGG通路富集分析得到恶性肿瘤通路、脂质与动脉粥样硬化通路、AGE-RAGE信号通路等131条信号通路。分子对接结果表明核心靶点与其对应活性成分对接结果良好。体外实验表明沙棘提取物具有抑制3T3-L1小鼠前脂肪细胞增殖的作用。结论：研究显示沙棘具有多成分、多靶点、多通路协同发挥抗肥胖作用,为其临床研究和产品开发提供了参考。

     

素馨花水提物及其主要成分对3T3-L1细胞成脂分化的影响

该研究探讨素馨花水提物（WE）及其化学成分在3T3-L1前脂肪细胞上的降脂作用。通过将3T3-L1细胞诱导为成熟脂肪细胞,油红Ｏ染色定量和检测甘油三酯（TG）含量判断素馨花样品对脂滴的抑制作用;液质联用仪分析WE中化学成分;MTT检测素馨花及其成分对3T3-L1细胞增殖的影响;Western Blot检测AMPK、C/EBPα、FAS、ACC、CPT1、Bax、Bcl-2等蛋白表达。结果发现,WE能剂量依赖性抑制脂滴形成,其中高浓度WE（500 µg/mL）能降低中性脂质含量23.59%,降低TG含量30.20%,在脂肪积累早期作用最显著,并证明其活性成分主要是橄榄苦苷,含量为13.77%。WE及橄榄苦苷能激活AMPK（123.19%和115.98%）和其下游靶点ACC（451.06%和1 050.0%）的磷酸化、下调其下游靶点FAS（81.72%和60.50%）的蛋白表达,减少脂肪形成,提高CPT1（164.84%和292.19%）蛋白的表达,加快脂肪酸氧化;抑制C/EBPα（68.97%和34.57%）的表达,影响3T3-L1细胞分化;上调Bax（154.60%和139.37%）、下调Bcl-2（41.10%和45.62%）蛋白的表达,诱导脂肪细胞凋亡。结果提示,WE能在3T3-L1细胞分化过程早期抑制细胞生长、分化和脂肪合成并促进脂肪酸氧化从而有效抑制脂质积累,机制上可能通过调控AMPK和Bax/Bcl-2通路。     

Enhancing the anti-adipogenic activity of soy protein by limited hydrolysis with Flavourzyme and ultrafiltration

Limited hydrolysis of soy protein isolate (SPI) with Flavourzyme for 2 h to obtain the hydrolysate (FH2h) revealed much higher suppression of glycerol-3-phosphate dehydrogenase (GPDH) activity and relative lipid accumulation (RLA) than intact SPI in 3T3-L1 preadipocytes during differentiation. Lower GPDH activity or RLA indicates higher anti-adipogenic activity. The GPDH significantly decreased from 673 to 477 U/mg protein (p < 0.05). Sequentially fractionating FH2h with 30–1 kDa (kilo-daltons) molecular weight cut-off (MWCO) membranes to obtain the 1 kDa permeate resulted in further reduction of 59% GPDH activity. When comparing the high-performance size-exclusion chromatography (HPSEC) profiles, the most active peptide fraction for the anti-adipogenic activity was primarily composed of small peptides with molecular weight less than 1300 Da. According to the Western immunoblot analysis, 1 kDa permeate inhibits adipogenesis by affecting the expression of peroxisome proliferators-activated receptor γ (PPARγ) and the CCAAT/enhancer binding protein α (C/EBPα) during 3T3-L1 cells differentiation.     

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。     

不同茶多糖对3T3-L1前脂肪细胞分化的抑制作用比较

本实验利用小鼠3T3-L1前脂肪细胞对绿茶多糖（green tea polysaccharides,GTP）、红茶多糖（black tea polysaccharides,BTP）和乌龙茶多糖（oolong tea polysaccharides,OTP）的减肥作用进行评价。使用气相色谱对GTP、BTP、OTP的单糖组成进行分析。采用噻唑蓝染色法测定3?种茶多糖（tea polysaccharides,TPs）对3T3-L1前脂肪细胞增殖活力的影响。使用流式细胞仪检测3?种TPs对3T3-L1前脂肪细胞细胞周期的影响。采用传统“鸡尾酒”法诱导3T3-L1细胞分化成脂后,测吸光度并计算其分化率。采用甘油磷酸氧化酶-过氧化物酶（glycerol phosphate oxidase-peroxidase,GPO-PAP）法测定细胞中甘油三酯（triglyceride,TG）含量的变化。采用实时荧光定量聚合酶链式反应（real-time polymerase chain reaction,RT-PCR）技术测定与脂质代谢相关基因的表达量。结果显示3?种TPs均能显著抑制3T3-L1前脂肪细胞的增殖与分化（P＜0.05）。添加100?μg/mL的TPs能使细胞分化率显著下降至（62.00±6.61）%（GTP）、（82.95±4.25）%（BTP）、（97.24±5.80）%（OTP）。另外,在细胞培养至第5天检测表明,这3 种TPs显著促进G0/G1期细胞数量的累积,细胞比例分别为（68.52±2.28）%（GTP）、（67.11±1.68）%（BTP）、（59.69±1.35）%（OTP）。RT-PCR分析结果显示TPs可以调控相关脂肪细胞因子的表达。在本研究中,在3T3-L1前脂肪细胞中GTP的减肥作用强于BTP和OTP。TPs的加入上调脂联素的表达从而激活腺苷酸活化蛋白激酶信号通路调控相关脂肪细胞因子的表达,并最终抑制TG的合成与3T3-L1前脂肪细胞的分化。     

发酵糙米乙醇提取物改善3T3-L1脂肪细胞糖脂代谢

探讨发酵糙米乙醇提取物(FBREE)对胰岛素抵抗3T3-L1脂肪细胞糖脂代谢的影响。高浓度葡萄糖加胰岛素刺激诱导3T3-L1脂肪细胞胰岛素抵抗模型。不同浓度FBREE处理细胞24 h后检测培养液中葡萄糖,游离脂肪酸(FFA),甘油,甘油三酯(TG),肿瘤坏死因子(TNF-α)和白介素6(IL-6)水平。实时定量PCR检测细胞氧化物酶体增殖激活受体γ(PPARγ),CAATT增强子结合蛋白(C/EBPα),固醇调节元件结合蛋白(SREBP1c),葡萄糖转运蛋白4(GLUT4),脂肪酸结合蛋白(a P2),蛋白酪氨酸磷酸酶1B(PTP1B),葡萄糖转运蛋白(GLUT-4),激素敏感脂肪酶(HSL)和TNF-α,IL-6,单核细胞趋化蛋白-1(MCP-1)和C反应蛋白(CRP)的m RNA表达。FBREE具有促进细胞葡萄糖利用,降低细胞内TG含量,减少FFA及甘油溢出的能力。与模型组细胞相比,高浓度FBREE(100μg/m L)处理的细胞中TG和FFA水平分别下降52.44%和37.83%。FBREE同时能减少模型细胞TNF-α和IL-6的分泌,高浓度FBREE(100μg/m L)分别抑制TNF-α(21.18%),IL-6(31.39%),MCP-1(40.09%)和CRP(41.73%) mRNA表达。此外,FBREE有效降低细胞中PPARγ,C/EBPα,SREBP1c,PTP1B,a P2和HSL的m RNA表达,上调GLUT-4m RNA表达。结果提示,FBREE能有效促3T3-L1脂肪细胞消耗葡萄糖,分解TG,减少FFA和甘油溢出;调控细胞糖、脂代谢来改善胰岛素抵抗,并降低胰岛素抵抗所致炎症水平的升高。     

免疫低下和食物过敏小鼠外周血中细胞因子的变化

建立小鼠免疫低下模型和食物过敏模型,采用ELISA法检测外周血中细胞因子(IL-4、IL-6、IL-10、IL-12、IFN-γ、TGF-β)的变化。结果表明：与空白对照组比较,腹腔注射环磷酰胺所建立免疫低下小鼠胸腺指数和脾脏指数均有显著降低(P≤0.05),且在第3天达到最低;经卵清蛋白致敏小鼠外周血IgE水平极显著升高(P≤0.01)。免疫低下小鼠外周血中IFN-γ(除第3天外)、TGF-β含量(除第3天外)和IFN-γ/IL-4比值均显著降低,而IL-10水平也有下降趋势;除第2天外IL-6水平显著升高,但IL-4水平只在第3天显著性增高,IL-12先是下降随后呈现增多的趋势;食物过敏小鼠的IL-4、IL-6、IL-10和TGF-β水平均有显著性增高,IFN-γ/IL-4比值有显著性下降,而IFN-γ和IL-12没有明显的变化。因此,免疫低下和食物过敏小鼠的Th1/Th2细胞平衡均向Th2细胞偏移,免疫低下对Th1细胞的影响大,食物过敏对Th2细胞的影响更明显。     

大豆油吸附脱色过程氯离子含量变化及对脱臭油中 3-氯丙醇酯和缩水甘油酯的影响

参照土壤中氯离子检测方法,对常见油脂脱色吸附剂中氯离子的超声提取时间进行优化,并对其氯离子含量进行检测。利用常见的吸附剂对大豆油进行吸附脱色,检测吸附脱色前后油脂中氯离子含量,对不同吸附剂脱色后的油脂进行蒸馏脱臭,检测脱臭油中3-氯丙醇酯(3-MCPD酯)和缩水甘油酯(GEs)含量,分析研究不同吸附剂对油脂吸附脱色过程氯离子含量的影响以及对脱臭油中3-MCPD酯和GEs的影响。结果表明:吸附剂中氯离子的最佳超声提取时间为30 min,10个吸附剂样品中氯离子含量为11. 93～187. 881 mg/kg,差异很大;几种吸附剂在油脂吸附脱色过程对氯离子有不同程度的脱除作用,其中脱除效果最好的YS-900活性炭能使油脂中氯离子含量降低83. 21%。对比未进行吸附脱色处理的大豆油,经吸附脱色的大豆油再经脱臭后3-MCPD酯与GEs生成量均有所降低,并且随吸附过程氯离子降低幅度不同,脱臭油中3-MCPD酯与GEs生成量不同:当脱色油中氯离子含量降幅较小时,经脱臭过程GEs生成量明显低于3-MCPD酯;当脱色油中氯离子含量降幅很大时,经脱臭过程3-MCPD酯的生成量低于GEs。以同时降低脱臭过程3-MCPD酯和GEs的生成量为目标,H-2活性炭作为吸附剂是最好的。     

Enhancing the antioxidant effect of α-tocopherol with rosemary in inhibiting catalyzed oxidation caused by Fe and hemoprotein

A mixture of α-tocopherol and rosemary extract (0·035% + 0·035%) was used to inhibit fish lipid oxidation catalyzed by Fe²⁺ or hemoprotein. In a sardine oil model system, a mixture of α-tocopherol and rosemary extract showed a significantly stronger antioxidant effect, as it prolonged the induction period for 10 and 16 days longer than α-tocopherol alone and rosemary extract alone, respectively. In addition, the effective lifetime of α-tocopherol in samples treated with the α-tocopherol and rosemary extract mixture was 10 days longer than in samples with only α-tocopherol added even though the amount of added α-tocopherol in the latter case was twice that for the mixture. In fish dark muscle, the mixture of α-tocopherol and rosemary extract also showed a stronger anti-oxidant effect than either tocopherol or rosemary extract alone. Treatment of samples with this mixture also led to a lower rate of decomposition of highly unsaturated fatty acids, myoglobin and hemoglobin, and triglyceride compared to samples treated with tocopherol or rosemary extract alone.     

Wheat-bug damage in New Zealand wheats: The feeding mechanism of nysius huttoni and its effect on the morphological and physiological development of wheat

Adult Nysius huttoni White were placed in separate cages containing cultivars Rongotea and Karamu at the late anthesis, watery ripe and milky ripe stages of development. Between 84 and 99 % of the matured kernels were injured with the characteristic visible markings of bug-damaged wheat. All the samples contained strong wheat-bug proteinase activity as shown by the incubated SDS-sedimentation test and the disappearance of HMW glutenin subunits from electrophoretograms. Grain infested at late anthesis was most severely affected with shrivelled kernels, high screenings, protein, free amino acids and α-amylase, and low kernel weight, germination capacity and carbohydrate content. Grain infested at the watery ripe and milky ripe stages had values for the above properties closer to uninfested control wheat values. It was suggested that N huttoni sucked sap from lateral sieve tubes in the wheat ovary at late anthesis causing severe disruption to physiological development of the grain. Two commercial lines of wheat showed some characteristics of this effect.