Epicatechin attenuates lead(Pb)-induced cognitive impairment in mice:regulation on Nrf2 signaling pathway,and interference on the interaction between Pb with albumin

Epicatechin(EC)was used in this study to antagonize the cognitive dysfunction caused by lead(Pb)exposure in mice.Eight-week-old male Kunming mice were treated with PbCl_(2)(20 mg/kg)and/or EC(50 mg/kg)by gavage administration for 4 weeks.Morris water maze test showed that EC could improve memory dysfunction induced by Pb.EC antagonized Ca^(2+)overload,activated Nrf2 signaling pathway and reduced the accumulation of Pb in the brain and serum,which suggested that EC might alter Pb distribution in mice.In vitro,spectroscopic analysis,potentiometric titration and docking studies were applied to inquiry into the interaction between bovine serum albumin(BSA)and Pb^(2+)in presence or absence of EC.EC was proved to chelate Pb^(2+)and reduced the interaction between BSA and Pb^(2+).In summary,EC might protect Pb-induced cognitive impairment by activating Nrf2 signaling pathway,and suppressing Pb accumulation via interference on the binding of Pb to albumin.

Optimization of Extraction Process of Fagopyri Dibotryis Rhizoma by Orthogonal Experiment

[Objectives]To optimize the water extraction process of Fagopyri Dibotryis Rhizoma.[Methods]The entropy weight method was used to determine the weight of epicatechin extraction rate and dry extract rate and calculate the comprehensive score.The water extraction process of Fagopyri Dibotryis Rhizoma was optimized by orthogonal design with the comprehensive score as the indicator and the amount of water,extraction time and extraction times as the factors.[Results]The optimum extraction process of Fagopyri Dibotryis Rhizoma was as follows:adding 10 times of water,extracting 3 times,and extracting for 60 min each time.[Conclusions]The optimized extraction process is stable and feasible,and can be used for the extraction of Fagopyri Dibotryis Rhizoma.

Adsorption Isotherms of Quercetin and Catechin Compounds on Quercetin-MIP

A molecular imprinted polymer（MIP） was prepared with quereetin as the template and methaerylie acid（MAA） as the functional monomer. Aeetonitrile and methanol were used as the porogen with ethylene glycol dimethaerylate （EGDMA） as the erosslinker and 2,2＇-azobis （ isobutyronitrile ） （ AIBN ） as the initiator. The experimental parameters of the equilibrium isotherms were estimated via linear and nonlinear regression analyses. The linear equadon as the functions of the adsorption concentration of the single compound in its solution and the competitive adsorption of the single compound in its mixed compounds solution was then expressed, and the adsorption equilibrium data were correlated to Langmuir and Freundlich isotherm models. The mixture compounds show competitive adsorption on the specific binding sites of quereetin-MIP. Furthermore, the competitive Langmnir isotherms were applied to the mixture compounds. The adsorption concentrations of quercetin, （＋）eatechin（＋C）, and （-）epieateehin（EC） on the quercetin molecular imprinted polymer were compared. The quercetin-imprinted polymer shows extraordinarily higher adsorption ability for quercetin than for the two eateehin compounds that were also assessed.

Differential Anticancer Effect of an Apple Extract (Applephenon^&reg), Polyphenols and Isoflavones on Normal Human Keratinocytes and Epidermoid Cancer Cells

Applephenon&reg, a purified extract prepared from green apples, was examined for its cytotoxicity and inhibitory effects on the proliferation of cultures of normal human keratinocytes and several epidermoid cancer cell lines. Our HPLC studies demonstrated a high content of phenolic compounds (>65%), including catechin, epicatechin, caffeic acid and phloretin as well as polyphenols such as proanthocyanidins. Applephenon&reg demonstrated a greater cytotoxic effect against HeLa, A431 cancer cell lines and HaCaT, an immortalized keratinocyte cell line than serum-free cultures of proliferating normal human keratinocytes (NHK). Proliferation of NHK was inhibited at concentrations above 0.0013% while concentrations above 0.005% were cytotoxic. By contrast, Applephenon&reg solutions above 0.00025% killed each of the cancer cell lines. Treated cells displayed increased intercellular separation and evidence of keratinizing stratification. We also tested the effect of epicatechin, and two isoflavonoids, genistein and daidzein, on cancer cell lines. Hela cells were more sensitive to epicatechin and genistein inhibition of cell growth and cytotoxicity than were NHK. Daidzein at these concentrations had little effect on cancer cells. These results indicate that Applephenon&reg and some of its phenolic components have selective anticancer activity.

Catechin and Epicatechin. What’s the More Reactive?

Catechin and epicatechin are two isomeric flavonoids. Despite the vital properties highlighted by numerous scientific studies, very little data is available on the intrinsic reactivity of these compounds. To provide more details on the stability and reactivity of catechin and epicatechin, this study is performed by means of theoretical calculation methods. For this purpose, geometry optimizations and frequency calculations at the B3LYP/6-31 + G (d, p) level of theory has been carried out and Natural Bond Orbital (NBO) analysis and VEDA (Vibrational Energy Distribution Analysis). The geometric and energy parameters and NBO analysis show that catechin appears more stable than epicatechin. The hydroxyl group position on the ring C of the catechol structure represents a factor that influences this relative stability. The global and local reactivity parameters reveal that epicatechin becomes more reactive than catechin. They indicate that their hydroxyl groups correspond to their most receptive sites. Fukui indices, VEDA and acidity study establish that O28–H29 remains the most reactive.

Efficient semisynthetic approach for large preparation of procyanidin C1 through degradation of grape seed polymeric procyanidins

An efficient method for producing trimeric procyanidin C1(PCC1)was developed through degradation of grape seed polymeric procyanidins(PPCs),using epicatechin(EC)as nucleophile and hydrochloric acid as catalyst.With the yield of PCC1 as the evaluation index,the degradation conditions were optimized by Box-Behnken Design(BBD)based on the results of single-factor experiments.The results showed that the optimal conditions were reaction temperature of 40℃,the ratio of EC and PPCs 2:1,acidity of 0.10 mol/L,and reaction time of 20 min.The yield of PCC1 reached up to 17.7 mg by only one-step degradation of 3 g PPCs.This work proposed a new method for large preparation of PCC1 from waste grape seed polymeric procyanidins.

RNA-seq analysis of protective effect of epicatechin gallate on H_(2)O_(2)-induced oxidative injury in PC12 cells

Epicatechin gallate(ECG)is one of the polyphenolic compounds and has attracted much attention due to its various bioactivities.In this study,the neuroprotective eff ect of ECG against H_(2)O_(2)-induced oxidative injury in PC12 cells as well as the possible mechanisms were investigated.Cell viability was determined by MTT assay.The differentially expressed genes(DEGs),GO enrichment,and KEGG enrichment were analyzed to explore the mechanism of ECG against H_(2)O_(2)-induced oxidative injury by using the RNA-seq method.Finally,the change in the cell cycle was analyzed by fl ow cytometry.H_(2)O_(2)(400-1200μmol/L)inhibited the cell viability in a concentration-dependent manner.ECG(6-150μmol/L)eff ectively attenuated the H_(2)O_(2)-induced decrease in cell viability.RNA-seq analysis showed that ECG regulated 1058 coexpressed DEGs.GO enrichment analysis showed that the cellular component was the dominant group after ECG treatment.KEGG analysis showed that the cell cycle,fanconi anemia pathway,and homologous recombination were the important pathways for ECG in improving H_(2)O_(2)-induced oxidative injury and 28 coexpressed DEGs in the cell cycle pathway were summarized.Finally,cell cycle analysis also proved that ECG improved H_(2)O_(2)-induced cell cycle arrest in the G2/M phase.Our present study demonstrated that ECG attenuated H_(2)O_(2)-induced neurologic oxidative damage by multiple modulatory mechanisms at the molecular transcription level.These fi ndings provide new insights for further study of the molecular mechanism of the neuroprotection of ECG.

Excretion of Four Catechins in Tea Polyphenols in Rats

Objective To investigate excretion profiles of the four major anti-oxidant active catechins, (-) epigallo-catechin-3-gallate (EGCG), (-) epicatechin-3-gallate (ECG), (-) epigallocatechin (EGC), and epicatechin (EC) in tea polyphenols (TP) in rats in order to provide experimental data for clinical uses and development of TP as a novel drug. Methods The above four catechins in urine, bile, and feces were simultaneously determined by high performance liquid chromatography coupled with ultraviolet absorption detector (HPLC-UV) assay with a binary gradient elution. The samples were extracted by ethyl acetate prior to HPLC. The quantification was carried out by peak area internal standard method. Following iv dosing TP 100 mg/kg to rats, the samples were collected at different time intervals up to 8 h (urine and bile) and 24 h (feces). Results The urinary Ae, 0-8 h (cumulative excretion amount over 8 h) of EGCG, ECG, EGC, and EC were, on the average, 150.83, 30.75, 116.69, and 254.56 μg, corresponding to fe, 0-8 h (cumulative excretion fraction of dose over 8 h) of 1.45%, 0.84%, 7.88%, and 10.73%, respectively; the biliary Ae, 0-8 h were 12.61, 42.64, 6.61, and 1.24 μg, corresponding to the fe, 0-8 h of 0.12%, 1.16%, 0.45%, and 0.053%,respectively. For fecal excretion, only EGCG and EGC were detected with Ae, 0-24 h of 7.38 μg (fe, 0-24 h of 0.07%) and 157 μg (fe, 0-24 h of 9.99 %), respectively. The fe, total (the total fe of 3 excretory routes) were 18.32%, 10.78%, 2.00%, and 1.64% for EGC, EC, ECG, and EGCG, respectively. Conclusion EGCG and EC are mainly excreted in urine, ECG in bile, and EGC in feces by reference to their Ae and fe. The excretion of the four catechins based on fe, total is ranked in order of EGC > EC > ECG > EGCG. Only small amount of four catechins are recovered in urine, bile, and feces, indicating an extensive metabolic conversion of catechins in the rat body.

Interaction of epicatechin with bovine serum albumin using fluorescence quenching combined with chemometrics

The interaction between BSA and epicatechin was studied using fluorescence quenching titrations combined with trilinear decomposition method and excitation-emission matrix(EEM)fluorescence.The resolved spectra were highly similar with the actual ones which indicated that the resolved results were reliable.The relevant parameters of the binding process were obtained by quantifying each substance in the complicated mixtures in situ.The quenching was static quenching,epicatechin had a weak interaction with BSA and the binding site was one.The total concentration and the free concentration of quenchers had different effect on the system.The results demonstrated that the method exploited in this article is a useful tool to investigate complicated interactions,avoiding complicated pretreatment and simplify experimental procedure.

In silico study unfolds inhibitory potential of epicatechin gallate against SARS-CoV-2 entry and replication within the host cell

Coronavirus disease-19(COVID-19)is the ongoing pandemic affecting millions of people worldwide.Several vaccine candidates have been designed and developed for the causative virus,SARS-CoV-2.However high mu-tation rate in the viral genome and the emergence of new variants have challenged the effectiveness of these vaccines developed for previous strains.Hence,screening and identification of anti-SARS-CoV-2 agents having multi-target potency would be more impactful in the prevention of the disease.Epicatechin gallate(ECG)is a green tea polyphenol having various medicinal properties,including anti-oxidative and anti-inflammatory effects.However its role as anti-SARS-CoV-2 agent is not clear.Hence the present in silico study aims to investigate the binding potential of ECG with several proteins which are critical to SARS-CoV-2 entry and replication within the host cell.Molecular docking analyses have revealed that ECG could potentially block several amino acid residues of entry factors in host cells,spike protein,and many non-structural proteins through Hydrogen bonds and hy-drophobic interactions.Such interactions with vital proteins could inhibit SARS-CoV-2 entry and its subsequent replication into the host.Therefore,ECG could be a potential therapeutic agent for the prevention of COVID-19.However,the findings of the present study demand further validation in animal models.