葡萄植株中白藜芦醇及葡萄素含量动态积累规律研究

对葡萄生长期间不同部位中主要活性成分白藜芦醇和葡萄素含量进行了分析,探索了活性成分代谢积累规律.以湖南省境内的主要葡萄品种红地球的主藤、侧藤、支藤、根、叶、叶柄为研究对象,从2010年4月到2011年1月考察了白藜芦醇与葡萄素含量的动态变化,采用超声波辅助甲醇提取法对实验材料进行白藜芦醇和葡萄素提取,以高效液相色谱法进行白藜芦醇和葡萄素含量测定来考察白藜芦醇和葡萄素在葡萄植株中动态变化的规律.结果表明:葡萄植株中不同部位白藜芦醇和葡萄素含量差异显著,主藤＞侧藤＞支藤＞根＞叶片≈叶柄;各部位白藜芦醇和葡萄素在生长期含量变化趋势是先升后降,尤其是葡萄素变化趋势很明显,白藜芦醇变化趋势较小,各部位芪化物含量在9～10月含量处于峰值.     

葡萄果实生长发育过程中白藜芦醇及其糖苷的HPLC测定

利用高效液相色谱(HPLC)梯度洗脱方法,结合C18固相萃取技术,检测了葡萄果实生长发育过程中顺反式白藜芦醇及其糖苷的动态变化。结果显示,适于酿造红葡萄酒的蛇龙珠和酿造白葡萄酒的白羽品种的白藜芦醇及其糖苷含量的变化规律大不相同。葡萄果实生长发育期间,蛇龙珠果实内的白藜芦醇及其糖苷含量较高,而白羽果实中白藜芦醇含量很低,其糖苷含量则持续下降。     

葡萄、葡萄酒的功能性及其开发利用

本文详述了葡萄和葡萄酒中的功能性成分原花青素、白藜芦醇等及其功效,及其与健康的关系。描述了葡萄及葡萄酒在其他方面的开发利用及发展前景。     

野生葡萄及葡萄酒抗氧化活性和抗菌性研究

选取广西、湖南等地野生葡萄,与经典酿酒葡萄比较,研究抗氧化活性和活性物质,同时监测葡萄酒发酵过程中各指标的动态变化,并对不同品种葡萄酒的抗菌性进行研究。结果表明:赤霞珠的酚类含量和抗氧化活性高于野生葡萄和玫瑰香葡萄,但野生葡萄酒的抗菌性能显著优于赤霞珠和玫瑰香葡萄酒。葡萄酒在发酵过程中其抗氧化活性和酚类物质含量均随发酵过程的进行而升高;总抗氧化活性与总酚含量、氧自由基清除能力与原花青素含量成显著正相关,相关系数均大于0.989;总花色苷含量在发酵初期上升,后期下降,葡萄酒颜色变浅。     

抗真菌的有益重组葡萄的开发

法国Moet Hennessy Louis Vuiton公司(MHLV公司)成功地开发出重组葡萄(这种重组葡萄耐真菌且可酿制有益于健康的葡萄酒)。 该公司所开发的是导入了乙烯合成酶基因的重组葡萄。导入该酶基因,目的是增加葡萄酒中的抗氧化物质、白藜芦醇含量。试管实验证明这种物质由于其抗氧化作用不仅能促进健康,而且还可使葡萄抗真菌。     

抗性砧木对赤霞珠葡萄叶片白藜芦醇合成过程及含量的影响

为了明确不同抗性砧木对赤霞珠葡萄叶片白藜芦醇含量及其合成过程中前体物质和相关代谢酶活性的影响,分析不同抗性砧木与白藜芦醇合成的关系,以获得提高接穗品种赤霞珠葡萄叶片白藜芦醇含量的抗性砧木。该研究选择弗卡(Fercal)、5C、140R、3309M、3309C、SO4、抗砧3号(Kangzhen3)、5BB为砧木与赤霞珠(CS)葡萄进行嫁接,以赤霞珠自根苗为对照(CK),采用高效液相色谱技术,测定成熟葡萄叶片白藜芦醇以及合成白藜芦醇前体物质苯丙氨酸、肉桂酸、香豆酸含量,并对合成白藜芦醇相关代谢酶苯丙氨酸解氨酶(PAL)、肉桂酸-4-羟化酶(C4H)、4-香豆-辅酶A连接酶(4CL)、过氧化物酶(POD)以及多酚氧化酶(PPO)的活性进行测定。结果表明:(1)不同砧木均能提高接穗赤霞珠葡萄叶片白藜芦醇含量11%~46%。(2)在各砧穗组合中,CS/140R嫁接苗叶片的白藜芦醇含量最高达到18.24μg/g,其合成白藜芦醇前体物质苯丙氨酸和香豆酸含量也最高,分别达到38.61和1.06μg/g。(3)在各砧穗组合中,白藜芦醇相关代谢酶PAL活性以CS/3309C嫁接苗叶片最高;嫁接苗叶片代谢酶C4H和4CL活性均高于赤霞珠自根苗;POD和PPO活性均以CS/Fercal的接穗赤霞珠叶片最强。研究发现,不同抗性砧木能显著提高赤霞珠葡萄叶片白藜芦醇含量,相关代谢酶C4H活性对葡萄白藜芦醇的合成至关重要,PPO活性与葡萄叶片白藜芦醇合成也密切相关,CS/140R是8个砧穗组合中提高赤霞珠葡萄叶片白藜芦醇含量最具优势的砧穗组合。     

白藜芦醇在几种植物中各部位的分布

采用液相色谱研究白藜芦醇在紫斑牡丹、花生和巨峰葡萄几种经济植物各部位的分布情况。结果表明紫斑牡丹籽和果荚的含量远高于花生和巨峰葡萄。花生白藜芦醇含量最低。紫斑牡丹、花生和巨峰葡萄各部位白藜芦醇分布情况如下：紫斑牡丹叶、紫斑牡丹侧枝、紫斑牡丹茎和紫斑牡丹根中均不含有白藜芦醇,而紫斑牡丹籽和紫斑牡丹果荚中含有白藜芦醇,其中紫斑牡丹籽白藜芦醇含量为0.87‰,紫斑牡丹果荚白藜芦醇含量为0.26‰。花生叶、花生胚乳和花生胚芽中均不含有白藜芦醇,而花生侧枝、花生壳、花生皮、花生茎、花生侧根、花生主根中含有白藜芦醇含有白藜芦醇,其含量为花生侧枝3 mg·kg^-1、花生茎11 mg·kg^-1、花生壳12 mg·kg^-1、花生皮15 mg·kg^-1、花生侧根5 mg·kg^-1、花生主根10 mg·kg^-1。经检测发现葡萄肉,葡萄皮,葡萄籽,葡萄蒂四部分中含有白藜芦醇,其中葡萄肉白藜芦醇含量为0.017‰,葡萄皮白藜芦醇含量为0.028‰,葡萄籽白藜芦醇含量为0.005‰,葡萄蒂和其邻近枝白藜芦醇含量为0.12‰。检测说明这几种植物中花生皮、花生壳、葡萄皮、葡萄籽、葡萄蒂、牡丹籽、牡丹荚具有一定的药用价值和经济价值。     

紫外处理对酿酒葡萄果皮中白藜芦醇及其二聚体合成的影响

本文研究了紫外线辐射对酿酒葡萄品种蛇龙珠果皮中白藜芦醇及其二聚体合成的影响.研究结果表明,紫外线辐射300 s,在35℃下黑暗中贮存24 h后,可明显提高果皮中白藜芦醇及其二聚体含量.紫外线辐射对不同发育期果皮中白藜芦醇及其二聚体含量的影响并不相同,对白藜芦醇含量来说,果实转色期时紫外诱导效果最好;对白藜芦醇二聚体而言,不同时期的紫外诱导效果差异不显著.紫外线辐射均能提高苯丙氨酸裂解酶和过氧化物酶活性,且苯丙氨酸裂解酶活性与果皮中白藜芦醇的形成之间存在相关性,过氧化物酶活性与果皮中自藜芦醇二聚体的形成存在相关性.     

葡萄与葡萄酒中白藜芦醇的研究进展

白藜芦醇是葡萄酒中具有很多保健作用的一种多酚类物质,目前越受到研究人员的重视,本文着重论述了白藜芦醇的保健作用,在葡萄酒中的含量以及影响因素。     

果梗在梅鹿辄干红葡萄酒酿造中的作用

梅鹿辄是酿造葡糖酒的一种优良品种,梅鹿辄干红葡萄酒是一种较为珍贵的葡萄酒类型。在对其进行酿造和制作的过程中需要采用正确的酿造方式和发酵材料,才能够保证葡萄酒的质量。本文主要是对梅鹿辄干红葡萄酒酿造过程中果梗进行研究,从而进一步分析梅鹿辄干红葡萄酒的酿酒工艺优劣,以提高梅鹿辄干红葡萄酒的质量。     

Resveratrol, a red wine constituent, is a mechanism-based inactivator of cytochrome P450 3A4

Resveratrol, a phytoalexin found in red wine, has been shown to possess antioxidant and antimutagenic properties. Incubation of resveratrol with Sf9 insect microsomes containing baculovirus-derived human cytochrome P450 3A4 (CYP3A4) and NADPH-cytochrome P450 reductase showed that resveratrol inactivated CYP3A4 in a time- and NADPH-dependent manner. Resveratrol, erythromycin and troleandomycin inactivated CYP3A4 at a similar rate (as reflected by k(inact)) whereas the binding affinity to CYP3A4 (as reflected by K(I)) was in the order of: troleandomycin > erythromycin > resveratrol. (K(I) and k(inact) for CYP3A4 inactivation by resveratrol, erythromycin and troleandomycin are 20 microM and 0.20 min(-1), 5.3 microM and 0.12 min(-1) and 0.18 microM and 0.15 min(-1), respectively.) Fractionation studies of red wine showed that fractions that did not contain resveratrol inactivated CYP3A4 significantly. In addition, the resveratrol content in red wine used in the study was too low to account for the degree of CYP3A4 inactivation observed after red wine treatment. Inactivation studies using a variety of red wine types showed that the CYP3A4 inactivation did not correlate to their resveratrol content. In summary, data here showed that resveratrol is an effective mechanism-based inactivator of CYP3A4; however, it is not one of the main red wine constituents that are responsible for CYP3A4 inactivation by red wine. Nevertheless, inactivation of CYP3A4 by resveratrol may cause clinically relevant drug interactions with CYP3A4 substrates.     

Resveratrol improves cognitive function in mice by increasing production of insulin-like growth factor-I in the hippocampus

We examined whether resveratrol increases insulin-like growth factor-I (IGF-I) production in the hippocampus by stimulating sensory neurons in the gastrointestinal tract, thereby improving cognitive function in mice. Resveratrol increased calcitonin gene-related peptide (CGRP) release from dorsal root ganglion (DRG) neurons isolated from wild-type (WT) mice. Increases in tissue levels of CGRP, IGF-I, and IGF-I mRNA and immunohistochemical expression of IGF-I were observed in the hippocampus at 3 weeks after oral administration of resveratrol in WT mice. Significant enhancement of angiogenesis and neurogenesis was observed in the dentate gyrus of the hippocampus in these animals (P<.01). Improvement of spatial learning in the Morris water maze was observed in WT mice after administration of resveratrol. None of the effects of resveratrol observed in WT mice were seen after resveratrol administration in CGRP-knockout (CGRP^−/−) mice. Although red wine containing 20 mg/L of resveratrol produced effects similar to those of resveratrol administrationl in WT mice, neither red wine containing 3.1 mg/L of resveratrol nor white wine exhibited such effects in WT mice. Resveratrol was undetectable in the hippocampus of WT mice administered resveratrol and red wine containing 20 mg/L of resveratrol. These observations strongly suggest that resveratrol increases hippocampal IGF-I production via sensory neuron stimulation in the gastrointestinal tract, thereby improving cognitive function in mice.     

Antioxidant effect of red wine polyphenols on red blood cells

The protective effect of red wine polyphenols against hydrogen peroxide (H(2)O(2))-induced oxidation was investigated in normal human erythrocytes (RBCs). RBCs, preincubated with micromolar amounts of wine extract and challenged with H(2)O(2), were analyzed for reactive oxygen species (ROS), hemolysis, methemoglobin production, and lipid peroxidation. All these oxidative modifications were prevented by incubating the RBCs with oak barrel aged red wine extract (SD95) containing 3.5 mM gallic acid equivalent (GAE) of phenolic compounds. The protective effect was less apparent when RBCs were incubated with wines containing lower levels of polyphenols. Furthermore, resveratrol and quercetin, well known red wine antioxidants, showed lower antioxidant properties compared with SD95, indicating that interaction between constituents may bring about effects that are not necessarily properties of the singular components. Our findings demonstrate that the nonalcoholic components of red wine, mainly polyphenols, have potent antioxidant properties, supporting the hypothesis of a beneficial effect of red wine in oxidative stress in human system.     

Red wine triggers cell death and thioredoxin reductase inhibition: Effects beyond resveratrol and SIRT1

Red wine contains antioxidants and is at moderate amounts believed to exert certain positive health effects. Resveratrol is one of the most studied antioxidants in red wine and has been suggested to activate the longevity- and metabolism-associated histone deacetylase SIRT1. Here we show that relatively low concentrations of resveratrol (0.5-3 μM) specifically inhibited neuronal differentiation of neural stem cells in a SIRT1-dependent manner whereas higher concentrations of resveratrol (≥ 10 μM) induced a SIRT1-independent cell death. Surprisingly, using a cell based assay, we found that small amounts of red wine (1-5% v/v) - but not white wine - induced a massive and rapid cell death of various cell types, including neural stem cells and several cancer cell lines. This red wine-induced cell death was ethanol-, SIRT1- and resveratrol-independent but associated with increased oxidative stress and inhibition of thioredoxin reductase (TrxR) activity. The TrxR inhibition correlated with the red color (absorbance at 520 nm) of the wines demonstrating that pigment components of red wine can exert profound cellular effects. Our results unveil important roles for SIRT1 and TrxR in resveratrol and red wine-mediated effects on progenitor and cancer cells, and demonstrate that cellular responses to red wine may be more complex than generally appreciated.     

Differential inhibition of human cytochrome P450 enzymes by epsilon-viniferin,the dimer of resveratrol: comparison with resveratrol and polyphenols from alcoholized beverages

epsilon-Viniferin, a dimer of resveratrol, was isolated in wine at concentration between 0.5 and 5 microM. As resveratrol and polyphenols from red wine were reported to inhibit cytochrome P450 (CYP) activities, this led us to investigate the inhibitory effects of epsilon-viniferin on human CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2E1, CYP3A4 and CYP4A activities. These effects were compared to those of resveratrol and non volatiles compounds from red wine or various Cognac(R) beverages (enriched with oak-polyphenols). Assays were carried out on human liver microsomes and heterologously expressed CYPs. Ethoxyresorufin, coumarin, benzoxyresorufin, chlorzoxazone, testosterone and lauric acid were used as selective substrates for CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2E1, CYP3A4 and CYP4A, respectively. epsilon-viniferin displayed a more potent inhibitory effect than resveratrol for all the CYP activities tested (Ki 0.5 to 20 microM vs. 10 to 100 microM, respectively). This effect was not due to an inhibition of the NADPH reductase. A particularly potent inhibitory effect was shown for CYP1A1, CYP1B1 and CYP2B6 which are involved in bioactivation of numerous carcinogens. epsilon-viniferin was not a mechanism-based inhibitor of human CYPs. It displayed, like resveratrol, mixed-type inhibitions for all the CYP tested, except for CYP2E1 (non-competitive). Comparison of the inhibitory effects exerted on CYP activities by epsilon-viniferin, resveratrol and non volatile components from red wine or various Cognac beverages showed that neither resveratrol, nor epsilon-viniferin is the main CYP inhibitor present in red wine solids.     

Biological effects of resveratrol

Resveratrol (3, 4', 5 trihydroxystilbene) is a naturally occuring phytoalexin produced by some spermatophytes, such as grapevines, in response to injury. Given that it is present in grape berry skins but not in flesh, white wine contains very small amounts of resveratrol, compared to red wine. The concentrations in the form of trans- and cis- isomers of aglycone and glucosides are subjected to numerous variables. In red wine, the concentrations of the trans-isomer, which is the major form, generally ranges between 0.1 and 15 mg/L. As phenolic compound, resveratrol contributes to the antioxidant potential of red wine and thereby may play a role in the prevention of human cardiovascular diseases. Resveratrol has been shown to modulate the metabolism of lipids, and to inhibit the oxidation of low-density lipoproteins and the aggregation of platelets. Moreover, as phytoestrogen, resveratrol may provide cardiovascular protection. This compound also possesses anti-inflammatory and anticancer properties. However, the bioavailability and metabolic pathways must be known before drawing any conclusions on the benefits of dietary resveratrol to health.     

Pro-autophagic polyphenols reduce the acetylation of cytoplasmic proteins

Resveratrol is a polyphenol contained in red wine that has been amply investigated for its beneficial effects on organismal metabolism, in particular in the context of the so-called “French paradox,” i.e., the relatively low incidence of coronary heart disease exhibited by a population with a high dietary intake of cholesterol and saturated fats. At least part of the beneficial effect of resveratrol on human health stems from its capacity to promote autophagy by activating the NAD-dependent deacetylase sirtuin 1. However, the concentration of resveratrol found in red wine is excessively low to account alone for the French paradox. Here, we investigated the possibility that other mono- and polyphenols contained in red wine might induce autophagy while affecting the acetylation levels of cellular proteins. Phenolic compounds found in red wine, including anthocyanins (oenin), stilbenoids (piceatannol), monophenols (caffeic acid, gallic acid) glucosides (delphinidin, kuronamin, peonidin) and flavonoids (catechin, epicatechin, quercetin, myricetin), were all capable of stimulating autophagy, although with dissimilar potencies. Importantly, a robust negative correlation could be established between autophagy induction and the acetylation levels of cytoplasmic proteins, as determined by a novel immunofluorescence staining protocol that allows for the exclusion of nuclear components from the analysis. Inhibition of sirtuin 1 by both pharmacological and genetic means abolished protein deacetylation and autophagy as stimulated by resveratrol, but not by piceatannol, indicating that these compounds act through distinct molecular pathways. In support of this notion, resveratrol and piceatannol synergized in inducing autophagy as well as in promoting cytoplasmic protein deacetylation. Our results highlight a cause-effect relationship between the deacetylation of cytoplasmic proteins and autophagy induction by red wine components.     

Resveratrol counteracts gallic acid-induced down-regulation of gap-junction intercellular communication

Since reactive oxygen species (ROS) play a key role in carcinogenesis, many studies have focused on the chemopreventive activities of naturally occurring antioxidants. However, the possibility that different antioxidants in food exert opposing effects on carcinogenesis has not been adequately investigated. Gap-junction intercellular communication (GJIC), which is strongly related to carcinogenesis (particularly the tumor promotion stage), may be a suitable model for investigating the tumor-promoting and antitumor-promoting effects of phytochemicals. The present study investigated the possible combined effects of resveratrol and gallic acid (GA), which are major antioxidants in red wine, on GJIC in WB-F344 rat liver epithelial (RLE) cells. GA at 100 microM, but not resveratrol, inhibited GJIC and generated hydrogen peroxide. The GA-induced inhibition of GJIC was recovered by resveratrol, but only partially recovered by catalase. Resveratrol did not attenuate GA-induced generation of hydrogen peroxide, but it did block GA-induced phosphorylation of connexin 43 (Cx43), a key modulator of GJIC. Furthermore, resveratrol down-regulated GA-induced phosphorylation of extracellular signal-regulated kinase (ERK)1/2, one of the critical regulators of Cx43. However, catalase partially blocked the GA-induced phosphorylation of Cx43 and ERK1/2. Collectively, these findings suggest that the combined effects of red wine phenolic phytochemicals on GJIC and antioxidants differ in ROS-mediated carcinogenesis depending on their dosages and structures.