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Berberine (BBR) is the main pharmacological active ingredient of Coptidis, which has hypoglycemic effect, but its clinical application is limited due to its poor oral bioavailability. Polyphenols, derived from cinnamon, are beneficial for type 2 diabetes mellitus (T2DM). The combination of both may have an additive effect. The aim of this study was to investigate the hypoglycemic effect and mechanism of combined medication in diabetic rats. The modeling rats were randomly divided into 5 groups (berberine group, cinnamon group, combined group, metformin group, diabetic control group) and normal control group. The animal experiments were approved by the Animal Ethics Committee (approval number: HMUIRB2022003). The subjects were given orally, and the control group was given equal volume solvent and body weight was measured weekly. Thirty days after administration, oral glucose tolerance test and insulin sensitivity test were performed, and fasting blood glucose (FBG), glycated serum protein (GSP), and serum insulin (INS) levels were detected; high-throughput sequencing technology was used to detect intestinal microbiota structure; real-time quantitative PCR (RT-qPCR) and Western blot were used to detect G protein-coupled receptor 5 (TGR5) and glucagon-like peptide-1 (GLP-1) expression levels. The results showed that, compared with the diabetic control group, the levels of FBG (P < 0.01) and GSP (P < 0.01) in the combined group were lower, and the insulin resistance was improved, which was better than that in the berberine group. Combined treatment increased the relative abundance of Bacteroides, Prevotella and Lactobacillus, reversed the decrease in Lactobacillus in the berberine alone induction group, and the combination of the two could promote the expression of TGR5 and GLP-1. In summary, the combined application of cinnamon and berberine can regulate glucose metabolism better than the application of berberine alone. Berberine combined with cinnamon can improve the function of pancreatic islet β cells in diabetes mellitus type 2 rats by changing the intestinal microbiota, increasing the expression of TGR5 and GLP-1 proteins, and thereby better regulating glucose metabolism.
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The goal of this work was to investigate the antidepressant fraction from Radix Paeoniae Alba and identify its major chemical constituents. Corticosterone injured rat phaeochromocytoma (PC12) cells and behavioral despair depression models of mice were used to evaluate the antidepressant effects of Radix Paeoniae Alba (Bai-Shao) ethanol extract (BS-E) and its three fractions (BS-10E, BS-60E, BS-95E) isolated by macroporous resin column chromatography. Animal experimental procedures were approved by the Animal Ethics Committee of the Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College (approval No.: SLXD-20210618051). The results showed that BS-E, BS-10E and BS-60E had protective effects against PC12 cells injury induced by corticosterone, among which BS-60E had the strongest protective effect. BS-60E could significantly shorten the time of forced swimming and tail suspension in despair depression models of mice, and was identified as the antidepressant fraction of Radix Paeoniae Alba. The major chemical constituents in the antidepressant fraction were identified by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), and their proposed fragmentation pathways in MS spectra were deduced. A total of 79 chemical constituents were identified from BS-60E, including 36 monoterpenes, 34 polyphenols, 6 oligosaccharides, and 3 other constituents, and monoterpenes and polyphenols may be major effective constituents of BS-60E.
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Microglial surveillance plays an essential role in clearing misfolded proteins such as amyloid-beta, tau, and α-synuclein aggregates in neurodegenerative diseases. However, due to the complex structure and ambiguous pathogenic species of the misfolded proteins, a universal approach to remove the misfolded proteins remains unavailable. Here, we found that a polyphenol, α-mangostin, reprogrammed metabolism in the disease-associated microglia through shifting glycolysis to oxidative phosphorylation, which holistically rejuvenated microglial surveillance capacity to enhance microglial phagocytosis and autophagy-mediated degradation of multiple misfolded proteins. Nanoformulation of α-mangostin efficiently delivered α-mangostin to microglia, relieved the reactive status and rejuvenated the misfolded-proteins clearance capacity of microglia, which thus impressively relieved the neuropathological changes in both Alzheimer's disease and Parkinson's disease model mice. These findings provide direct evidences for the concept of rejuvenating microglial surveillance of multiple misfolded proteins through metabolic reprogramming, and demonstrate nanoformulated α-mangostin as a potential and universal therapy against neurodegenerative diseases.
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AIM:To investigate the effects of Prunus cerasifera Ehrh.polyphenol extract(PCE)on glucose metabolism in insulin-resistant HepG2(IR-HepG2)cells.METHODS:An IR-HepG2 cells model was established using high sugar and high insulin induction;a control group,an insulin resistance(IR)model group,a PCE treatment group with different concentrations,and a metformin treatment group were set up.Cells were treated with 50,100,200,and 400 mg/L of polyphenol extract for 24 hours,respectively,to detect their effects on cellular glucose consumption.At the same time,use thiazole blue(MTT)colorimetry to evaluate cell viability.Real-time PCR and Western blot were used to detect expression of glucose metabolism-related mRNA and proteins.RESULTS:Prunus cerasifera Ehrh.polyphenol ex-tract can significantly increase glucose consumption in IR-HepG2 cells,and cell activity tends to increase and then decline as its concentration increases;At mRNA and protein levels,in addition to significantly increasing the expression of phos-phorylated adenosine monophosphate activated protein kinase(p-AMPK)protein in IR-HepG2 cells,it also decrease gly-cogen synthase kinase 3β(GSK3β)in glycogen synthesis and gluconeogenesis pathways,forked protein O1(FoxO1),peroxisome proliferator activates gamma co-receptor activation 1-alpha(PGC-1α),glucose-6-phosphatase(G6Pase)and phosphoenol pyruvate carboxykinase(PEPCK)expression.CONCLUSION:Prunus cerasifera Ehrh.polyphenol extract can activate the AMPK signaling pathway in IR-HepG2 cells,to further inhibit gluconeogenesis and glycogen synthesis pathways,thereby improving the insulin resistance state of HepG2 cells.
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Phyllanthi Fructus is a highly unique medicine and food homologous item, which exhibits distinctive flavor, notable nutritional value, and abundant pharmacological activity. It has enormous potential in the creation of health products and pharmaceuticals. However, due to the unique laws of quality formation and transfer of Phyllanthi Fructus, its appearance, shape, chemical compositions, nutrients, and sensory flavors are frequently greatly influenced by botanical resources, the processing and storage conditions. As a result, the current quality evaluation model is difficult to meet the needs of Phyllanthi Fructus as a medicine and food homologous item in the development of diversified products. This paper constructs the hierarchical utilization mode of Phyllanthi Fructus based on its unique quality formation and transmission laws, explores the quality evaluation model for food-oriented use and medicinal-oriented use, respectively, and systematically describes the quality evaluation idea under diversified application scenarios. This paper aims to serve as a reference for the construction of a quality evaluation model suitable for the medicine and food homologous item of Phyllanthi Fructus.
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Abstract This study investigated the changes in the ingredients in Fallopia multiflora Thunb. Haraldson (FMT) root after processing it with different methods such as soaking, stewing, and steaming or combined methods. The total polyphenol, 2,3,5,4'-tetrahydroxystilben-2-O-ß-D-glucoside (THSG), and physcion contents in FMT products after processing were determined using high-performance liquid chromatography (HPLC) and ultraviolet-visible (UV-VIS) methods. The results demonstrated that the processing method and time significantly affected the contents of polyphenol, THSG, and physcion. The physcion and total polyphenol content increased or decreased during processing depending upon the processing time, while the THSG content gradually decreased with an increase in the processing time. The content of physcion (a substance that can cause liver toxicity) was analysed, and the suitable conditions for processing of the FMT products were determined as initial soaking in rice swill for 24 h and subsequent stewing with black beans and water for 12 h
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Fallopia multiflora/genética , Métodos , Cromatografia Líquida de Alta Pressão/métodos , Polifenóis/agonistas , Fígado/anormalidadesRESUMO
OBJECTIVE To study the vasorelaxant effects and mechanism of polyphenol compound 2,4′,5′-trihydroxy-5,2′- dibromo-diphenyl-methanone(LM49)on isolated aortic rings of rats. METHODS Thoracic aortic vascular rings of rats were collected. Using the diastolic rate as index , the effects of different concentrations of LM 49 on endothelium-intact and endothelium-denuded aortic rings pre-contracted by norepinephrine (NE,1×10-6 mol/L)or KCl (60 mmol/L)were investigated. After pre-culturing vascular rings by nitric oxide synthase inhibitor L-nitro-arginine methyl ester (L-NAME,0.1 mmol/L) and cyclooxygenase inhibitor indomethacin (1×10-5 mol/L),as well as pre-culturing vascular rings by 4 potassium channel blockers [BaCl 2(0.1 mmol/L),tetraethylammonium(TEA,5 mmol/L),4-aminopyridine(4-AP,0.1 mmol/L)and glibenclamide (1×10-5 mol/L)],the vasorelaxant effect of different concentrations of LM 49 on the vascular rings were investigated by using the same method. With the percentage of vasoconstriction as the index ,using KCl (60 mmol/L),NE(1×10-6 mol/L),calcium channel blocker verapamil (1×10-6 mol/L)and sarcoplasmic Δ 基金项目 重大新药创制国家科技重大专项 (No.2018ZX097- reticulum Ca 2 +-adenosine triphosphate (ATP) enzyme pump inhibitor thacarotene (TG,1×10-6 mol/L)to induce the release of calcium in vascular rings in the absence of calcium. CaCl was added cumulatively ,and the effect of LM 49 on the cxyw06,vasoconstriction caused by calcium influx induced by CaCl 2 was investigated. RESULTS 3×10-6,5×10-6,1×10-5 mol/L LM49 had a significant relaxation effect on NE and KCl precontracted vascular rings (P<0.01); whether the endothelium was removed or not had no significant effect on the vasodilation of LM 49(P>0.05). After L-NAME ,indomethacin, TEA and 4-AP was pre-incubated ,different concentrations of LM 49 had no significant effects on aortic rings precontracted by NE (P>0.05). Glibenclamide and BaCl 2 could inhibit the vasorelaxant effects of LM 49 on aortic rings precontracted by NE (P<0.01). In the absence of calcium ,LM49 could inhibit the contraction caused by calcium influx induced by accumulated CaCl 2 after pre-incubation with KCl ,NE,verapamil and TG (P<0.01). CONCLUSIONS LM49 evokes significant relaxation of isolated aortic vascular rings without endothelium dependence ;the mechanism of which is inducing ATP-sensitive potassium channel , inward rectifier potassium channel open and restraining extracellular Ca 2 + influx via voltage-gated calcium channel , receptor-operated calcium channel and store-operated calcium channel.
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Acute lung injury(ALI)is a critical respiratory disorder characterized by progressive respiratory failure with high morbidity and mortality.In addition to respiratory support, ALI still lacks effective medications.Natural polyphenol, a class of natural compounds widely found in human daily food, have been proven to possess anti-inflammatory, anti-oxidant, regulating cell death, anti-pathogen infection, and exert therapeutic effects on ALI.This article reviews the pathogenesis of ALI and the mechanism of natural polyphenols in the treatment of ALI, aiming to provide a new direction for the treatment of ALI.
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Abstract Obesity and dyslipidemia are conditions often associated with cardiovascular risk, inflammation, oxidative stress, and death. Thus, a new approach has been highlighted to promote research and development of pharmacological tools derived from natural sources. Among the most widely studied groups of substances, polyphenols such as tyramine stand out. This study investigated hypolipidemic and anti-obesity properties of tyramine. Oral toxicity evaluation, models of dyslipidemia and obesity were used. To induce dyslipidemia, Poloxamer-407 (P-407) was administered intraperitoneally. In the hypercholesterolemic and obesity model, specific diet and oral tyramine were provided. After 24h of P-407 administration, tyramine 2 mg/kg (T2) decreased triglycerides (TG) (2057.0 ± 158.5 mg/dL vs. 2838 ± 168.3 mg/dL). After 48h, TG were decreased by T2 (453.0 ± 35.47 vs. 760.2 ± 41.86 mg/dL) and 4 mg/kg (T4) (605.8 ± 26.61 760.2 ± 41.86 mg/dL). T2 reduced total cholesterol (TC) after 24h (309.0 ± 11.17 mg/dL vs. 399.7 ± 15.7 mg/dL); After 48h, 1 mg/kg (T1) (220.5 ± 12.78 mg/dL), T2 (205.8 ± 7.1 mg/dL) and T4 (216.8 ± 12.79 mg/dL), compared to P-407 (275.5 ± 12.1 mg/dL). The treatment decreased thiobarbituric acid reactive substances and nitrite in liver, increased superoxide dismutase, reduced the diet-induced dyslipidemia, decreasing TC around 15%. Tyramine reduced body mass, glucose, and TC after hypercaloric feed. Treatment with 5 mg/L (0.46 ± 0.04 ng/dL) and 10 mg/L (0.44 ± 0.02 ng/dL) reduced plasma insulin (1.18 ± 0.23 ng/dL). Tyramine increased adiponectin at 5 mg/L (1.02 ± 0.02 vs. 0.83 ± 0.02 ng/mL) and 10mg/L (0.96 ± 0.04 ng/mL). In conclusion, tyramine has low toxicity in rodents, has antioxidant effect, reduces plasma triglycerides and cholesterol levels. However, further studies should be conducted in rodents and non-rodents to better understand the pharmacodynamic and pharmacokinetic properties of tyramine
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Tiramina/efeitos adversos , Hipolipemiantes/farmacologia , Obesidade/classificação , Colesterol/farmacologia , Hiperlipidemias/complicaçõesRESUMO
BACKGROUND: In fish farming, the plant extracts containing antioxidant compounds have been added to the diet for enhancing pathogen resistance. In vitro studies evaluating the antioxidant effect of herbal extracts on fish cell models have focused on ROS production and the respiratory burst mechanism. However, the effects on enzymatic antioxidant defense on salmon leukocytes have not been evaluated. This study aims to evaluate the enzymatic antioxidant defense and ROS-induced cell damage in Salmon Head Kidney-1 (SHK-1) cell line exposed to polyphenol-enriched extract from Sambucus nigra flowers. RESULTS: Firstly, the Total Reactive Antioxidant Power (TRAP) assay of elderflower polyphenol (EP) was evaluated, showing 459 and 489 times more active than gallic acid and butyl hydroxy toluene (BHT), respectively. The toxic effect of EP on salmon cells was not significant at concentrations below 120 mg/ mL and no hemolysis activity was observed between 20 and 400 mg/mL. The treatment of SHK-1 cell line with EP decreased both the lipid peroxidation and protein oxidation induced by H2O2, which could be associated with decreasing oxidative stress in the SHK-1 cells since the GSH/GSSG ratio increased when only EP was added. CONCLUSIONS: These results suggest that plant extracts enriched with polyphenols could improve the enzymatic antioxidant defense of salmon leukocytes and protect the cells against ROS-induced cell damage
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Salmão , Extratos Vegetais/farmacologia , Sambucus nigra/química , Polifenóis/farmacologia , Peroxidação de Lipídeos , Sequestradores de Radicais Livres , Espécies Reativas de Oxigênio , Aquicultura , Estresse Oxidativo , Salmo salar , Resistência à Doença , Leucócitos , AntioxidantesRESUMO
Salidroside (SAL) is a phenolic substance with high solubility and low permeability, which make it easy to cause the efflux effect of P-glycoprotein and degradation of intestinal flora, resulting in lower bioavailability. The aim of this study was to develop and optimize a water-in-oil nanoemulsion of SAL (w/o SAL-N) to explore its suitability in oral drug delivery systems. In this work, SAL-N was successfully prepared by water titration method at K
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Developing tumor-specific drug delivery systems with minimized off-target cargo leakage remains an enduring challenge. In this study, inspired from the natural cryptobiosis explored by certain organisms and stimuli-responsive polyphenol‒metal coordination chemistry, doxorubicin (DOX)-conjugated gelatin nanoparticles with protective shells formed by complex of tannic acid and Fe
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To investigate the enzyme properties of the black sesame polyphenol oxidase (BsPPO), a synthesized Bsppo gene was cloned into the vector pMAL-c5x and expressed in E. coli. Subsequently, the MBP fusion label in the recombinant protein was removed by protease digestion after affinity purification. The synthesized Bsppo gene contained 1 752 bp which encodes 585 amino acids with a deduced molecular weight of 65.3 kDa. Transformation of the recombinant vector into E. coli BL21(DE3) resulted in soluble expression of the fusion protein MBP-BsPPO. The enzymatic properties of the recombinant BsPPO was investigated after MBP fusion tag excision followed by affinity purification. The results demonstrated that the optimal temperature and pH for BsPPO was 25°C and 4.0, respectively. BsPPO exhibited a good stability under low temperature and acidic environment. Low-intensity short-term light exposure increased the activity of BsPPO. Cu²⁺ could improve the activity of BsPPO while Zn²⁺ and Ca²⁺ showed the opposite effect. BsPPO could catalyze the oxidation of monophenols, diphenols, and triphenols, and exhibited good catalytic activity on l-tyrosine and vanillic acid. Moreover, BsPPO exhibited high catalytic activity on black sesame metabolites, including 2-methoxy cinnamic acid, indole-3-carboxylic acid and phloretin. These results may serve as a basis for further characterization of BsPPO.
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Catecol Oxidase/genética , Clonagem Molecular , Escherichia coli/metabolismo , Proteínas Recombinantes/genética , Sesamum/genéticaRESUMO
There is no consensus on the content, accumulation, transformation and content determination methods of phenolic acids in fresh Salvia miltiorrhiza. In order to find out the true content of phenolic acids in fresh S. miltiorrhiza, a variety of treatment me-thods were used in this study to prepare sample solution. The content changes of phenolic acids in S. miltiorrhiza samples with different dehydration rates were investigated during drying and shade drying processes. Polyphenol oxidase(PPO) of S. miltiorrhiza was extracted and purified by ammonium sulfate precipitation and dialysis to investigate the enzymatic properties. The content of rosmarinic acid, lithosperic acid and S. nolic acid B in S. miltiorrhiza was determined by UPLC. The results showed that the content of phenolic acids in fresh S. miltiorrhiza was highest when it was homogenized with 1 mol·L~(-1) HCl solution or 1 mol·L~(-1) HCl methanol solution. There was no significant difference in the content of phenolic acids in S. miltiorrhiza with different dehydration rates, indicating that there was no correlation between phenolic acid content and dehydration rate. The optimum pH of S. miltiorrhiza PPO was 7.6 and the optimum temperature was 40 ℃. With catechol as substrate, S. miltiorrhiza PPO had the enzymatic browning reaction which was in compliance with Michaelis equation, with Michaelis constant K_m of 0.12 mol·L~(-1) and V_(max) of 588.23 U·min~(-1). The inhibitory effect of citric acid, disodium ethylenediamine tetraacetate, ascorbic acid and sodium sulfite on S. miltiorrhiza PPO increased with the increase of inhibitor concentration, and sodium sulfite showed the strongest inhibitory effect. The present study proved that there were a large number of phenolic acids in fresh S. miltiorrhiza, which were the secondary metabolite of primitive accumulation during the growth of S. miltiorrhiza, rather than the induced product of postharvest drying and dehydration stress. This study has reference value and significance for the cultivation, harvest and processing of S. miltiorrhiza.
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Catecol Oxidase , Dessecação , Hidroxibenzoatos , Raízes de Plantas , Salvia miltiorrhizaRESUMO
The activity and kinetic parameters of the enzyme polyphenol oxidase (PPO) in crude extracts of the fruits ofParadise apple (Pyrus malus L.), cucumber (Cucumis sativus L.), squash (Cucurbita pepo L.), pomegranaterind and seeds (Punica granatum L.), and the leaf of cactus (Opuntia ficus-indica (L.) were investigated. Thehighest browning intensity was found in P. apple, followed by pomegranate seeds and cucumber fruits. Theoptimum activity of the enzyme was shown at pH 7.0 for P. apple and cucurbita, but it was found at pH 6.0 forthe rind and seeds of the pomegranate. The PPO of cactus leaf and cucumber fruits showed two peaks at pH 4.0and 7.0. On the other hand, the optimal temperature found for PPO activity in these plants was around 40°C.A strong correlation with value (R) = 0.9485 between the browning intensity and PPO activity in all studiedsamples was obtained.
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Abstract Ceratobasidium ramicola is a fitopathogenic fungus that harmful and causes various levels of damage on several types of forestry and horticultural crops. The purpose of this study was to examine the effect of Acacia villosa, Myristica fragrans, Acacia mangium and Calliandra calothyrsus leaf extracts as tannin sources related to the in vitro inhibition of Ceratobasidium ramicola growth. The in vitro inhibition was performed by employing solid potato dextrose agar (PDA) medium to obtain the radial inhibition, while liquid potato dextrose broth (PDB) medium was used to obtain the biomass inhibition. Experimental design was based on in time nested-completely randomized design and statistical analysis was carried out with SAS software version 9.1. The result of radial growth inhibition of Ceratobasidium ramicola showed that tannin extracts of A. mangium and M. Fragrans were not significantly different to each other. Treatment of tannin extracts from A. villosa, M. fragrans, A. mangium and C. Calothyrsus with a concentration of 1% were significantly different with other concentrations and resulted the greatest inhibition values. Tannin extract of A. Mangium at 1% concentration produced the greatest radial inhibition by 33.2%. In most cases, the effective inhibition from tannin extract occurred at the 24h of incubation. The greatest biomass inhibition was produced on 1% tannin extract of A. mangium by 64.3%, while the lowest was produced from 1% tannin extract of M. fragrans by 27.0%.
Resumo Ceratobasidium ramicola é um fungo fitopatogênico prejudicial que causa vários danos em diversas culturas florestais e agrícolas. O objetivo deste estudo foi examinar o efeito dos extratos de folhas de Acacia villosa, Myristica fragrans, Acacia mangium e Calliandra calothyrsus como fontes de tanino relacionadas à inibição do crescimento in vitro de Ceratobasidium ramicola. A inibição in vitro foi realizada empregando o meio de ágar batata dextrose para obter a inibição radial, enquanto o meio de caldo de batata dextrose líquido foi usado para obter a inibição da biomassa. O projeto experimental foi fundamentado in time. O delineamento utilizado foi inteiramente casualizado, e a análise estatística foi realizada com o software SAS, versão 9.1. O resultado da inibição do crescimento radial de Ceratobasidium ramicola mostrou que os extratos de tanino de A. mangium e M. fragrans não diferiram significativamente entre si. Os tratamentos de extratos de tanino de A. villosa, M. fragrans, A. mangium e C. Calothyrsus, utilizando uma concentração de 1%, foram significativamente diferentes em comparação com outras concentrações e resultaram em maiores valores de inibição. O extrato de tanino de A. mangium a 1% de concentração produziu a maior inibição radial, com taxa de 33,2%. Na maioria dos casos, a inibição efetiva do extrato de tanino ocorreu em 24 horas de incubação. A maior inibição de biomassa foi produzida em 1% de extrato de tanino de A. Mangium, com taxa de 64,3%, enquanto a menor foi produzida a partir de 1% de extrato de tanino de M. fragrans, com taxa de 27%.
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Myristica , Acacia , Fabaceae , Taninos , Extratos VegetaisRESUMO
Tea is one of the most widely consumed beverages in the world and is recognized as having numerous beneficialhealth effects due to its phenolic content. The current study aimed to evaluate and compare the phenolic andflavonoid content of two forms of Twining English Breakfast Tea: tea bags (TBs) and opened tea bags (OTBs).The antioxidant activity was also assessed using DPPH, FRAP, and CUPRAC assays. The results obtainedrevealed that the OTB infusion exhibited significantly higher phenolic and flavonoid content than the TB infusionat 5 and 10 minutes. The OTB infusion also significantly showed higher iron and cupper reducing powers thanthe TB infusion at the same time points. However, both samples demonstrated the same scavenging activity acrossall infusion times. These results suggest that the tea bags used with Twinings English Breakfast Tea may affectthe phenolic and flavonoid content released by the tea and subsequently influence the antioxidant ability of thetea as a reducing agent.
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Eucalyptus (Eucalyptus globulus Labill.) is an evergreen tree found worldwide. The aqueous ethanolic extract of the leaves (eucalyptus leaf extract; ELE) is used as a functional food, and its active constituents are generally polyphenols. Toxicity and mutagenicity of ELE have been previously assessed using rodents, and no adverse effects were observed. Although clinical trials of low-dose ELE ingestion have been conducted on humans, safety evaluation of high-dose ELE has not yet been conducted. We conducted an open-label clinical trial as a pilot study to assess the safety of excessive oral intake of ELE in Japanese adult men. A powdery preparation of ELE, commercially sold as Eucagrandin®, was prepared using spray drying method and contained approximately 80% ELE. Test capsules were packed with the powdery ELE. Six male subjects (aged 47.3 ± 12.4 years, mean ± SD) orally consumed 2,592 mg of ELE per day for 4 consecutive weeks. The subjects were examined at the start of the intervention, 4 weeks after the start, and 2 weeks after the last intake of test capsules. During the study period, 2 adverse events were reported. However, the causal relationship with the consumption of ELE was denied in both events because one event was attributed to a bruise and the other, which was elevated CPK levels, was due to excessive exercise. No unusual changes related to ELE consumption were observed in physical examination and during medical interviews. Further, blood and urine tests were normal during the trial period. These results demonstrated that oral consumption of ELE at the indicated dose is safe for humans.
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OBJECTIVE@#To investigate the phenolic composition, antioxidant properties, and hepatoprotective mechanisms of polyphenols from green tea extract (GTP) in carbon tetrachloride (CCl)-induced acute liver injury mouse model.@*METHODS@#High-performance liquid chromatography was used to analyze the chemical composition of the extract. Antioxidant activity of GTP was assessed by O, OH, DPPH, and ferric-reducing antioxidant power (FRAP) assay in vitro. Sixty Kunming mice were divided into 6 groups including control, model, low-, medium-, and high-doses GTP (200, 400, 800 mg/kg) and vitamin E (250 mg/kg) groups, 10 in each group. GTP and vitamin E were administered at a level of abovementioned doses twice per day for 7 days prior to exposure to a single injection of CCl. Hepatoprotective effects of GTP were evaluated in a CCl-induced mouse model of acute liver injury, using commercial enzyme linked immunosorbent assay kits, histopathological observation, terminal deoxynucleotidyl transferase-mediated dUTPNick-end labeling (TUNEL) assay and Western blot.@*RESULTS@#GTP contained 98.56 µg gallic acid equivalents per milligram extract total polyphenols, including epicatechingallate, epigallocatechin gallate, epicatechin, and epigallocatechin. Compared with the model group, low-, medium-, or high doses GTP significantly decreased serum levels of alanine aminotransferase and aspartate transaminase (P<0.01). Histopathological observation confirmed that pretreatment of GTP prevented swelling and necrosis in CCl-exposed hepatocytes. Hepatoprotective effects of low-, medium-, and high-dose GTP were associated with eliminating free radicals and improving superoxide dismutase, catalase, and glutathione peroxidase activity in the liver. Additionally, low-, medium-, and high-dose GTP decreased cell apoptosis in the CCl-exposed liver (P<0.01). Phosphorylated nuclear factor kappa-B (NF-κB), p53, Bcl-2 associated x protein/B-cell lymphoma/leukemia-2 gene, cytochrome C, and cleaved caspase-3 levels were downregulated compared with the model group (P<0.01).@*CONCLUSION@#GTP achieves hepatoprotective effects by improving hepatic antioxidant status and preventing cell apoptosis through caspase-3-dependent signaling pathways.
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Polyphenol oxidase(PPO) is an important antioxidant enzyme in plants. It has the functions of scavenging active oxygen and synthesizing phenols, lignin, and plant protection factors, and can enhance the plant's resistance to stress and resistance to pests and diseases. Our previous research found that Salvia miltiorrhiza PPO gene can positively regulate salvianolic acid B synthesis. In order to further explore the mechanism, a pGBKT7-PPO bait vector was constructed using the cloned S. miltiorrhiza polyphenol oxidase gene(SmPPO, GenBank accession number: KF712274.1), and verified that it had no self-activation and no toxicity. The titer of S. miltiorrhiza cDNA library constructed by our laboratory was 4.75 × 107 cfu·mL~(-1), which met the requirements for library construction. Through yeast two-hybrid test, 22 proteins that could interact with SmPPO were screened. Only yeast PAL1 and TAT interacted with SmPPO through yeast co-transformation verification. Further verification was performed by bimolecular fluorescence complementary detection(BiFC). Only TAT and SmPPO interacted, so it meant that TAT and SmPPO interacted. TAT and SmPPO were truncated according to the domain, respectively. The first 126 amino acids of SmPPO and tyrosine amino transferase(TAT) were obtained to interact on the cell membrane and chloroplast. SmPPO was obtained by subcellular localization test, which was mainly loca-lized on the nucleus and cell membrane; TAT was localized on the cell membrane. Real-time quantitative PCR results showed that the SmPPO gene was mainly expressed in roots and stems; the TAT gene was expressed in roots, and the expression level in stems and flowers was low. This article lays a solid foundation for the in-depth study of the molecular mechanism of the interaction of S. miltiorrhiza SmPPO and TAT to regulate the synthesis of phenolic substances.