[show abstract][hide abstract] ABSTRACT: (-)-Epigallocatechin-3-gallate (EGCG), the most abundant catechin found in green tea, effectively reduces body weight and tissue and blood lipid accumulation. To explore the mechanism by which EGCG inhibits cellular lipid accumulation in free fatty acid (FFA) induced HepG2 cell culture, we investigated the proteome change of FFA-induced HepG2 cells exposed to EGCG using two-dimensional gel electrophoresis and mass spectrometry.
In this study, 36 protein spots showed a significant change in intensity by more than 1.5-fold from the control group to the FFA group and from the FFA group to the FFA + EGCG group. Among them, 24 spots were excised from gels and identified by LC-MS/MS. In total, 18 proteins were successfully identified. All identified proteins were involved in lipid metabolism, glycometabolism, antioxidant defense, respiration, cytoskeleton organization, signal transduction, DNA repair, mRNA processing, iron storage, or were chaperone proteins. This indicated that these physiological processes may play roles in the mechanism of inhibition of lipid accumulation by EGCG in FFA-induced HepG2 cells. Western blotting analysis was used to verify the expression levels of differentially expressed proteins, which agree with the proteomic results.
From the proteomic analysis, we hypothesized that EGCG reduced cellular lipid accumulation in FFA-induced HepG2 cells through the activation of AMP-activated protein kinase (AMPK) resulting from the generation of reactive oxygen species (ROS). The induction of ROS may be a result of EGCG regulation of the antioxidant defense system. Activation of AMPK shifted some FFA toward oxidation, away from lipid and triglyceride storage, and suppressed hepatic gluconeogenesis. The findings of this study improve our understanding of the molecular mechanisms of inhibition of lipid accumulation by EGCG in HepG2 cells.
[show abstract][hide abstract] ABSTRACT: Renal accumulation of reactive carbonyl compounds (RCCs) has been linked to the progression of diabetic nephropathy. We previously demonstrated that carbonyl stress induces the formation of amino-carbonyl cross-links and sharply increases the content of β-sheet-rich structures, which is the seed of insoluble aggregates formation, and tea catechin (-)-epigallocatechin 3-gallate (EGCG) can reverse this process in vitro and in vivo. In this study, methylated derivative (-)-epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG3"Me) was hypothesized to neutralize carbonyl stress mediating the formation of insoluble ubiquitinated protein (IUP) aggregates, and reduce the early development of diabetic nephropathy.
Diabetes was induced in mice by intraperitoneally injecting alloxan monohydrate (200 mg/kg/d) twice and administering EGCG3"Me by gavage for 15 d. Reagent case and western blot results showed that, in diabetic kidneys, the carbonyl proteins in the serum increased; and in insoluble protein fraction, 4-hydroxynonenal-modified proteins, IUP aggregates and p62 accumulated; FT-IR study demonstrated that the lipid content, anti-parallel β-sheet structure and aggregates increased. EGCG3"Me treatment could effectively reverse this process, even better than the negative control treatment.
EGCG3"Me exhibiting anti-β-sheet-rich IUP aggregate properties, maybe represents a new strategy to impede the progression of diabetic nephropathy and other diabetic complications.
PLoS ONE 01/2013; 8(9):e75687. · 3.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: BACKGROUND: Fuzhuan brick tea is a kind of microbial fermented tea, which has received increasing attention in recent years owing to its benefits for human health. In this study, the anti-obesity and hypolipidemic effects of Fuzhuan brick tea water extracts (FTEs) were investigated. RESULTS: FTEs consisted of 204.07 ± 3.38 mg g(-1) polyphenol, 109.20 ± 1.36 mg g(-1) flavonoids, and others. The FTEs significantly suppressed the increase of body weight and accumulation of adipose tissue, and reduced the level of serum triacylglycerol, total cholesterol and low-density lipoprotein (LDL) cholesterol in obese rats fed a high-fat diet. Moreover, FTEs attenuated the gene expression of sterol regulatory element binding protein-1c, fatty acid synthase and CCAAT/enhancer binding protein α, which is related to lipogenic metabolism. In contrast, the gene expressions of enzymes involved in energy expenditure and lipodieresis including hepatic peroxisome proliferator-activated receptor α, carnitine palmitoyltransferase 1a and LDL receptor gene expression were increased by FTE treatment. CONCLUSIONS: Our study demonstrates that FTEs have anti-obesity and hypolipidemic functions, suggesting that it might be effective for treatment of obesity and hyperlipemia.
Journal of the Science of Food and Agriculture 08/2012; · 1.76 Impact Factor
[show abstract][hide abstract] ABSTRACT: It is still a major challenge to simultaneously isolate artemisinin and its precursors, especially dihydroartemisinic acid and artemisinic acid, from herbal Artemisia annua. A rapid, economical and automatical chromatographic separation process to isolate and purify artemisinin, dihydroartemisinic acid and artemisinic acid at the same time on a preparative scale was developed. The procedure included solvent extraction of ground Artemisia annua leaves by refluxing and purification of crude extract by preparative reverse-phase high-performance liquid chromatography (RP-HPLC). Fractions containing artemisinin and its precursors were collected and identified by gas chromatography and mass spectrometry. High purity of artemisinin, dihydroartemisinic acid and artemisinic acid was obtained by preparative HPLC with a C(18) column and 60% acetonitrile in water as the mobile phase. The techniques described here are useful tools for the preparative-scale isolation of artemisinin and its precursors in a fast, cost-effective and environmental friendly manner.
[show abstract][hide abstract] ABSTRACT: White leaf No.1 is a typical albino tea cultivar grown in China and it has received increased attention in recent years due to the fact that white leaves containing a high level of amino acids, which are very important components affecting the quality of tea drink. According to the color of its leaves, the development of this tea cultivar is divided into three stages: the pre-albinistic stage, the albinistic stage and the regreening stage. To understand the intricate mechanism of periodic albinism, a comparative proteomic approach based on two-dimensional electrophoresis (2-DE) and mass spectrometry was adopted first time to identify proteins that changed in abundance during the three developmental periods.
The 2-DE results showed that the expression level of 61 protein spots varied markedly during the three developmental stages. To analyze the functions of the significantly differentially expressed protein spots, 30 spots were excised from gels and analyzed by matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry. Of these, 26 spots were successfully identified. All identified protein spots were involved in metabolism of carbon, nitrogen and sulfur, photosynthesis, protein processing, stress defense and RNA processing, indicating these physiological processes may play crucial roles in the periodic albinism. Quantitative real-time RT-PCR analysis was used to assess the transcriptional level of differentially expressed proteins. In addition, the ultrastructural studies revealed that the etioplast-chloroplast transition in the leaf cell of White leaf No. 1 was inhibited and the grana in the chloroplast was destroyed at the albinistic stage.
In this work, the proteomic analysis revealed that some proteins may have important roles in the molecular events involved in periodic albinism of White leaf No. 1 and identificated many attractive candidates for further investigation. In addition, the ultrastructural studies revealed that the change in leaf color of White leaf No. 1 might be a consequence of suppression of the etioplast-chloroplast transition and damage to grana in the chloroplast induced by temperature. These results provide much useful information to improve our understanding of the mechanism of albinism in the albino tea cultivar.
[show abstract][hide abstract] ABSTRACT: As tea is one of the most popular beverages consumed worldwide, it is important for customers and business investigators to develop an easy and reliable method to discriminate between different types of teas from each other. A total of eighty-seven types of various white, green, oolong, black and Puer teas were collected from the major tea estates in China, and their catechin contents and volatile compounds were compared by high performance liquid chromatograph and gas chromatograph mass spectrometer. It was found green tea contained the highest concentrations of total catechins, (−)-epicatechin gallate (ECG) and (−)-epigallocatechin gallate (EGCG), while oolong teas contained the highest concentrations of (−)-epigallocatechin (EGC) among these five types of teas. The aroma composition and their quantities in different types of teas varied quite widely. The concentration of ECG, EGCG, pentanal, hexanal, methyl jasmonate, indole, (E,E)-2,4-hexadienal and 1,2,3-trimethoxybenzene was shown to be different and could be used to discriminate white, green, oolong, black and Puer teas. The result showed that different types of teas could be partially classified by cluster analysis using index of individual catechins and volatile components.
International Journal of Food Science & Technology 04/2011; 46(7):1406 - 1412. · 1.24 Impact Factor
[show abstract][hide abstract] ABSTRACT: Emerging evidence supports health-promoting properties of post-fermented Chinese Brick Tea. Fu Zhuan Tea, fermented with the fungus, Erotium cristatum, contains a unique phytochemical profile attributed to its unique method of processing. Fu Zhuan Tea has been shown to activate pancreatic enzymes and regulate blood lipids in laboratory models. Regulation of blood lipid levels by Fu Zhuan Tea consumption was examined in an observational pilot study of volunteers with elevated LDL cholesterol that were not taking any prescription lipid lowering medications. Significant changes in blood lipids were detected after 120 days of daily consumption. Fu Zhuan Tea fractionation led to the investigation of six compounds for regulation of transcription factors involved in lipid metabolism, including Farnesoid X receptor (FXR), Liver X-activated Receptor (LXR) and two isoforms of the Peroxisome Proliferator-Activated Receptors (PPARγ and PPARδ). Reporter gene assays with liver cells revealed dose dependent differences in regulation of transcription factor activation. These data reveal potential therapeutic efficacy and candidate molecular targets for Fu Zhuan Tea, and provide rationale for chemical characterization of bioactive fractions and investigation of therapeutic efficacy in cardiovascular disease and type 2 diabetes.
Food Research International - FOOD RES INT. 01/2011; 44(9).
[show abstract][hide abstract] ABSTRACT: In order to estimate landmark direction angle to localize robot in an artificial environment based on omnidirectional image, this paper introduces the method about low pass filter noise elimination, artificial landmark features detection and landmark direction angle estimation algorithms. Landmark feature detection proposed that red landmark pixels beyond the threshold were extracted as a small area and the center of gravity was calculated for the extracted small area representing the candidate of one landmark. The blue patch provides as compensation to further distinguish the landmark from other objects in a complex environment. Two type experiments were done. The landmark detection experiment proved that the low-pass filter noise elimination was effective. Another experiment was conducted on the level cement ground in a 5×5-m area in the sunlight. 16 test positions were chose to test. Results showed that the MAE and RMSE of landmark direction angles were 1.12 degree and 1.30 degree, respectively. The proposed direction angle estimation method is high accuracy, low time-consuming computation and good robustness.
Information and Automation (ICIA), 2010 IEEE International Conference on; 07/2010
[show abstract][hide abstract] ABSTRACT: Properties of leaf and infusion colours, chemical components and volatile flavour compounds of oolong teas and their correlation with perceived quality score given by tea-tasting panel were analysed. The scores for appearance and infused leaf correlated strongly with concentrations of chlorophyll a (chl a), chlorophyll (chl b) and chlorophyll (chl) (r = 0.355–0.433, P < 0.05) and the total quality score (TQS) positively correlated with concentrations of chl a, chl b and chl (r = 0.517–0.533, P < 0.01). The perceived taste score and TQS positively correlated with the concentration of total free amino acid (r = 0.514, 0.694, P < 0.01) and theanine (r = 0.500, 0.684, P < 0.01). The volatile composition and their quantities varied widely among oolong tea samples. Nerolidol, indole, benzeneacetaldehyde, linalool, linalool oxide I, hexanal, benzyl nitrile, geraniol and 1-penten-3-ol were prevailing volatile compounds detected in most of oolong tea samples. Principal component analysis screened ten principal components with the first three (glutamic acid, total catechins and benzeneacetaldehyde) explaining 27.86%, 20.00% and 14.46% of the total variance, respectively. Regression analysis upon the ten principal components formulated a prediction model on the total quality score with 78.5% probability. The result showed that oolong teas could be partially classified by cluster analysis based on principal components.
International Journal of Food Science & Technology 04/2010; 45(5):913 - 920. · 1.24 Impact Factor
[show abstract][hide abstract] ABSTRACT: Malaria is a global health problem that threatens 300-500 million people and kills more than one million people annually. Artemisinin is highly effective against multidrug-resistant Plasmodium falciparum and it has been widely used as part of the artemisinin-based combination therapies against malaria. To elucidate the biosynthetic pathway of artemisinin and to clone related genes in Artemisia annua, differentially expressed genes between blooming flowers and flower buds were isolated and characterized by a combined approach of suppression subtractive hybridization (SSH) and metabolite analysis. A total of 350 cDNA clones from a subtractive cDNA library were randomly picked, sequenced and analyzed and 253 high-quality sequences were obtained. BLASTX comparisons indicated that about 9.9 % of the clones encoded enzymes involved in isoprenoid (including artemisinin) biosynthesis. The expression of 4 gene transcripts involved in artemisinin biosynthesis was examined by RT-PCR and the results confirmed the higher expression of these transcripts in blooming flowers than in flower buds. In addition, 2 putative transcript factors transparenta testa glabra 1 (TTG1) and ENHANCER OF GLABRA3 (GL3), which promote trichome initiation, were presented in the library. Finally, this study demonstrated that the increase of expression level of the putative TTG1 gene correlated with the improvement of glandular trichome density and artemisinin production in A. annua leaves. The subtractive cDNA library described in the present study provides important candidate genes for future research in order to increase the artemisinin content in A. annua.
[show abstract][hide abstract] ABSTRACT: Malaria threatens 300-500 million people and kills more than one million people annually. Artemisinin has been widely used as part of the artemisinin-based combination therapies against malaria. However, its supply is seriously short due to very small amounts of production of artemisinin in Artemisia annua. Molecular biologic researches aimed at increasing the artemisinin yield in plant have received more and more attention and therefore corresponding quantification methods for artemisinin analysis are urgently needed. A variety of methods for determination of artemisinin have been developed but they cannot be applied when only very little plant material is available or the material should be kept live, which often occurs in molecular biologic researches. The present work developed a simple, fast and low toxic micro-scale analysis procedure for determination of artemisinin in a single leaf or flower of living Artemisia annua using improved gas chromatography with electron-capture detection. The recovery of >95% was achieved by vortex of a piece of fresh leaf in 1 mL ethyl acetate for 2 min at room temperature. This method provides a powerful tool for biosynthesis study of artemisnin, high-throughput screening high-yield clone in an early stage, or real-time quality control of Artemisia annua crop.
[show abstract][hide abstract] ABSTRACT: Artemisinin demand has increased sharply since the World Health Organization recommended its use as part of the artemisinin combination therapies in 2001. The area for the crop cultivation has expanded in Africa and Asia and simpler and affordable methods for artemisinin analysis are needed for crop quality control. This work presented a novel chromatographic method of artemisinin analysis using gas chromatography with electron-capture detection. The sample extraction and preparation involved a single-solvent one-step extraction, with samples being analyzed in the extraction solvent directly after extraction. This method was accurate and reproducible with over 97% recoveries. The limit of detection was less than 3 microg/mL and the limit of quantification was less than 9 microg/mL, allowing samples as low as 100mg dry weight to be analyzed for artemisinin. The method can be applied to quality control of commercial plant extracts and to artemisinin-derived pharmaceuticals.
Journal of Chromatography 06/2008; 1190(1-2):302-6. · 4.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: The present study describes a rapid and sensitive HPLC method for the quantification of huperzine A (HupA) in Huperzia crispata (Huperziaceae). The sample extraction and preparation involved a simple, time-saving, single-solvent extraction, with each sample being analyzed within 12 min. The mobile phase was ammonium acetate (0.1 M, pH 6.0)--methanol (64 + 36, v/v) at a flow rate of 1.0 mL/min. Detection was at 308 nm. The calibration curve was linear from 0.049 to 7.84 microg (R2 = 0.9997), with intraday and interday precision RSD of less than 2%. The extraction recovery rate was over 98.49%. Quantification of HupA was performed using this modified method, and the content of HupA was 1.86 times higher in the whole plant of H. crispata (218.17 +/- 1.55 microg/g) than in that of H. serrata (117.03 +/- 2.97 microg/g). In the whole plant of H. crispata, HupA mainly accumulated in the actively growing shoot tips, the apical bud, and the 10 youngest leaves, reaching 455.23 +/- 2.97 microg/g. The content of HupA in the samples from sunshine-sheltered sites was 3.45 times higher than in that from sunshine-abundant sites. The satisfactory results indicate that this modified method can be applied in the quality control of large-scale Huperziaceae plant extracts and that changes should be made in the cultivation of H. crispata so as to maximize the production of HupA.
Journal of AOAC International 93(5):1428-35. · 1.23 Impact Factor