Transcriptional regulation of bovine elongation of very long chain fatty acids protein 6 in lipid metabolism and adipocyte proliferation.

The elongation of very long chain fatty acids protein 6 (ELOVL6) gene encodes a key enzyme that plays a role in lipogenesis through the catalytic elongation of both saturated and monounsaturated fatty acids. Previous studies have described the high expression of bovine ELOVL6 in adipose tissues. However, transcriptional regulation and the functional role of ELOVL6 in lipid metabolism and adipocyte proliferation remain unexplored. Here, a 1.5 kb fragment of the 5'-untranslated region promoter region of ELOVL6 was amplified from the genomic DNA of Qinchuan cattle and sequenced. The core promoter region was identified through unidirectional 5'-end deletion of the promoter plasmid vector. In silico analysis predicted important transcription factors that were then validated through site-directed mutation and small interfering RNA interference with an electrophoretic mobility shift assay. We found that the binding of KLF6 and PU.1 transcription factors occurred in the region -168/+69. Both perform a vital regulatory function in the transcription of bovine ELOVL6. Overexpression of ELOVL6 significantly upregulated the expression of peroxisome proliferator activated receptor γ (PPARγ), but inhibited the expression of fatty acid-binding protein 4 (FABP4), while silencing of ELOVL6 negatively regulated the messenger RNA expression level of PPARγ, FABP4, ACSL, and FATP1. In addition, ELOVL6 promotes adipocyte proliferation by regulating the cell-cycle genes' expression. Taken together, these findings provide useful information about the transcriptional regulation and functional mechanisms of bovine ELOVL6 in lipid metabolism and adipocyte proliferation in Qinchuan cattle.

The role of electron shuttle enhances Fe(III)-mediated reduction of Cr(VI) by Shewanella oneidensis MR-1.

Chromate is one of the hazardous toxic pollutants. Reduction of Cr(VI) to Cr(III) has shown to reduce the toxicity of chromate. This work examined the reduction of Cr(VI) using an anaerobic batch cultures of Shewanella oneidensis MR-1 containing Fe(III). To do so, 10 mg/L Cr(VI) was reduced to Cr(III) within 3 days along with the oxidization of Fe(II) to Fe(III). The removal rate of Cr(VI) increased with increasing the concentration of Fe(III). In the absence of Cr(VI), the Fe(II) concentration of the batch culture increased with the growth of S. oneidensis MR-1. These data showed that S. oneidensis MR-1 could reduce Fe(III) into Fe(II), resulting in reduction of Cr(VI) to Cr(III). During this process, the anthraquinone-2,6-disulfonate (AQDS) acted as an electron shuttle. Microscopic analysis showed that Cr(VI) had toxic effects on S. oneidensis MR-1 due to the appearance of Cr species on the bacterial surface. Cr2O3 or Cr(OH)3 precipitates formed during Cr(VI) reduction was identified using X-ray photoelectron spectroscopy. The AQDS as an electron shuttle enhanced the Cr(VI) reduction by S. oneidensis MR-1. Microbial reduction of Cr(VI) can be a useful technique for Cr detoxification.

Unexpected Information Demand and Volatility Clustering of Chinese Stock Returns: Evidence from Baidu Index.

This paper employs the Baidu Index as the novel proxy for unexpected information demand and shows that this novel proxy can explain the volatility clustering of Chinese stock returns. Generally speaking, these findings suggest that investors in China could take advantage of the Baidu Index to obtain information and then improve their investment decision.

Coordinate regulation by transcription factors and DNA methylation in the core promoter region of SIRT6 in bovine adipocytes.

Sirtuin6 (SIRT6) is an ADP-ribosyltransferase and NAD+-dependent deacylase of acetyl groups and long-chain fatty acyl groups, and has been shown as a regulator of insulin secretion, glucose metabolism, lipid metabolism, and cancer. In this study, we determined that the bovine SIRT6 showed higher levels of mRNA expression in the testis, longissimus thoracis, and subcutaneous fat tissue. To elucidate the molecular regulation mechanism of bovine SIRT6 expression, we obtained a 2-kb fragment containing the 5'-regulatory region, and the functional proximal minimal promoter of bovine SIRT6 was identified in the -472/-73 bp region. The CCAAT enhancer binding protein beta (CEBPß), paired box 6 (PAX6), Kruppel-like factor 2 (KLF2), myb proto-oncogene protein (CMYB), nuclear respiratory factor 1 (NRF1), and E2F transcription factor 1 (E2F1) binding sites, as transcriptional activators or repressors in the core promoter region of SIRT6, were determined by electrophoretic mobility shift assay (EMSA) experiments and luciferase reporter assays. In addition, the results from methylation assay and luciferase report assay showed that the bovine SIRT6 promoter activity was coordinately regulated by methylation and NRF1 or E2F1 during bovine adipocyte differentiation. Taken together, this study illuminated the underlying mechanism of methylation and transcription regulation of SIRT6 expression in bovine adipocytes.

Homo-Conjugation of Low Molecular Weight Organic Acids Competes with Their Complexation with Cu(II).

Dissolved organic matter (DOM) controls the bioavailability and toxicity of heavy metals in aquatic environments. The observation of decreased conditional binding constants with increasing DOM concentration is not well documented, which may result in significant uncertainties in heavy metal behavior modeling and risk assessment. We used eight low molecular weight organic acids (LMOC) with representative structures to simulate DOM molecules. The interactions between LMOC molecules resulted in the decreased Cu(II)-LMOC binding with increasing LMOC concentrations, but higher pH values than theoretical calculation after mixing LMOC solutions of different pHs. We thus proposed homoconjugation between LMOC molecules through negative charge-assisted H-bond ((-)CAHB). A mathematic model was developed to describe Cu(II)-LMOC complexation ( KC) and LMOC homoconjugation ( KLHL). The increased competition of LMOC homoconjugation over Cu(II)-LMOC complexation, as suggested by the increased ratios of KLHL/ KC, resulted in the apparently decreased Cu(II)-LMOC binding with the increased LMOC concentration. Similar concentration-dependent binding was also observed for DOM. With the identified homoconjugation between DOM molecules, some of the literature data with concentration-dependent behavior may be re-evaluated. This is the first work that quantitatively identified homoconjugation among organic molecules. Both the modeling concepts and results provide useful information in investigating the environmental roles of DOM in mediating metal bioavailability and transport.

TMDB: a literature-curated database for small molecular compounds found from tea.

BACKGROUND: Tea is one of the most consumed beverages worldwide. The healthy effects of tea are attributed to a wealthy of different chemical components from tea. Thousands of studies on the chemical constituents of tea had been reported. However, data from these individual reports have not been collected into a single database. The lack of a curated database of related information limits research in this field, and thus a cohesive database system should necessarily be constructed for data deposit and further application. DESCRIPTION: The Tea Metabolome database (TMDB), a manually curated and web-accessible database, was developed to provide detailed, searchable descriptions of small molecular compounds found in Camellia spp. esp. in the plant Camellia sinensis and compounds in its manufactured products (different kinds of tea infusion). TMDB is currently the most complete and comprehensive curated collection of tea compounds data in the world. It contains records for more than 1393 constituents found in tea with information gathered from 364 published books, journal articles, and electronic databases. It also contains experimental 1H NMR and 13C NMR data collected from the purified reference compounds or collected from other database resources such as HMDB. TMDB interface allows users to retrieve tea compounds entries by keyword search using compound name, formula, occurrence, and CAS register number. Each entry in the TMDB contains an average of 24 separate data fields including its original plant species, compound structure, formula, molecular weight, name, CAS registry number, compound types, compound uses including healthy benefits, reference literatures, NMR, MS data, and the corresponding ID from databases such as HMDB and Pubmed. Users can also contribute novel regulatory entries by using a web-based submission page. The TMDB database is freely accessible from the URL of http://pcsb.ahau.edu.cn:8080/TCDB/index.jsp. The TMDB is designed to address the broad needs of tea biochemists, natural products chemists, nutritionists, and members of tea related research community. CONCLUSION: The TMDB database provides a solid platform for collection, standardization, and searching of compounds information found in tea. As such this database will be a comprehensive repository for tea biochemistry and tea health research community.

(Z)-5-(3,4,5-Tri-meth-oxy-styr-yl)-2,3-di-hydro-thieno[3,4-b][1,4]dioxine.

In the title compound, C17H18O5S, an analogue of the potent anti-cancer agent combretastatin A-4, the alkene C=C bond has a cis conformation and the C-C=C-C torsion angle is 9.0 (3)°. The dihedral angle between the benzene and thio-phene rings is 54.07 (4)°. The dioxene ring adopts a half-chair conformation, with the C atoms of the methyl-ene groups displaced by -0.325 (2) and 0.341 (3) Šfrom the plane of the other atoms. The C atoms of the two meta-meth-oxy groups are close to being coplanar with their attached benzene ring [displacements = -0.025 (2) and -0.196 (2) Å], whereas the C atom of the para-meth-oxy group is significantly displaced [by -1.107 (2) Å]. In the crystal, C-H⋯O hydrogen bonds link the mol-ecules into [0-11] chains, which feature two different types of R 2 (2)(6) loops.

Unraveling anti-aging mystery of green tea in C. elegans: Chemical truth and multiple mechanisms.

Tea drinking impacts aging and aging-related diseases. However, knowledge of anti-aging molecules other than the major catechins in complex tea extracts remains limited. Here we used Caenorhabditis elegans to analyze the longevity effects of tea extracts and constituents comprehensively. We found that the hot water extract of green tea prolonged lifespan and heathspan. Further, the MeOH fraction prolonged lifespan significantly longer than other fractions. Correlation analysis between mass spectroscopic data and anti-aging activity suggests that ester-type catechins (ETCs) are the major anti-aging components, including 4 common ETCs, 6 phenylpropanoid-substituted ester-type catechins (PSECs), 5 cinnamoylated catechins (CCs), 7 ester-type flavoalkaloids (ETFs), and 4 cinnamoylated flavoalkaloids (CFs). CFs (200 µM) are the strongest anti-aging ETCs (with the longest 73% lifespan extension). Green tea hot water extracts and ETCs improved healthspan by enhancing stress resistance and reducing ROS accumulation. The mechanistic study suggests that they work by multiple pathways. Moreover, ETCs modulated gut microbial homeostasis, increased the content of short-chain fatty acids, and reduced fat content. Altogether, our study provides new evidence for the anti-aging benefits of green tea and insights into a deep understanding of the chemical truth and multi-target mechanism.

Reactivity of aged biochars to the degradation of adsorbed p-nitrophenol: Role of intensity and species of persistent free radicals.

Persistent free radicals (PFRs) in biochar have been found to the transformation of organic contaminants in environment. However, there remains insufficient comprehension on the relationship of biochar aging with interfacial reactivity of PFRs to the degradation of phenolic compound in geochemical process. Herein, we studied both sorption and degradation of p-nitrophenol (PNP) on fresh and aged biochars via H2O2 aging under anoxic condition. With increasing aging extent, the enhancive proportion of O-centered radicals was observed progressively as indicated by increased g factors. The aging of PS350 annihilated the presence of PFRs in aged biochars of low-temperature, weakening PFR intensity. But, the aging of PS650 supplied more O-centered radicals for aged biochars of high-temperature, enhancing PFR intensity. This caused the decreased degradation on 5%PS350 and 15%PS350 (37.7-79.6% decline), whereas the increased degradation on 5%PS650 and 15%PS650 (33.3-55.8% increase). At similar intensity and species of PFRs, more adsorbed amount on fresh and aged biochars produced more degradation of PNP. Nevertheless, when PFR intensity of PS650 was much lower than that of PS350, despite high sorption capacity of PS650, the degradation amount of PS350 and PS650 was comparable. The results indicated that the reactivity of C-centered radicals of PS650 was stronger than that of O-centered radicals of PS350 in anoxic system. Overall, the interfacial reactivity of biochars was simultaneously regulated by the sorption capacity of biochars and intensity and species of PFRs. This work provides a deep perspective to the impact of biochar aging on the interfacial reactivity of PRFs to phenolic compound, which will be beneficial to accurately predict the fate of organic contaminant in carbon-rich environment.

Transformation of phosphorus species during phosphoric acid-assisted pyrolysis of lignocellulose.

Phosphoric acid-assisted pyrolysis (PAAP) is a pyrolysis technique with potential for the engineered and environmental application. Nevertheless, the volatilisation, immobilisation, and dissolution of phosphorus (P) species have been neglected during PAAP of lignocellulose. Therefore, we compared the transformation of P species with direct-pyrolysis and PAAP system, using multiple techniques including gas chromatography tandem mass spectrometry (GCMS) and 31P nuclear magnetic resonance (NMR). It was also investigated that the properties of pyrogenic and modified carbons obtained from lignocellulose pyrolysis at 200-650 °C. As the temperature increased, volatile P species evolved into gas-phase during PAAP, inhibiting the formation of the macromolecular volatile components. Compared with pyrogenic carbons, modified carbons with more aromatic structures experienced a higher degree of dehydration and cyclisation via catalytic crosslinking reaction. PAAP system facilitated more generation of persistent free radical (PFR) below 500 °C and the attenuation of PFR signals was observed at 500-650 °C, which may be associated with the sequestration and elimination of P species between carbon matrix. Notably, three configurations of C3PO, CPO, and COP were the major combinations of P and C elements on modified carbons. Increased gaseous P and decreased soluble P were observed with elevated temperatures in PAAP system. The species proportion of immobilised P clearly demonstrated the transformation of partial P species from inorganic to organic through pyrolysis. The immobilised P could serve as a potential sustained-release source participating in P biogeochemical cycles. These findings are fundamental for the technical design of lignocellulose pyrolysis.

Enhanced microbial degradation mediated by pyrogenic carbon toward p-nitrophenol: Role of carbon structures and iron minerals.

Pyrogenic carbon (PC) including black carbons and engineered carbons can mediate the extracellular electron transfer to facilitate the biogeochemical reaction with organic pollutants. Yet, the role of carbon structures and iron minerals on PC-mediated microbial degradation is still lacking of understanding. Herein, we studied the electrochemical properties of PCs produced from varied feedstock with regards to the mediated degradation of p-nitrophenol (PNP) by Shewanella putrefaciens CN32 in anoxic system. Mediated degradation by PCs was enhanced by facilitating extracellular electron transfer through oxygenated group and graphitic structure. Graphitic crystallites improved the electron-accepting capacity (as suggested by ID/IG and EAC) and diminished the electrochemical impedance (as suggested by Rct), contributing to PNP degradation under the anoxic system. Furthermore, more interfacial adsorption was conducive to the mediated reduction by the graphitic structure on PCs of high-temperature. In the presence of iron minerals, both hematite and goethite significantly facilitated PC-mediated degradation, which could be ascribed to the enhancement of the electron-donating capacity of microorganism and the accumulation of the reductive-state PCs by the interaction with generated Fe(II). This work paves a feasible way to the technical design on the remediation of phenolic contaminants by PC-mediated microbial degradation in environment.

Discovery of Camellia sinensis catechins as SARS-CoV-2 3CL protease inhibitors through molecular docking, intra and extra cellular assays.

BACKGROUND AND PURPOSE: Previous studies suggest that major Camellia sinensis (tea) catechins can inhibit 3-chymotrypsin-like cysteine protease (3CLpro), inspiring us to study 3CLpro inhibition of the recently discovered catechins from tea by our group. METHODS: Autodock was used to dock 3CLpro and 16 tea catechins. Further, a 3CLpro activity detection system was used to test their intra and extra cellular 3CLpro inhibitory activity. Surface plasmon resonance (SPR) was used to analyze the dissociation constant (KD) between the catechins and 3CLpro. RESULTS: Docking data suggested that 3CLpro interacted with the selected 16 catechins with low binding energy through the key amino acid residues Thr24, Thr26, Asn142, Gly143, His163, and Gln189. The selected catechins other than zijuanin D (3) and (-)-8-(5''R)-N-ethyl-2-pyrrolidinone-3-O-cinnamoylepicatechin (11) can inhibit 3CLpro intracellularly. The extracellular 3CLpro IC50 values of (-)-epicatechin 3-O-caffeoate (EC-C, 1), zijuanin C (2), etc-pyrrolidinone C and D (6), etc-pyrrolidinone A (9), (+)-gallocatechin gallate (GCG), and (-)-epicatechin gallate (ECG) are 1.58 ± 0.21, 41.2 ± 3.56, 0.90 ± 0.03, 46.71 ± 10.50, 3.38 ± 0.48, and 71.78 ± 8.36 µM, respectively. The KD values of 1, 6, and GCG are 4.29, 3.46, and 3.36 µM, respectively. CONCLUSION: Together, EC-C (1), etc-pyrrolidinone C and D (6), and GCG are strong 3CLpro inhibitors. Our results suggest that structural modification of catechins could be conducted by esterificating the 3-OH as well as changing the configuration of C-3, C-3''' or C-5''' to discover strong SARS-CoV-2 inhibitors.

Exploring new catechin derivatives as SARS-CoV-2 Mpro inhibitors from tea by molecular networking, surface plasma resonance, enzyme inhibition, induced fit docking, and metadynamics simulations.

SARS-CoV-2 Mpro (Mpro) is the critical cysteine protease in coronavirus viral replication. Tea polyphenols are effective Mpro inhibitors. Therefore, we aim to isolate and synthesize more novel tea polyphenols from Zhenghedabai (ZHDB) white tea methanol-water (MW) extracts that might inhibit COVID-19. Through molecular networking, 33 compounds were identified and divided into 5 clusters. Further, natural products molecular network (MN) analysis showed that MN1 has new phenylpropanoid-substituted ester-catechin (PSEC), and MN5 has the important basic compound type hydroxycinnamoylcatechins (HCCs). Thus, a new PSEC (1, PSEC636) was isolated, which can be further detected in 14 green tea samples. A series of HCCs were synthesized (2-6), including three new acetylated HCCs (3-5). Then we used surface plasmon resonance (SPR) to analyze the equilibrium dissociation constants (KD) for the interaction of 12 catechins and Mpro. The KD values of PSEC636 (1), EGC-C (2), and EC-CDA (3) were 2.25, 2.81, and 2.44 µM, respectively. Moreover, compounds 1, 2, and 3 showed the potential Mpro inhibition with IC50 5.95 ± 0.17, 9.09 ± 0.22, and 23.10 ± 0.69 µM, respectively. Further, we used induced fit docking (IFD), binding pose metadynamics (BPMD), and molecular dynamics (MD) to explore the stable binding pose of Mpro-1, showing that 1 could tightly bond with the amino acid residues THR26, HIS41, CYS44, TYR54, GLU166, and ASP187. The computer modeling studies reveal that the ester, acetyl, and pyrogallol groups could improve inhibitory activity. Our research suggests that these catechins are effective Mpro inhibitors, and might be developed as therapeutics against COVID-19.

N-ethyl-2-pyrrolidinone substitution enhances binding affinity between tea flavoalkaloids and human serum albumin: Greatly influenced by esterization.

Formation of catechins-human serum albumin (HSA) complex contributes to stably transporting catechins and regulating their bioavailability. Recently, a new class of catechins namely flavoalkaloids have been reported from tea. The unique structural modification with an N-ethyl-2-pyrrolidinone ring at catechins from these flavoalkaloids has raised our interest in their HSA binding affinity. Thus, we investigated the interaction between HSA and flavoalkaloids by molecular docking, UV-Vis spectroscopy (UV), fluorescence quenching approaches, and surface plasmon resonance (SPR). Thermodynamic parameters suggest that electrostatic forces contribute greatly to the interaction. The binding ability is affected by different ester group (galloyl or cinnamoyl) at 3-OH, N-ethyl-2-pyrrolidinone substituted position (C-6 or C-8), C-2, C-3 and C-5''' configurations, and hydroxyl group numbers at B ring, among which the 3-O-cinnamoyl substitution and 5'''-R configuration present the strongest contributions. UV showed slight changes in the conformation and microenvironment of HSA during the binding process. The quenching and binding constants suggest that the quenching is a static type. The small KD values (1-20 µM) detected by SPR confirmed the strong binding affinities between HSA and flavoalkaloids. Present study will help us to understand the interaction mechanism between flavoalkaloids and HSA, shedding light on structural modification of common catechins to enhance the stability, bioavailability and bioactivities.

The contrasting role of minerals in biochars in bisphenol A and sulfamethoxazole sorption.

Biochars are one of carbon-rich substances that have attracted enormous attention because of its values in energy storage, carbon sequestration, and environment remediation. Apart from the carbon structure, biochars also contain inherent mineral component and polar functional groups. However, the importance of the inherent minerals to the stability of biochars as well as the sorption of organic compounds remains unclear. In this work, the demineralized treatment by the hydrofluoric acid was employed to remove the inorganic minerals from biochars produced at 300 and 500 °C. The inorganic minerals in biochars were identified and quantified by XRD, XPS and SEM-EDS techniques. Approximately 75% of biochar minerals belonged to the Si- and Al-containing minerals, which connected with carbon skeletons. The impact of these minerals to bisphenol A (BPA) and sulfamethoxazole (SMX) sorption was investigated. The mineral removal decreased BPA sorption but increased SMX sorption. Moreover, the relative contributions of surface adsorption and partition processes were quantified for both compounds through isotherm modeling. The BPA sorption was regulated by the joint effect of adsorption and partition, while more than 82% of the SMX sorption was dominated by the partition process. Such understanding of biochar minerals and carbon structure to the migration of organic contaminants will benefit biochar production and application.

Cu2+-assisted laccase from Trametes versicolor enhanced self-polyreaction of triclosan.

Laccase-mediated humification processes (L-MHPs) can be used to polymerize and transform phenolic pollutants in water. However, the mechanism on Cu2+ impacts the self-polymerization of multi-purpose antimicrobial agent triclosan during L-MHPs is less understood. Here, the influence of divalent metal ions (DMIs) on Trametes versicolor laccase activity was investigated. Particularly, the performance of Cu2+-assisted laccase in polymerizing and transforming triclosan was systematically characterized. Compared with DMI-free, the activity of laccase was obviously accelerated with Cu2+ present due to copper is a vital component of laccase catalytic center. It was found that Cu2+-assisted laccase was effective in transforming triclosan, and the enzymatic reaction kinetic constants increased from 0.28 to 0.73 h-1 as the Cu2+ concentration increased (0-3.0 mM). Identification of intermediate products revealed that laccase oxidation predominantly generated triclosan dimers, trimers, and tetramers. The presence of Cu2+ reinforced self-polymerization of triclosan via forming more triclosan oligomers relative to the Cu2+-free, which likely attributed to the enhancement of laccase activity and stability with Cu2+ present in L-MHPs. A possible transformation mechanism was proposed as follows: Laccase initially catalyzed the oxidation of triclosan to generate phenoxy radical intermediates, which self-coupled to each other subsequently by radical-mediated CC and COC covalent binding, forming oligomers and polymers. The growth inhibitory assays of freshwater microalgae (Chlamydomonas reinhardtii and Scenedesmus obliquus) demonstrated that the self-polymerized triclosan by L-MHPs had lower toxicity than the parent compound. These findings implied that Cu2+-assisted laccase was an effective strategy for rapidly self-polyreaction and detoxication of triclosan from Cu2+-triclosan combined polluted wastewater.

The relative importance of different carbon structures in biochars to carbamazepine and bisphenol A sorption.

Biochar, a carbon-rich material, has attracted immense attention owing to its applications in soil remediation. However, the mechanisms by which heterogeneous carbon structures of biochars immobilize organic contaminants are not yet fully understood. In this study, the noncondensed aromatic components in biochars were selectively removed through bleaching. Different techniques, such as 13C nuclear magnetic resonance, were applied to characterize the biochar compositions, and thus the role of the different carbon structures in organic contaminant sorption was discussed. The aromatic carbon structures in biochars were gradually developed and evolved from noncondensed to condensed structure with increasing pyrolytic temperatures from 300 to 700 °C. Based on elemental analysis, the carbon removed by bleaching decreased from 43.9% to 5.92% with increasing temperatures. After the surface area normalization of the apparent sorption, bleaching increased the sorption of carbamazepine and bisphenol A on biochars produced at 500 °C, but not for those produced at 300 and 700 °C. Bleaching removed noncondensed aromatics and enriched condensed aromatics, which resulted in increased sorption. However, bleaching also resulted in the blockage of micropores in biochars with abundant condensed aromatics, causing decreased sorption. The apparent sorption was determined by the balancing of these two opposite effects.

Two Pairs of Isomerically New Phenylpropanoidated Epicatechin Gallates with Neuroprotective Effects on H2O2-Injured SH-SY5Y Cells from Zijuan Green Tea and Their Changes in Fresh Tea Leaves Collected from Different Months and Final Product.

Zijuan tea ( Camellia sinensis var. assamica), an anthocyanin-rich cultivar with purple leaves, is a valuable material for manufacturing tea with unique color and flavor. In this paper, four new phenylpropanoid substituted epicatechin gallates (pECGs), Zijuanins A-D (1-4), were isolated from Zijuan green tea by different column chromatography. Their structures were identified by extensive high resolution mass spectroscopy (HR-MS), nuclear magnetic resonance (NMR), and experimental and calculated circular dichroism (CD) spectroscopic analyses. Detection of the changes in fresh tea leaves collected from April to September and the final processed product by high performance liquid chromatography (HPLC)-HRMS suggested that production of compounds 1 and 2 may be enhanced by the processing procedure of Zijuan green tea. Additionally, 1-4 were proposed to be synthesized through interaction between the abundant secondary metabolite ECG and phenolic acids from tea leaves by two key steps of phenol-dienone tautomerism. 1 and 2 showed impressive activity in protecting SH-SY5Y cells against H2O2-induced damage at the concentration of 1.0 µM.

Inhibition of α-glucosidase and α-amylase by flavonoid glycosides from Lu'an GuaPian tea: molecular docking and interaction mechanism.

Green tea may favorably modulate blood glucose homeostasis, and regular consumption of green tea can prevent the development of type 2 diabetes mellitus. In this study, α-glucosidase and α-amylase inhibitory effects of the novel acylated flavonol tetraglycoside (camellikaempferoside C, 1) and 14 other flavone and flavone glycosides (FGs) isolated from Lu'an GuaPian (Camellia sinensis L.O. Kuntze) were evaluated. The kaempferol monoglycoside (15) showed inhibitory activity against α-glucosidase with IC50 at 40.02 ± 4.61 µM, and kaempferol diglycoside (13) showed α-amylase inhibition with IC50 at 0.09 ± 0.02 µM. Further, inhibitory mechanisms of FGs 15 and 13 were studied by molecular docking analysis and fluorescence spectrometry. Molecular docking suggested that FG 15 interacted with α-glucosidase mainly by hydrogen bonding, which was the same interaction force between FG 13 and α-amylase. Intrinsic fluorescence of α-glucosidase and α-amylase was quenched by 15 and 13, respectively, through a static quenching mechanism. The spontaneous formation of 15-α-glucosidase and 13-α-amylase complexes was driven by van der Waals forces and hydrogen bonding. The present study provides new insight into the potential application of Lu'an GuaPian green tea as a functional food ingredient to regulate postprandial hyperglycemia through inhibition of α-glucosidase/α-amylase by FGs, particularly the mono- and di- glycosides of kaempferol.

Interaction between Ester-Type Tea Catechins and Neutrophil Gelatinase-Associated Lipocalin: Inhibitory Mechanism.

Tea is thought to alleviate neurotoxicity due to the antioxidative effect of ester-type tea catechins (ETC). Neutrophil gelatinase-associated lipocalin (NGAL) can sensitize ß-amyloid (Aß) induced neurotoxicity, and inhibitors of NGAL may relieve associated symptoms. As such, the interactions of ETC with NGAL were investigated by fluorescence spectrometry and molecular simulation. NGAL fluorescence is quenched regularly when being added with six processing types of tea infusion (SPTT) and ETC. Thermodynamic analyses suggest that ETC with more catechol moieties has a stronger binding capacity with NGAL especially in the presence of Fe3+. (-)-Epicatechin 3-O-caffeoate (ECC), a natural product isolated from Zijuan green tea, shows the strongest binding ability with NGAL (Kd = 15.21 ± 8.68 nM in the presence of Fe3+). All ETC are effective in protecting nerve cells against H2O2 or Aß1-42 induced injury. The inhibitory mechanism of ETC against NGAL supports its potential use in attenuation of neurotoxicity.