白莺山古茶的化学成分分析与栽培茶树的起源

应用高压液相色谱(HPLC)技术,对白莺山古茶的大理茶素、儿茶素类、茶掊素、没食子酸、咖啡因和茶氨酸的含量进行分析.同时,应用分光光度法测定茶多糖和茶多酚含量.在与野生大理茶及栽培大叶茶(普洱生茶)进行多成分比较的基础上,结合形态性状的变异与进化,讨论白莺山古茶种质资源的多样性与野生大理茶和栽培大叶茶的相互关系.分析研究结果不仅为白莺山古茶的品质评价提供可靠的科学数据,为白莺山丰富的古茶种质资源的深人系统研究和合理开发利用提供基础,同时,通过多种过渡类型的化学成分分析与比较,为栽培大叶茶的起源和大理茶作为大叶茶的野生基源之一的假说提供了植物化学方面的证据.     

Antimicrobial Activities of Tocklai Vegetative Tea Clones

Thirty-one Tocklai vegetative (TV) tea clones contained caffeine and total catechin 44.39 and 227.55 mg/g dry weight of leaves, respectively. The (−)-epigallocatechin gallate (EGCG) was the most abundant (109.60 mg/g) followed by -(−)-epigallocatechin (EGC, 44.54 mg/g), (−)-epicatechin gallate (ECG, 41.74 mg/g), (−)-epicatechin (EC, 27.42 mg/g) and +catechin (4.25 mg/g). Total catechins were highest in TV 20 (509.7 mg/g) and lowest in TV 6 (71.7 mg/g). The tea clones that contain high level of total catechin exhibited the strongest antimicrobial activity. Among caffeine and flavanol compounds, theaflavins (TF) present in black tea possess a similar antimicrobial potency as EC present in fresh leaves, and that the conversion of catechins to TF during fermentation in making black tea tends to alter their antimicrobial activities. The bioactive molecules other than catechins present in tea leaves may also contribute towards antimicrobial activity.     

The antioxidant and pro-oxidant activities of green tea polyphenols: A role in cancer prevention

Green tea (Camellia sinensis) is rich in catechins, of which (−)-epigallocatechin-3-gallate (EGCG) is the most abundant. Studies in animal models of carcinogenesis have shown that green tea and EGCG can inhibit tumorigenesis during the initiation, promotion and progression stages. Many potential mechanisms have been proposed including both antioxidant and pro-oxidant effects, but questions remain regarding the relevance of these mechanisms to cancer prevention. In the present review, we will discuss the redox chemistry of the tea catechins and the current literature on the antioxidant and pro-oxidative effects of the green tea polyphenols as they relate to cancer prevention. We report that although the catechins are chemical antioxidants which can quench free radical species and chelate transition metals, there is evidence that some of the effects of these compounds may be related to induction of oxidative stress. Such pro-oxidant effects appear to be responsible for the induction of apoptosis in tumor cells. These pro-oxidant effects may also induce endogenous antioxidant systems in normal tissues that offer protection against carcinogenic insult. This review is meant point out understudied areas and stimulate research on the topic with the hope that insights into the mechanisms of cancer preventive activity of tea polyphenols will result.     

Antioxidant synergism of green tea polyphenols with alpha-tocopherol and L-ascorbic acid in SDS micelles

The synergistic antioxidant effect of polyphenols extracted from green tea, i.e. (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin gallate (EGCG) and gallic acid (GA), with alpha-tocopherol (vitamin E) and l-ascorbic acid (vitamin C) against the peroxidation of linoleic acid has been studied in sodium dodecyl sulfate (SDS) micelles. The peroxidation was initiated thermally by a water-soluble azo initiator 2,2'-azobis(2-amidinopropane) hydrochloride (AAPH), and the reaction kinetics were studied by monitoring the formation of linoleic acid hydroperoxides and consumption of the antioxidants. It was found that the mixture of the green tea polyphenol, vitamin E and vitamin C could act synergistically to protect lipid peroxidation. Kinetic and mechanistic studies on the antioxidation process revealed that this antioxidant synergism was due to the regeneration of vitamin E by the green tea polyphenol and the regeneration of the latter by vitamin C.     

Antimalarial properties of green tea

We show here that a crude extract of green tea as well as two of its main constituents, epigallocatechin-3-gallate (EGCG) and epicatechin gallate (ECG), strongly inhibit Plasmodium falciparum growth in vitro. Both these catechins are found to potentiate the antimalarial effects of artemisinin without interfering with the folate pathway. The importance of these findings and their mechanistic implications are discussed in view of future therapeutic strategies.     

云南“生态茶”的化学成分

从云南西双版纳产大叶茶（Ｃａｍｅｌｌｉａｓｉｎｅｎｓｉｓｖａｒ．ａｓｓａｍｉｃａ）制成的“生态茶”中分离到１１个化合物和１个混晶,经光谱测定,１１个化合物分别鉴定为没食子酸、（＋）儿茶素、（－）表儿茶素、（－）没食子儿茶素、（－）表没食子儿茶素、（－）表儿茶素－３－０没食子酸酯、（－）表没食子儿茶素－３－Ｏ没食子酸酯、茶素、１,４,６－三役食子酰基－β－Ｄ葡萄糖１－Ｏ－没食子酰基－４,６－（－）－六羟基联苯二甲醚基－β－Ｄ－葡萄糖以及咖啡因;混晶为芦丁和山奈酚－３－芸香糖甙的混合物。这是对国产大叶茶的第一次较详细的化学研究。     

一株具有特异AFLP指纹图谱的杜仲古树

杜仲(Eucomm ia ulm oidesO liver)是原产于我国的传统药用植物,虽然目前杜仲被广泛栽培,但野生杜仲已非常稀少,几乎难以见到。作者在利用AFLP分子标记技术研究杜仲的遗传多样性和居群遗传结构时,发现一株采自神农架自然保护区的树龄200多年的雄性杜仲古树(编号为SNJ5)的AFLP指纹图谱与其它582株树龄为10~52年的栽培杜仲的指纹图谱有显著的不同。在总计为191条多态带中,有44条为高频率显性等位基因带(h ighfrequency dom inant allele bands,HFDAB,即583个个体中只有1~3个个体缺乏的带)。SNJ5缺乏23条HFDAB,其中有12条HFDAB是SNJ5单独缺乏的,与其他582个个体差异显著(p<0.05);另外,SNJ5还具有一条特有带(E ACG/M CTG引物组合的217 bp带)。据此推测这一杜仲古树是原始野生植株,是近年来众多学者认为野生杜仲灭绝后发现的非常重要的种质资源,应加以大力保护。     

Caffeine biosynthesis in young leaves of Camellia sinensis: In vitro studies on N-methyltransferase activity involved in the conversion of xanthosine to caffeine

The aim of this study was to investigate the S -adenosylmethionine dependent N -methyltransferase(s) (NMT) associated with the three methylation steps in the caffeine biosynthesis pathway in tea ( Camellia sinensis L.). NMT activity in cell-free preparations from young leaves was purified by anion-exchange and gel-filtration column chromatography. In both systems, a single zone of NMT activity, with broad substrate specificity was detected. The N-3 position of dimethylxanthine and monomethylxanthines was methylated more readily than N-1 while comparatively little substitution occurred at the N-7 locus. When xanthosine was used as a substrate only the N-7 position was methylated. These results indicate that a single NMT may participate in the conversion of xanthosine to caffeine. The apparent M_r of the NMT, estimated by gel filtration chromatography, was 61 000. The substrate specificity of the NMT is compatible with the operation of a xanthosine → 7-methylxanthosine → 7-methylxanthine → theobromine → caffeine pathway as the main biosynthetic route to caffeine in young tea leaves. The data also indicate that the conversion of 7-methylxanthine → paraxanthine → caffeine may function as one of a number of minor pathways that also contribute to the production of caffeine.     

Liquid chromatographic method for the simultaneous determination of caffeine and fourteen caffeine metabolites in urine

An HPLC method has been developed for the separation and the determination of caffeine and its metabolites in urine samples using a one extraction–analysis run and UV detection. The compounds were extracted by liquid–liquid extraction using chloroform–isopropylalcohol (85:15, v/v). Chromatographic separation was accomplished on an ODS analytical column with a mobile phase containing 0.05% acetic acid/methylalcohol (92.5:7.5, v/v). Compounds were monitored at 280 nm. The method was validated for the determination of AFMU, 1X, 1U, 17X and 17U caffeine metabolites required to assess the metabolic activity of the enzymes subject to in vivo caffeine testing. The validated assay was applied to urine samples from ten healthy volunteers. The method was proved to be suitable to assess simultaneously the enzymatic activity of cytochrome P450 CYP1A2 and CYP2A6, as well as N-acetyltransferase and xanthine oxidase.     

Suppression of Helicobacter pylori-induced gastritis by green tea extract in Mongolian gerbils

Since urease of Helicobacter pylori is essential for its colonization, we focused attention on foodstuffs which inhibit the activity of this enzyme. Among plant-derived 77 foodstuff samples tested, some tea and rosemary extracts were found to clearly inhibit H. pylori urease in vitro. In particular, green tea extract (GTE) showed the strongest inhibition of H. pylori urease, with an IC(50) value of 13 microg/ml. Active principles were identified to be catechins, the hydroxyl group of 5(')-position appearing important for urease inhibition. Furthermore, when H. pylori-inoculated Mongolian gerbils were given GTE in drinking water at the concentrations of 500, 1000, and 2000 ppm for 6 weeks, gastritis and the prevalence of H. pylori-infected animals were suppressed in a dose-dependent manner. Since the acquisition by H. pylori of resistance to antibiotics has become a serious problem, tea and tea catechins may be very safe resources to control H. pylori-associated gastroduodenal diseases.     

儿茶素类化合物的抗结核活性研究进展

近年来,耐多药结核病(multidrug-resistant tuberculosis,MDR-TB)的出现及人类免疫缺陷病毒(human immunodeficiency virus,HIV)与结核分枝杆菌合并感染日益增多,使结核病的防治面临更加严峻的挑战,人们急需开发新型抗结核药物及天然低毒的辅助治疗药物。绿茶中的儿茶素类化合物具有多种生物活性,对多种疾病有一定的辅助疗效。多项研究显示,儿茶素类化合物也具有抗结核活性,其机制包括抑制二氢叶酸还原酶活性、影响分枝菌酸及细胞壁的合成、下调富含色氨酸天冬氨酸的膜蛋白(tryptophan-aspartate containing coat protein,TACO)基因表达以抑制结核分枝杆菌的胞内寄生,降低氧化应激水平,下调结核分枝杆菌85B蛋白和宿主肿瘤坏死因子α(tumor necrosis factor α,TNF-α)表达,从而改善炎症水平。有研究显示,喝绿茶可降低结核分枝杆菌感染风险,儿茶素类化合物可辅助治疗结核病并与抗结核药物有协同治疗作用,但目前对其作用机制的研究还不够深入,需进一步探讨。     

Catabolic pathways and biotechnological applications of microbial caffeine degradation

Catabolism of caffeine (1,3,7-trimethylxanthine) in microorganisms commences via two possible mechanisms: demethylation and oxidation. Through the demethylation route, the major metabolite formed in fungi is theophylline (1,3-dimethylxanthine), whereas theobromine (3,7-dimethylxanthine) is the major metabolite in bacteria. In certain bacterial species, caffeine has also been oxidized directly to trimethyl uric acid in a single step. The conversion of caffeine to its metabolites is primarily brought about by N-demethylases (such as caffeine demethylase, theobromine demethylase and heteroxanthinedemethylase), caffeine oxidase and xanthine oxidase that are produced by several caffeine-degrading bacterial species such as Pseudomonas putida and species within the genera Alcaligenes, Rhodococcus and Klebsiella. Development of biodecaffeination techniques using these enzymes or using whole cells offers an attractive alternative to the present existing chemical and physical methods removal of caffeine, which are costly, toxic and non-specific to caffeine. This review mainly focuses on the biochemistry of microbial caffeine degradation, presenting recent advances and the potential biotechnological application of caffeine-degrading enzymes.     

Changes in antibiotic susceptibility in staphylococci habituated to sub-lethal concentrations of tea tree oil (Melaleuca alternifolia)

Aims: To investigate the effect of sub‐lethal challenge with tea tree oil (TTO) on the antibiotic resistance profiles of staphylococci. Methods and Results: Isolates of methicillin‐resistant/‐sensitive Staphylococcus aureus (MRSA and MSSA) and coagulase‐negative staphylococci (CoNS) were habituated to sub‐lethal concentrations of TTO (72 h). Following habituation, the minimum inhibitory concentrations (MIC) of antibiotics and TTO were determined. Habituated MRSA/MSSA cultures had higher (P < 0·05) MIC values than control cultures for the examined antibiotics. Habituated MRSA/MSSA cultures also displayed decreased susceptibility to TTO. Although the MIC of habituated MRSA/MSSA for the examined antibiotics reverted to control values after subsequent culture in the absence of TTO, the increased MIC against TTO were maintained. When compared with control cultures, habituated CoNS cultures had higher (P < 0.05) MIC values against three‐fifths of the antibiotics examined; no changes in TTO MIC were observed. Conclusions: TTO habituation ‘stress‐hardens’ MRSA and MSSA, evidenced by transient decreased antibiotic susceptibility and stable decreased TTO susceptibility. Although TTO habituation did not decrease susceptibility of CoNS to TTO, such cultures showed transient decreased antibiotic susceptibility. Significance and Impact of the Study: Application of TTO at sub‐lethal concentrations may reduce the efficacy of topical antibiotics used with TTO in combination therapies.     

Antimycotic activity of Melaleuca alternifolia essential oil and its major components

AIMS: The aim of this study was to analyse the antimycotic properties of Melaleuca alternifolia essential oil (tea tree oil, TTO) and its principal components and to compare them with the activity of 5-fluorocytosine and amphotericin B. METHODS AND RESULTS: The screening for the antimycotic activity was performed by serial twofold dilutions in Roswell Park Memorial Institute medium with the inclusion of Tween-80 (0.5%). TTO and terpinen-4-olo were the most active compounds. CONCLUSIONS: The majority of the organisms were sensitive to the essential oil, with TTO and terpinen-4-olo being the most active oils showing antifungal activity at minimum inhibitory concentration values lower than other drugs. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides a sample large enough to determine the antifungal properties of TTO and terpinen-4-olo and suggests further studies for a possible therapeutic use.     

Changes in ceramide levels upon catechins-induced apoptosis in LoVo cells

It has been demonstrated that there is difference in the induction of apoptosis in LoVo cells by (-)-epigallocatechin-3-gallate (EGCG), (-)-epigallocatechin (EGC), and (-)-epicatechin (EC). In this study, we explored changes in ceramide levels upon the three catechins-induced apoptosis in LoVo cells. Addition of C2- and C6-ceramide to LoVo cells mimicked EGCG or EGC in leading to apoptotic death. Further measurement of intracellular ceramide content showed that the treatment of LoVo cells with EGCG or EGC resulted in a rapidly transient increase in ceramide content, and then back gradually to base line level, whereas the action of EC was just opposite to that of EGCG or EGC. These results suggest there is difference in the generation of intracellular ceramide by the three catechins and ceramide may take part in the regulation of EGCG- or EGC-induced apoptosis in LoVo cells.