天然植物提取物在果蔬保鲜中应用研究进展

农产品采后保鲜始终是关系到农业可持续发展的重要问题，采用天然植物提取物对果蔬进行保鲜可以减少化学合成杀菌剂对人类健康的不良影响，并有效防止植物病原菌的抗药性。能够应用到果蔬贮藏保鲜中的天然植物提取物主要是天然香辛料和部分中草药, 其提取物对果蔬贮藏保鲜过程中的主要病原菌具有较强的抑制作用。天然防腐保鲜剂的活性成分大多是挥发性精油，主要通过作用于微生物的细胞膜或者能量代谢途径达到抗菌效果。天然植物提取物在果蔬保鲜中的主要应用方式有浸蘸、熏蒸、喷洒或与保鲜纸及涂膜剂等载体相结合，但目前尚缺乏成熟的产业化应用技术。发展植物源果蔬保鲜剂的未来研究应集中于确定植物提取物的活性成分与构效关系，以及产业化应用的系统工程技术。     

Cellular antioxidant activity (CAA) assay for assessing antioxidants, foods, and dietary supplements

A cellular antioxidant activity (CAA) assay for quantifying the antioxidant activity of phytochemicals, food extracts, and dietary supplements has been developed. Dichlorofluorescin is a probe that is trapped within cells and is easily oxidized to fluorescent dichlorofluorescein (DCF). The method measures the ability of compounds to prevent the formation of DCF by 2,2'-azobis(2-amidinopropane) dihydrochloride (ABAP)-generated peroxyl radicals in human hepatocarcinoma HepG2 cells. The decrease in cellular fluorescence when compared to the control cells indicates the antioxidant capacity of the compounds. The antioxidant activities of selected phytochemicals and fruit extracts were evaluated using the CAA assay, and the results were expressed in micromoles of quercetin equivalents per 100 micromol of phytochemical or micromoles of quercetin equivalents per 100 g of fresh fruit. Quercetin had the highest CAA value, followed by kaempferol, epigallocatechin gallate (EGCG), myricetin, and luteolin among the pure compounds tested. Among the selected fruits tested, blueberry had the highest CAA value, followed by cranberry > apple = red grape > green grape. The CAA assay is a more biologically relevant method than the popular chemistry antioxidant activity assays because it accounts for some aspects of uptake, metabolism, and location of antioxidant compounds within cells.     

Role of galloylation and polymerization in cytoprotective effects of polyphenolic fractions against hydrogen peroxide insult

Byproducts and wastes generated by agricultural, food, and forestry industries contain large amounts of polyphenols, which can be potentially used as sources of natural or semisynthetic antioxidants. This study examined and compared the protection against peroxidative damage induced in erythrocytes and 3T3 cell line of polyphenolic fractions from white grape pomace, pine bark, and witch hazel bark. The work pays special attention to the different degrees of polymerization and galloylation of the extracts to contribute to the understanding of their mechanisms of action. Fractions demonstrated different protections against erythrocyte lipid peroxidation, hemolysis, and 3T3 cytotoxicity caused by H(2)O(2). Galloylation is claimed to be related to antioxidant protective capacity, and it is also responsible for the pro-oxidant effect observed at high doses. The results show that not only the percentage of galloylation but also the degree of polymerization are important modulators of their antioxidant capacity. In this sense, it is crucial that novel polyphenolic fractions were prepared attending a value of 3 for the mean degree of polymerization and did not exceed a 30% of galloylation to reach the highest antioxidant capacity with the lowest cytotoxic effects. For this reason, the grape extracts appear to be the best strategy to fight against hydrogen peroxide cell damage.     

Muscadine Grape ( Vitis rotundifolia ) and Wine Phytochemicals Prevented Obesity-Associated Metabolic Complications in C57BL/6J Mice

The objective of this study was to determine the effects of muscadine grape or wine (cv. Noble) phytochemicals on obesity and associated metabolic complications. Muscadine grape or wine phytochemicals were extracted using Amberlite FPX66 resin. Male C57BL/6J mice were given a low-fat diet (LF, 10% kcal fat), high-fat diet (HF, 60% kcal fat), HF + 0.4% muscadine grape phytochemicals (HF+MGP), or HF + 0.4% muscadine wine phytochemicals (HF+MWP) for 15 weeks. At 7 weeks, mice fed HF+MGP had significantly decreased body weights by 12% compared to HF controls. Dietary MGP or MWP supplementation reduced plasma content of free fatty acids, triglycerides, and cholesterol in obese mice. Inflammation was alleviated, and activity of glutathione peroxidase was enhanced. Consumption of MGP or MWP improved insulin sensitivity and glucose control in mice. Thus, consumption of muscadine grape and wine phytochemicals in the diet may help to prevent obesity-related metabolic complications.     

Screening for antioxidant activity in edible plant products: comparison of low-density lipoprotein oxidation assay, DPPH radical scavenging assay, and Folin-Ciocalteu assay

Oxidation of low-density lipoprotein (LDL) has been implicated in atherogenesis. Antioxidants that prevent LDL from oxidizing may reduce atherosclerosis. This study investigated LDL antioxidant activity in edible plant products for development of dietary supplementation to prevent atherosclerosis. Fifty-two kinds of edible plants were extracted using 70% aqueous ethanol solution, and the antioxidant activity of the extracts, which inhibit human LDL oxidation induced by copper ion, was determined on the basis of the oxidation lag time and represented as epigallocatechin 3-gallate equivalent. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and total phenolic content were also measured for comparisons with antioxidant activity in LDL. Plant products showing the greatest activity in LDL oxidation assay were akamegashiwa (Mallotus japonicus) leaf, Japanese privet (Ligustrum japonicum) leaf, green tea [Camellia sinensis (L.) O. Kuntze], and astringent persimmon (Diospyros kaki). The present study revealed high levels of LDL antioxidant activity in plant products for which such activity levels are underestimated in the DPPH radical scavenging assay and Folin-Ciocalteu assay.     

Antioxidant and anti-inflammatory activities of selected medicinal plants containing phenolic and flavonoid compounds

The antioxidant, anti-inflammatory, and cytotoxic activities of water and ethanol extracts of 14 Chinese medicinal plants were investigated and also their total phenolics and flavonoid contents measured. The antioxidant activity was evaluated in a biological assay using Saccharomyces cerevisiae , whereas the radical scavenging activity was measured using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Total phenolics and flavonoid contents were estimated by Folin-Ciocalteu and aluminum chloride methods, respectively. The anti-inflammatory activities of the plant extracts were determined by measuring the inhibition of production of nitric oxide (NO) and TNF-α in LPS and IFN-γ activated RAW 264.7 macrophages. Their cytotoxic activities against macrophages were determined by Alamar Blue assay. Four plants, namely, Scutellaria baicalensis , Taxillus chinensis , Rheum officinale , and Sophora japonica , showed significant antioxidant activity in both yeast model and also free radical scavenging methods. The ethanol extract of S. japonica showed highest levels of phenolics and flavonoids (91.33 GAE mg/g and 151.86 QE mg/g, respectively). A positive linear correlation between antioxidant activity and the total phenolics and flavonoid contents indicates that these compounds are likely to be the main antioxidants contributing to the observed activities. Five plant extracts (S. baicalensis, T. chinensis, S. japonica, Mahonia fortunei , and Sophora flavescens ) exhibited significant anti-inflammatory activity by in vitro inhibition of the production of NO and TNF-α with low IC(50) values. These findings suggest that some of the medicinal herbs studied in this paper are good sources of antioxidants.     

Development of a dual-label time-resolved fluorometric immunoassay for the simultaneous detection of two recombinant proteins in potato

Immunological methods such as ELISA have been traditionally employed to quantify protein levels in plants improved through modern biotechnology. Combined trait products (i.e., plants producing multiple recombinant proteins) created by introducing multiple genetic traits by transformation or traditional breeding methods have prompted the need for the development of analytical assay technologies capable of detecting and quantifying multiple proteins in a single assay. The development of a two-site, sandwich, dual-label, time-resolved fluorometry-based immunoassay (TRFIA) capable of simultaneously quantitating two recombinant proteins (CP4 EPSPS and Cry3A) in plant sample extracts of genetically improved potato cultivars is reported here. The performance characteristics of TRFIA were similar to or exceeded those of current ELISA methods used to detect and quantitate these proteins. TRFIA is a practical and reliable assay for the quantitation of proteins in genetically improved potato plants and offers an alternative approach to conventional ELISA methods with the added benefit of multiple analyte detection.     

不同生长环境下黄顶菊浸提液对多年生黑麦草萌发与生长的影响

为阐明黄顶菊入侵机制,本文在调查不同生境下黄顶菊居群特征的基础上,进一步采用植株浸提液生物测试的方法研究了不同生境、生长密度及替代植被下黄顶菊对多年生黑麦草的化感潜力。结果表明:黄顶菊的化感作用强弱与其入侵能力存在一定相关性,黄顶菊浸提液的化感潜力随浓度升高而增强;不同植株组织浸提液的化感潜力存在显著差异,表现为地下部分明显低于地上部分;不同生境下黄顶菊浸提液的化感潜力顺序为水边>果园>路边,50 g.L 1茎叶浸提液对多年生黑麦草发芽率化感效应敏感指数分别为2.83(水边)、1.31(果园)和0.71(路边);不同密度黄顶菊的茎叶化感潜力顺序为高、中密度区显著大于低密度区,50g.L 1浓度浸提液对多年生黑麦草发芽率化感效应敏感指数分别为3.91(高密度)、4.89(中密度)和0.29(低密度);根部浸提液的化感潜力顺序则相反。不同替代植被下茎叶浸提液的化感效应为混种>单种,而根系浸提液的化感效应敏感指数则差别不大。因此,黄顶菊不同组织器官化感活性物质分配不同,同时其化感效力能够适应环境条件而发生变化,这可能是造成黄顶菊在不同环境中入侵效果差异的原因之一。     

Advantages of soxflo extractions for phytochemical analysis and bioassay screening. 1. Terpenoids

The Soxflo technique was evaluated for the rapid extraction of plant materials (<90 min) at room temperature. Qualitatively similar chromatograms were obtained by gas chromatography and thin layer chromatography (TLC) with Soxflo (SoF) and Soxhlet (SoL) extracts. Sequential solvent extractions by SoF gave slightly higher yields (132%) of five major sesquiterpenoids. TLC revealed that SoF extractions at room temperature were more selective as extracts contained compounds with a narrower range of R(f)() values. This means that the SoF technique offers the potential for one-step extractions and partial fractionation. This study also showed that there were large differences in the volatile composition of dried and fresh Piper fruits: dried fruits had predominantly sesquiterpenoids while fresh fruits had considerable quantities of both mono- and sesquiterpenoids. This is the first report of alpha-guaiene and alpha- and beta-selinene in Piper guineense fruits. It is suggested that the SoF technique can be useful for the screening of large numbers of plants for phytochemicals or for the preparation of plant extracts for subsequent bioassay studies.     

A cell-based screening identifies compounds from the stem of Momordica charantia that overcome insulin resistance and activate AMP-activated protein kinase

Treatment of insulin resistance is a critical strategy in the prevention and management of type 2 diabetes. The crude extracts from all parts of Momordica charantia L. have been reported by many studies for the effective treatment of diabetes and related complications. However, the exact ingredients responsible for the hypoglycemic effect and the underlying mechanism of their actions have not been well characterized because of the lack of a proper assay and screening system. A new cell-based, nonradioactive, and nonfluorescent screening method was demonstrated in this study to screen for natural products from the stem of M. charantia, aiming to identify hypoglycemic components that can overcome cellular insulin resistance. The results suggest triterpenoids being potential hypoglycemic components of the plant and the mechanism underlying their action involving AMP-activated protein kinase.     

Activity-guided discovery of (S)-malic acid 1'-O-β-gentiobioside as an angiotensin I-converting enzyme inhibitor in lettuce (Lactuca sativa)

Angiotensin-converting enzyme (ACE), playing a crucial role in the renin angiotensin aldosterone system, is well-known to catalyze the conversion of the decapeptide angiotensin I into the physiologically active octapeptide angiotensin II, triggering blood pressure increasing mechanisms. To meet the demand for natural phytochemicals as antihypertensive agents in functional food development, extracts prepared from a series of vegetables were screened for their ACE-inhibitory activity by means of a LC-MS/MS-based in vitro assay. By far the highest ACE inhibition was found for a lettuce extract, in which the most active compound was located by means of activity-guided fractionation. LC-MS, NMR spectroscopy, and hydrolysis experiments followed by ion chromatography led to the unequivocal identification of the ACE inhibitor as the previously not reported (S)-malic acid 1'-O-β-gentiobioside. This glycoside represents a novel class of ACE-inhibiting phytochemicals with a low IC(50) value of 27.8 μM. First incubation experiments in saliva and aqueous hydrochloric acid demonstrated the stability of (S)-malic acid 1'-O-β-gentiobioside against salivary glycosidases and stomach acid.     

Databases on food phytochemicals and their health-promoting effects

Considerable information on the chemistry and biological properties of dietary phytochemicals has accumulated over the past three decades. The scattering of the data in tens of thousands publications and the diversity of experimental approaches and reporting formats all make the exploitation of this information very difficult. Some of the data have been collected and stored in electronic databases so that they can be automatically updated and retrieved. These databases will be particularly important in the evaluation of the effects on health of phytochemicals and in facilitating the exploitation of nutrigenomic data. The content of over 50 databases on chemical structures, spectra, metabolic pathways in plants, occurrence and concentrations in foods, metabolism in humans and animals, biological properties, and effects on health or surrogate markers of health is reviewed. Limits of these databases are emphasized, and needs and recommendations for future developments are underscored. More investments in the construction of databases on phytochemicals and their effects on health are clearly needed. They should greatly contribute to the success of future research in this field.     

Evaluation of the antioxidant activity of environmental plants: activity of the leaf extracts from seashore plants

The antioxidant activity of the methanolic extracts of the leaves of 39 plant species was examined. These leaves were collected from the plants growing on subtropical seashores. The activity was evaluated by three kinds of assay methods, which included the DPPH radical scavenging assay, linoleic acid oxidation assay, and oxidative cell death assay. Two extracts from Excoecaria agallocha and Terminalia catappa showed remarkably potent activity in all assay systems. The HPLC analysis of the extracts indicated the presence of the same antioxidant and isolation work for the compound identified ellagic acid. The isolated ellagic acid showed strong antioxidant activity in the assay systems used.     

Chemical composition and biological properties of portuguese wild mushrooms: a comprehensive study

The chemical composition and biological properties of Portuguese wild mushrooms ( Cantharellus cibarius, Hypholoma fasciculare, Lepista nuda, Lycoperdon molle, Lycoperdon perlatum, Ramaria botrytis, Tricholoma acerbum) were evaluated in order to assess these products as sources of nutrients and nutraceuticals. The analyzed mushrooms contain very useful phytochemicals such as phenolics, tocopherols, ascorbic acid, and carotenoids. All of the species proved to have antioxidant activity (measured by four different methods), being more significant for R. botrytis (EC 50 values < 1 mg/mL). Lycoperdon species were resistant to all of the tested microorganisms, and the other samples revealed antimicrobial activity selectively against Gram-positive bacteria, with very low minimal inhibitory concentration, in some cases, even lower than the standard. The combination of bioactive compounds and rich nutritional composition (high contents in protein and carbohydrates, low content in fat with the precious contribution of unsaturated fatty acids and the absence of trans fatty acids) in the mushroom makes it a very special food.     

Galloylated polyphenols as inhibitors of hemoglobin-catalyzed lipid oxidation in fish muscle

The influence of galloyl residues on the antioxidant mechanism of polyphenols to prevent hemoglobin-promoted lipid oxidation was investigated by using polyphenolic fractions with different degrees of galloylation: nongalloylated structures from pine bark (IVP), medium-galloylated from grape pomace (IVG), and high-galloylated from witch hazel bark (IVH). Hemoglobin (Hb) from the pelagic fish horse mackerel (Trachurus trachurus) was employed as a Hb standard. In vitro experiments showed an important increase in the deoxygenation and autoxidation of horse mackerel Hb at acidic pH values. All polyphenolic fractions significantly reduced the redox stability of Hb in buffer solutions, showing a greater deoxygenation and methemoglobin (metHb) formation in the presence of IVH, followed in decreasing order by IVG and IVP. However, galloylated polyphenols (IVH and IVG) were efficient to inhibit the oxidation of the oxygenated Hb (OxyHb) and the formation of lipid oxidation products in chilled washed fish muscle. This antioxidant activity of galloylated proanthocyanidins showed a positive relationship with the phenolic concentration. Polyphenols devoid of galloyl groups (IVP) were less active to prevent either Hb oxidation or lipid oxidation in fish muscle. The results draw attention to the potential role of galloyl residues to lessen Hb-catalyzed lipid oxidation in muscle and to maintain Hb in reduced and oxygenated states, which exhibit lower pro-oxidant activity as compared to the metHb and deoxyHb species.     

Antimycotic activities of selected plant flora,growing wild in Lebanon,against phytopathogenic fungi

Petroleum ether (PE) and methanolic extracts of nine wild plant species were tested in vitro for their antimycotic activity against eight phytopathogenic fungi. The efficacy of PE extracts against all pathogens tested was higher than that of methanolic extracts. Wild marjoram (Origanum syriacum) PE extract showed the highest and widest range of activity. It resulted in complete inhibition of mycelial growth of six of eight fungi tested and also gave nearly complete inhibition of spore germination of the six fungi included in the assay, namely, Botrytis cinerea, Alternaria solani, Penicillium sp., Cladosporium sp., Fusarium oxysporum f. sp. melonis, and Verticillium dahlia. The other plant extracts showed differential activities in the spore germination test, but none was highly active against mycelial growth. Inula viscosa and Mentha longifolia were highly effective (>88%) in spore germination tests against five of six fungi tested, whereas Centaurea pallescens, Cichorium intybus, Eryngium creticum, Salvia fruticosa, and Melia azedarach showed >95% inhibition of spore germination in at least two fungi. Foeniculum vulgare showed the least antimycotic activity. Fractionation followed by autobiography on TLC plates using Cladosporium sp. as a test organism showed that O. syriacum PE extracts contained three inhibition zones, and those of Inula viscosa and Cichorium intybus, two, whereas the PE extracts of the remaining plants showed each one inhibition zone. Some of the major compounds present in these inhibition zones were identified by GC-MS. The possibility for using these extracts, or their mixtures, to control plant diseases is discussed.     

Ergosterol profiles, fatty acid composition, and antioxidant activities of button mushrooms as affected by tissue part and developmental stage

This article investigated the mycochemical profiles and the antioxidant activities of the lipophilic extracts of the white and brown button mushrooms. We found that only free ergosterols were present in both mushrooms at 2.04-4.82 mg/g dry matter (DM). Ergosterol concentration was higher in early growth stages but decreased as the mushrooms grew, and it distributed evenly between the caps and stems during early developmental stages but accumulated more in the caps after maturation. The photochemiluminescence (PCL) values of the two mushrooms were 5.49-10.48 nmol trolox equivalent/mg DM, and the EC50 values of 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay ranged 20.19-41.49 mg DM/μg DPPH. The ergosterol content positively correlated with the antioxidant activities (r2>0.89). The total fatty acid content was 8.7 mg/g DM in the white and 5.1 mg/g DM in the brown button mushroom and contained mainly linoleic, palmitic, and stearic acids. Our data provide guidance for optimized harvesting time of mushrooms and maximized health benefits.     

Effect of yerba mate (Ilex paraguariensis) tea on topoisomerase inhibition and oral carcinoma cell proliferation

Tea flavonoids have antitopoisomerase activity and can inhibit cell proliferation. The objectives of this study were to determine the phenolic content of yerba mate tea products (MT) (Ilex paraguariensis) and evaluate their capacity to inhibit topoisomerase I (Topo I) and II (Topo II) activities and oral carcinoma cell proliferation. Total polyphenols of aqueous extracts of dried MT leaves were measured by the Folin-Ciocalteau assay, using chlorogenic (CH) and gallic (GA) acids as standards. Topoisomerase inhibition was determined by a clone-forming assay, which uses yeast (Saccharomyces cerevisiae) strains as a model. Controls included dimethyl sulfoxide (1.66%); camptothecin (50 microg/mL), a Topo I inhibitor; and amsacrine (100 microg/mL), a Topo II inhibitor. Cytotoxicity studies were conducted using a nontumorigenic human keratinocyte cell line HaCaT and two human squamous cancer cell lines (SCC-61 and OSCC-3). MT was found to be a rich source of phenolic compounds. Total polyphenol content of various commercially available traditional MT products ranged from 236 to 490 mg equiv of CH/g of dry leaves. Such levels were significantly different among products depending on their origin (P < 0.001). Higher anti-topoisomerase II activity was observed against JN394t(2-4) strain for Nobleza Gaucha MT (IC50 = 0.43 microg equiv of CH) in comparison to GA (IC50 = 112 mM) and CH (IC50 > 1500 mM). MT showed catalytic anti-topoisomerase activity against Topo II but not against Topo I. In addititon, MT exhibited dose-dependent cytotoxicity against all squamous cell lines tested. In comparison to premalignant cell line HaCaT [28 microg equiv of (+)-catechin mL(-1)], the cell line SCC-61 [21 microg equiv of (+)-catechin mL(-1)] was the most sensitive to MT, resulting in 50% inhibition of net cell growth. It is concluded that MT is rich in phenolic constituents and can also inhibit oral cancer proliferation. The effect on cancer cell proliferation may be mediated via inhibition of topoisomerase II. The lack of correlation between polyphenol content and the inhibition of topoisomerases suggests that the effect of MT on topoisomerase inhibition may be due to other still unidentified biologically active phytochemicals constituents.     

Extract of lotus leaf ( Nelumbo nucifera ) and its active constituent catechin with insulin secretagogue activity

The effect of lotus leaf ( Nelumbo nucifera Gaertn.) on diabetes is unclear. We hypothesized that lotus leaf can regulate insulin secretion and blood glucose levels. The in vitro and in vivo effects of lotus leaf methanolic extract (NNE) on insulin secretion and hyperglycemia were investigated. NNE increased insulin secretion from β cells (HIT-T15) and human islets. NNE enhanced the intracellular calcium levels in β cells. NNE could also enhance phosphorylation of extracellular signal-regulated protein kinases (ERK)1/2 and protein kinase C (PKC), which could be reversed by a PKC inhibitor. The in vivo studies showed that NNE possesses the ability to regulate blood glucose levels in fasted normal mice and high-fat-diet-induced diabetic mice. Furthermore, the in vitro and in vivo effects of the active constituents of NNE, quercetin, and catechin, on glucose-induced insulin secretion and blood glucose regulation were evaluated. Quercetin did not affect insulin secretion, but catechin significantly and dose-dependently enhanced insulin secretion. Orally administered catechin significantly reversed the glucose intolerance in high-fat-diet-induced diabetic mice. These findings suggest that NNE and its active constituent catechin are useful in the control of hyperglycemia in non-insulin-dependent diabetes mellitus through their action as insulin secretagogues.