甲状腺素对HepG2细胞氧化损伤的保护作用

研究甲状腺激素(T3)对人肝癌细胞(HepG2)氧化损伤的保护作用。体外建立HepG2细胞H2O2氧化损伤模型,添加不同浓度T3进行保护,测定T3对细胞ROS、存活率、抗氧化酶及相关基因的影响。10-9、10-7、10-5mol/L的T3预处理24 h可分别有效降低50、100、200μmol/L H2O2处理细胞的ROS水平,提高细胞存活率(p0.05)。但10-5mol/L的T3反而会增加50μmol/L H2O2处理细胞的ROS水平,降低存活率(p0.05)。适量T3可显著降低H2O2氧化损伤的HepG2细胞MDA含量(p0.05),显著提高T-AOC、GSH-Px以及SOD活性(p0.05)。但过量T3会增加细胞MDA水平,降低酶活。基因表达也有类似趋势,T3预处理使损伤细胞内PI3K、Nrf2的表达显著提高(p0.05),且与T3作用剂量相关。可见T3对H2O2氧化损伤的HepG2细胞的氧化还原状态具有明显的改善作用,其抗氧化保护功能与提高抗氧化酶的活性及细胞内抗氧化相关基因的表达有关,且T3作用效果与其剂量及细胞损伤水平相关。     

诸葛菜碱Ⅰ对H_2O_2所致HepG2细胞氧化损伤的保护作用

本文采用柱色谱技术对诸葛菜种子的提取物进行分离纯化,采用波谱技术结合化学方法进行结构鉴定,首次获得新化合物,命名为诸葛菜碱Ⅰ,并通过建立H_2O_2诱导的细胞损伤模型,对诸葛菜碱Ⅰ保护H_2O_2所致Hep G2细胞氧化损伤的作用及其机制进行了研究。于H_2O_2刺激前,加入不同浓度的诸葛菜碱Ⅰ预处理Hep G2细胞12 h,然后以400μmol/L H_2O_2损伤细胞2 h建立模型。MTT法检测细胞存活率,微板法检测细胞培养液中LDH活性和细胞MDA含量及SOD、GSH-PX的活性,Western Blot检测细胞内Nrf 2、HO-1蛋白表达,以评价诸葛菜碱Ⅰ对H_2O_2所致Hep G2细胞氧化损伤的保护作用。结果表明,与模型组相比,诸葛菜碱I给药组(53.5、107、214μmol/L)预处理12 h后,增加了细胞存活率(p0.01),显著降低LDH向细胞外液的释放(p0.01),显著降低细胞内MDA含量(p0.05,p0.01),显著提高细胞内SOD、GSH-PX的活性(p0.05,p0.01),增高了Nrf2蛋白水平。因此,诸葛菜碱Ⅰ对H_2O_2所致Hep G2细胞氧化损伤具有保护作用。     

高核苷酸酵母水解物调控HepG2细胞氧化损伤作用

目的:探究高核苷酸酵母水解物的体外抗氧化活性。方法:采用化学法测定高核苷酸酵母水解物对二苯代苦味酰基自由基(DPPH)和羟自由基的清除能力及总还原力。建立游离脂肪酸(FFA)诱导HepG2细胞氧化应激模型,采用噻唑蓝溴化四唑(MTT)法检测高核苷酸酵母水解物对HepG2细胞的活力影响,荧光探针DCFHDA法检测HepG2细胞内活性氧的变化,商业试剂盒检测细胞内氧化损伤标志物丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)的活力。实时定量PCR检测keap1,Nrf2,HO-1和NQO-1的mRNA表达水平。免疫印迹检测Nrf2活化与转位情况。结果:5～50 mg/mL高核苷酸酵母水解物以质量浓度依赖性方式清除体外活性氧自由基,具有显著抗氧化能力。500～1 000μg/mL高核苷酸酵母水解物可显著降低细胞内丙二醛(MDA)的含量,降低活性氧自由基(ROS)水平,提高细胞中超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)的活性。抗氧化酶基因NQO1,Nrf2和HO-1的mRNA表达水平显著增加,keap1 mRNA表达水平显著降低。高核苷酸酵母水解物能够显著增加HepG2细胞核内Nrf2的蛋白质含量,降低其胞浆中的蛋白质含量,进一步阐明高核苷酸酵母水解物通过激活Nrf2通路减轻FFA诱导的HepG2氧化应激损伤作用。     

牛磺酸和FK506对HepG2细胞胆固醇水平的影响

目的:以总胆固醇(Total cholesterol,TC)、游离胆固醇(Free cholesterol,FC)和胆固醇酯(Cholesterol ester,EC)为评价指标,探讨牛磺酸对FK506诱导肝癌细胞Hep G2细胞胆固醇水平代谢异常的影响。方法:首先建立损伤模型,加终浓度为1、5和10μg/m L的FK506,分别在培养24、48 h后测定细胞存活率以及胞内TC、FC和EC水平,确定FK506处理的终浓度;在此基础上,再添加终浓度为0.1、1、10和20 mmol/L的牛磺酸,分别培养24、48 h,测定细胞内TC、FC和EC水平。结果:培养24、48 h,5和10μg/m L FK506处理都使细胞TC、FC水平明显升高(p0.05),EC水平没有明显变化,但10μg/m L FK506作用48 h时,细胞存活率显著下降到70%,因此确定FK506处理的终浓度为5μg/m L。在FK506(5μg/m L)预处理6 h后,0.1、1、10和20 mmol/L的牛磺酸作用24、48 h都可显著降低FK506诱导的细胞TC、FC水平升高(p0.05),但作用24 h的效果优于48 h,以20 mmol/L作用24 h降胆固醇效果最佳。结论:FK506通过增加FC诱导Hep G2细胞胆固醇水平升高,而牛磺酸通过降低FC改善了细胞高胆固醇水平。     

GABA对高糖诱导氧化损伤的RIN-m5f细胞的保护作用和机制

GABA在胰腺中能够发挥保护作用,但是其具体的作用机制尚不明确。为了确定GABA是否对胰岛β细胞具有抗氧化、抗凋亡以及促进胰岛素分泌的作用,作者采用高糖(44mmol/L)诱导建立细胞氧化损伤模型,通过测定细胞存活率、细胞ROS生成量、细胞抗氧化酶活、丙二醛含量、胰岛素分泌量以及相关基因水平,分析探讨不同浓度的GABA(10、50、100μmol/L)对高糖诱导氧化损伤的RIN-m5f细胞的保护作用。结果表明,GABA可以提高RIN-m5f细胞的存活率,降低ROS生成量,显著促进SOD和GSH-Px的活力,降低MDA的浓度,促进胰岛素分泌,提高Nrf2和Bcl-2的mRNA水平,并降低GSK-3β和Bax的mRNA水平。而且现出的这些作用存在量效关系。GABA的保护作用可能通过抑制GSK-3β,进一步抑制Bax并激活Nrf2和Bcl-2等关键因子从而实现。因此,GABA对RIN-m5f具有抗氧化、抗凋亡以及促进胰岛素分泌的作用。     

芦荟多糖对D-半乳糖致HepG2细胞氧化损伤的保护作用

目的:探讨芦荟多糖对HepG2细胞氧化损伤的保护作用,分析其体外抗氧化能力。方法:以HepG2细胞为研究对象,建立D-半乳糖诱导细胞氧化损伤模型,以不同浓度芦荟多糖进行预保护处理。采用细胞增殖与毒性检测试剂盒测定HepG2细胞存活率,生物化学法检测细胞超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶活力,酶联免疫吸附法测定细胞总抗氧化能力及丙二醛（malondialdehyde,MDA）含量,实时荧光定量PCR检测细胞Keap1、Nrf2、NQO1和HO-1基因表达水平。结果:与D-半乳糖模型组比较,25、50、100 μg/mL的芦荟多糖预保护处理能不同程度地提高HepG2细胞存活率;芦荟多糖能显著增强细胞中抗氧化酶的活性,降低细胞MDA的生成量（P<0.05）,且作用效果与芦荟多糖浓度成正比;细胞中Keap1基因表达显著降低,Nrf2、NQO1、HO-1 mRNA表达显著提高（P<0.05）。结论:芦荟多糖可以减轻HepG2细胞的氧化应激损伤,激活Nrf2表达并抑制其泛素化降解,提高下游NQO1、HO-1的转录水平,增强细胞抗氧化酶系活性并调控细胞氧化还原系统,以达到减轻细胞氧化损伤程度、提高细胞抗氧化应激损伤的效果。     

矢车菊素-3-O-葡萄糖苷对H_2O_2诱导细胞氧化损伤的保护作用

目的:研究矢车菊素-3-O-葡萄糖苷(C3G)对人胚肾HEK-293细胞氧化损伤的保护作用。方法:通过检测细胞活力、活性氧(ROS)水平和丙二醛(MDA)水平考察C3G对氧化损伤细胞的影响,并采用Western blot法和q RTPCR检测Nrf2和Keap1的蛋白表达量和m RNA表达量变化。结果表明:C3G预处理氧化应激细胞后,细胞活力显著高于H_2O_2损伤组(p 0.05),并呈浓度依赖性提高; H_2O_2损伤组的DCF荧光强于对照组,ROS相对含量增加到近2倍,1.25、5、20μmol/L C3G预处理后细胞荧光强度逐渐减弱,ROS相对量呈浓度依赖性降低; C3G预处理后细胞MDA水平呈浓度依赖性降低; 1.25、5、20μmol/L C3G处理后,Nrf2的m RNA表达和蛋白表达量均呈浓度依赖性上调,Keap1蛋白表达量显著低于H_2O_2损伤组(p 0.05),5和20μmol/L C3G对Keap1的m RNA下调作用明显。结论:矢车菊素-3-O-葡萄糖苷对HEK-293细胞有保护作用,可降低细胞ROS和MDA水平,其作用机制可能是通过激活Nrf2/Keap1信号通路以抵抗H_2O_2导致的细胞氧化损伤。     

槲皮素对黄曲霉毒素B_1致大鼠肝细胞毒性的保护作用

黄曲霉毒素B_1(AFB_1)是由黄曲霉菌在特定条件下产生的毒性最强的次生代谢产物,广泛存在于玉米、花生等农产品中。槲皮素(Quercetin,QUE)是一种具有抗氧化等生理作用的黄酮类化合物。本文拟研究槲皮素对AFB_1致大鼠肝细胞毒性的保护作用及其机制。CCK-8实验结果表明,AFB_1对大鼠正常肝细胞(BRL细胞)的IC_(50)为23μmol/L,而QUE在低于15μmol/L时未对BRL细胞造成毒性损伤。利用QUE对细胞进行预处理24 h后,再将AFB_1作用于细胞24 h,通过细胞增殖力、活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽s-转移酶(GST)指标的检测,表明QUE能够增强细胞内SOD和GST酶活性,提高细胞自身的抗氧化性,降低由AFB_1引起的MDA和ROS对细胞的氧化损伤程度,进而保持细胞活力。此外,AFB_1可引起促凋亡基因Bax的mRNA表达(p0.05),显著降低AKT、Nrf2和HO-1的转录水平(p0.05),而QUE预处理能显著提高AKT、Nrf2、HO-1和Bcl-2基因的表达(p0.05),进而发挥抗氧化及抗凋亡的作用。因此,槲皮素对黄曲霉毒素B_1致大鼠肝细胞的毒性损伤具有一定的保护作用。     

仙草多糖对HepG2细胞酒精性损伤的保护作用

为研究仙草多糖对肝细胞酒精性损伤的保护作用及机制,选用HepG2细胞,经乙醇诱导,检测仙草多糖处理后细胞的存活率、活性氧水平和脂滴蓄积水平。体外抗氧化水平检测发现,仙草多糖对超氧阴离子自由基和羟自由基有清除作用。与乙醇处理组HepG2细胞相比,100μg/m L和200μg/m L的仙草多糖预处理组细胞的存活率显著上升;同时,仙草多糖处理显著降低了酒精诱导HepG2细胞产生的活性氧水平;另外,用仙草多糖处理酒精诱导的HepG2细胞也显著改善了胞内的脂滴蓄积水平。可见,仙草多糖对HepG2细胞酒精性损伤有保护作用,这作用至少有一部分是因为其具有强抗氧化作用,显著抑制了酒精诱导的活性氧水平的上升。     

基于HepG2细胞评价不同来源虾青素抗氧化能力差异

为探究雨生红球藻源虾青素(Haematococcus pluvialis ester astaxanthin, E-AST)和人工合成虾青素(synthetic astaxanthin, S-AST)抗氧化能力的差异，该研究分析了不同浓度(0、2.5、5、10μmol/L)2种来源虾青素处理24 h后，对HepG2细胞存活率、细胞内不同活性氧分子含量和细胞内抗氧化活性(cellular antioxidant activity, CAA)的影响。结果显示，与对照组相比，E-AST和S-AST对HepG2细胞存活率无明显影响，仅10μmol/L E-AST能够显著提高细胞存活率。荧光探针DCFH-DA检测显示，与对照组比较，除5μmol/L E-AST和2.5μmol/L S-AST外，添加其他浓度的2种虾青素均能够显著降低细胞内活性氧(reactive oxygen species, ROS)含量(P<0.05),但是同一浓度2种虾青素处理组之间不存在显著性差异(P>0.05)。DHR123荧光探针的检测结果则显示，在虾青素处理浓度为5μmol/L时，E-AST对H     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.     

Chlorohydrins of bisphenol A diglycidyl ether (BADGE) and of bisphenol F diglycidyl ether (BFDGE) in canned foods and ready-to-drink coffees from the Japanese market

BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H₂O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.     

European priorities for research to support legislation in the area of food contact materials and articles

A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.     

Analysis of benzo[a]pyrene in spiked fatty foods by second derivative synchronous spectrofluorimetry after microwave-assisted treatment of samples

A simple, rapid and inexpensive method has been developed for the determination of benzo[a     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. II: Certification study

This paper describes the second part of a project undertaken to develop certified mussel reference materials for paralytic shellfish poisoning toxins. In the first part two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin and decarbamoyl-saxitoxin in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the certification exercise. Fifteen laboratories participated in this certification study and were asked to measure saxitoxin and decarbamoyl-saxitoxin in rehydrated lyophilized mussel material and in a saxitoxin-enriched mussel material. The participants were allowed to use a method of their choice but with an extraction procedure to be strictly followed. The study included extra experiments to verify the detection limits for both saxitoxin and decarbamoyl-saxitoxin. Most participants (13 of 15) were able to meet all the criteria set for the certification study. Results for saxitoxin.2HCl yielded a certified mass fraction of <0.07 mg/kg in the rehydrated lyophilized mussels. Results obtained for decarbamoyl-saxitoxin.2HCl yielded a certified mass fraction of 1.59+/-0.20 mg/kg. The results for saxitoxin.2HCl in enriched blank mussel yielded a certified mass fraction of 0.48 +/- 0.06 mg/kg. These certified reference materials for paralytic shellfish poisoning toxins in lyophilized mussel material are the first available for laboratories to test their method for accuracy and performance.     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of the