Blueberry anthocyanins extract attenuates oxidative stress and angiogenesis on an in vitro high glucose-induced retinopathy model through the miR-33/GLCCI1 axis

Background:Diabetes retinopathy(DR)is a complication of diabetes that affects patients’vision.Previous studies have found blueberry anthocyanins extract(BAE)can inhibit the progression of DR,but its mechanism is not completely clear.Methods:To study the role of BAE in diabetes retinopathy,we treated human retinal endothelial cells(HRCECs)with 30 mM high glucose to simulate the microenvironment of diabetes retinopathy and used BAE to intervene the in vitro high glucose-induced retinopathy model.HRCEC cell viability and apoptosis rates were examined by Cell Counting Kit 8(CCK-8)assay and flow cytometry assay.The binding sites between miR-33 and glucocorticoid-induced transcript 1(GLCCI1)were assessed by luciferase reporter assay.Retinal neovascularization and oxidative stress contribute to diabetic retinopathy.The tubule formation assay was applied to detect the retinal neovascularization.The oxidative stress in the HRCECs was manifested by the reactive oxygen species(ROS)level,the malondialdehyde(MDA)level,and the superoxide dismutase(SOD)activity.Results:Compared with HRCECs cells cultured under normal conditions,high glucose(HG)can induce oxidative stress in HRCRCs,specifically manifested in the increase of ROS and MDA levels,and the decrease of SOD activity.BAE relieved the tubule formation in n the HRCEC.BAE also relieved the ROS and MDA levels and increased the SOD activity.Luciferase reporter assay revealed that GLCCI1 is a target molecule downstream of miR-33.In HRCEC,BAE significantly inhibited the expression of miR-33 induced by HG.miR-33 mimic inhibited the BAE’s effects on oxidative stress and angiogenesis in an in vitro high glucose-induced retinopathy model.Conclusion:BAE alleviated the oxidative stress and microangiogenesis of HRCEC by regulating the miR-33/GLCCI1 axis.

Metabolomic Analysis of the Anthocyanins Associated with Different Colors of Cymbidium goeringii in Guizhou, China

Cymbidium goeringii is an economically important ornamental plant,and flower color is one of the main features of C.goeringii that contributes to its high economic value.To clarify the molecular mechanisms underlying the role of anthocyanins in mediating differences in color among varieties,liquid chromatography–tandem mass spectrometry was used to perform anthocyanin-targeted metabolomics of seven C.goeringii varieties,including‘Jin Qian Yuan’(JQY),‘Jin Xiu Qian Yuan’(JXQY),‘Miao Jiang Su Die’(MJSD),‘Qian Ming Su’(QMS),‘Shi Chan’(SC),and‘Yang Ming Su’(YMS),as well as the C.goeringii.We detected 64 anthocyanins,including cyanidins,delphinidins,malvidins,pelargonidins,peonidins,petunidins,procyanidins,and flavonoids.We identified six shared differentially accumulated metabolites(DAMs),including cyanidin-3-O-rutinoside,delphinidin-3-Osophoroside,pelargonidin-3-O-rutinoside,peonidin-3-O-(6-O-malonyl-beta-D-glucoside),peonidin-3-Osophoroside,and chalcone.Most DAMs were enriched in the anthocyanin biosynthesis pathway.Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that the differentially expressed metabolites were significantly enriched in the anthocyanin biosynthesis pathway.Analysis of the content of differentially expressed metabolites indicated that peonidin-3-O-(6-O-malonyl-beta-D-glucoside)was the key metabolite underlying color differences among C.goeringii varieties.Procyanidin B2,pelargonidin-3-O-galactoside,and naringenin might also affect the color formation of JQY and QMS,SC,and MJSD,respectively.The results of this study shed light on the metabolic mechanism underlying flower color differences in C.goeringii at the molecular level.Our findings will aid future studies of the mechanism of flower color regulation in C.goeringii and have implications for the breeding of new varieties.

Quantification of Total Phenols, Total Flavonoids, Total Anthocyanins and Evaluation of Antioxidant and Antiradical Activities of Detarium Senegalense Extracts from Chad

The aim of the present work is to assess the value of Detarium Senegalense by determining the content of total phenols, total flavonoids and total anthocyanins, and by evaluating the free radical scavenging activity of Detarium Senegalense extracts. For this purpose, sequential extraction using solvents of increasing polarity was essential. The various extracts obtained underwent phytochemical and biochemical analyses. Phytochemical screening revealed the presence of flavonoids, alkaloids, tannins, polyphenols, anthocyanins and steroids/terpenes. Quantitative analysis of total polyphenols, total flavonoids and total anthocyanins yielded the following results: total flavonoids (0.803 ± 0029 mg EQ/100g P for acetone extract of roots and 0.871 ± 0.401 mg EQ/100g P for methanol extract of leaves);total polyphenols (23.298 ± 12.68 mg EAG/100g P for acetone extract of roots and 24.69 ± 0.49 401 mg EAG/100g P for methanol extract of leaves);total monomeric anthocyanins (44.697 ± 0.939 mg EC3G/100g P and 16.699 ± 0.193 mg EC3G/100g P respectively for acetone and methanol extracts of stem bark). DPPH free radical scavenging activity was 1.674 ± 0.023 mg/mL for the acetone extract and 0.934 ± 0.24 mg/mL for the methanol extract of roots. .

Transcriptomic Analysis Reveals the Formation Mechanism of Anthocyanins Light-Independent Synthesis in Chrysanthemum

Chrysanthemum×morifolium is a horticultural crop which plays a vital role in theflower industry with signifi-cant economic value and has a cultivation history of over three thousand years in China.The accumulation of anthocyanins is always affected by light.Here,we revealed that anthocyanin accumulation is highly dependent on light in‘2021135’genotype chrysanthemum,while it is light-independent in‘2001402’genotype chrysanthe-mum.However,no literature has been reported regarding the non-photosensitive chrysanthemum in anthocya-nins light-independent synthesis pathways.Through the phenotype analysis of 44 F1 generations,we found that light-independence is a dominant trait which can be stable inherited by progeny.The transcriptome of the rayflorets of‘2021135’and‘2001402’under light and bagging treatment were sequenced and analyzed.Based on weighted gene co-expression network analysis(WGCNA),K-means analysis,and Real-Time Quantitative Poly-merase Chain Reaction(RT-qPCR)analysis,16 genes were highly correlated with the anthocyanin content.The anthocyanin content of rayflorets treated with different light-quality conditions indicated that blue light signifi-cantly affected anthocyanin accumulations.Through Yeast one-hybrid analysis,CmBIC1.1 and CmBIC1.2 can directly regulate the anthocyanin structural gene CmCHS2.In our study,we revealed the important characteristics of light-independent anthocyanin synthesis in chrysanthemums and screened regulatory factors in light-depen-dent and light-independent anthocyanin synthesis pathways.The results laid the groundwork for subsequent ana-lysis of the molecular mechanism involved in the light-independent synthesis of anthocyanins in chrysanthemums.

Physiological Mechanism for Anthocyanins to Strengthen the Drought Tolerance of Plants

This paper summarized the possible physiological mechanism by which anthocyanins strengthen the tolerance of plants to drought. Drought stress can in-duce plant cel s to synthesize and accumulate anthocyanins. The photochemical properties, subcel ular accumulation sites and spatial distributions in plant organs and tissues of anthocyanins determine their function of strengthening plant tolerance, which is realized by three possible physiological mechanisms： （1） anthocyanins and their chelated metal ions can optimize the osmoregulation ability of the plant cel s by directly acting as the osmoregulation substances of the cel s, （2） anthocyanins with suitable spatial locations can reduce the photoinhibition of the plants under drought stresses, （3） anthocyanins can effectively maintain and improve the active oxygen-scavenging capacity of the plant cel s under drought conditions. Therein, that the anthocyanins enhance the antioxidant capacity of the plant cel s under drought stresses is probably the main reason for the anthocyanins to strengthen the drought tolerance of plants. This review could provide a reference for the mechanism re-search of the drought resistance and the breeding of the drought-resistant cultivars for the plants holding the ability to synthesize and accumulate anthocyanins.

Prediction Model of Secondary Substances in Anthocyanins Synthesis of Purple Corn

The aim of this study was to assay the polyphenols,flavonoid,polyphenol oxidase and phenylalnine ammonialyase which were relative to the anthocyanins synthesis of purple corn. The optimization of multiple linear regression model of anthocyanins synthesis was y=4.383 86-0.205 45x1＋5.479 638x2＋0.195 575x4. According to standard partial regression coefficient testing,the result indicated that polyphenols content was negatively correlated with anthocyanins and the relative influence to anthocyanins synthesis was-42.7%; flavonoid content and activity of polyphenol oxidase were positively correlated with anthocyanins of purple corn and the relative influence to anthocyanins synthesis were 71.45% and 73.32% respectively. There was no positive correlation between the activity of phenylalnine ammonialyase and anthocyanins of purple corn. The establishment of multiple linear regression model of anthocyanins synthesis was to provide theory foundation of producing anthocyanins in laboratory.

Effects of Cerium on Accumulation of Anthocyanins and Expression of Anthocyanin Biosynthetic Genes in Potato Cell Tissue Cultures

The effects of Ce （Ⅳ） on callus growth, anthocyanin content, and expression of anthocyanin biosynthetic genes in callus suspension cultures of Solanum tuberosum cv. Chieftain were studied by the measurement of fresh weight, spectrophotometric assays, and semiquantitative RT-PCR. The results indicate that 0.1 mmol·L^- 1 Ce （ Ⅳ ） can promote callus growth, increase the accumulation of anthocyanins, and enhance the expression of five anthocyanin biosynthetic genes （ CHS, F3H, F3＇5＇H, DFR, and 3 GT） most efficiently. At high concentrations of 1 mmol·L^- 1, Ce （Ⅳ） partially inhibits callus growth and at 2 mmol· L^-1 eventually lends to cell death. The results show that Ce（ Ⅳ ） can induce the expression of anthocyanin biosynthetic genes to produce and accumulate anthocyanins and increase the yield of anthocyanins.

Composition and antioxidant activity of anthocyanins isolated from Yunnan edible rose(An ning)

Edible roses(An ning)are a good source of anthocyanins and grown widely in Yunnan Province of China.In this study,the contents of anthocyanins and total phenol as well as the antioxidant activity of methanol extract from specific variety of rose were systematically investigated.The results showed that anthocyanins and total phenolic content of the petals were(353.56±2.50)mg cyanidin 3,5-diglucoside(Cy-3,5-diglu)equivalents and(2087.43±17.37)mg gallic acid equivalents(GAE)per 100 g fresh weight(FW),respectively.Totally,3 kinds of anthocyanins were detected and Cy-3,5-diglu was the predominant constituent which accounted for approximately 94.9%of total anthocyanins according to the analysis results of high performance liquid chromatography-photodiode array detection(HPLC-PAD).Data demonstrated that the extract from edible rose exhibited excellent ferric reducing capacity and free radical scavenging activity against both 2,2-diphenyl-1-picrylhydrazyl(DPPH)and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt(ABTS).The equivalents of anthocyanins from roses on DPPH,ABTS and ferric reducing ability were 2089,639 mg and 1400 mg GAE per 100 g FW,respectively.The high anthocyanins content and excellent antioxidant activity suggested that Yunnan edible roses could be applied in food industry as a good source of natural pigments.©2013 Beijing Academy of Food Sciences.Production and hosting by Elsevier B.V.All rights reserved.

Isolation and Purification of Anthocyanins from Blood Oranges by Column Chromatography

In this study, isolation and purification of anthocyanins from blood oranges by column chromatography were investigated, and then the anthocyanins of blood orange were identified. The behaviors of static adsorption and desorption, dynamic adsorption and desorption of 12 kinds of resins were compared. The results indicated that NKA-9 macroporous resin was optimum for isolation of blood orange anthocyanins, and the optimal elution reagent was 50% ethanol with citric acid （pH 2.5）. Toyopearl TSK HW-40S column was employed to separate and purify the anthocyanin extracts from blood orange. The best separation of Toyopearl TSK HW-40S column was obtained using a mobile phase of 35% methanol with 2% formic acid at a flow-rate of 0.6 mL min-~. Three kinds of anthocyanins were purified from blood orange. Then, the anthocyanins of blood orange were identified by HPLC-ESUMS analysis. The results showed that cyanidin-3-glucoside （35.2%） and cyaniding-3-（6＂-malonyl） glucoside （42.9%） were the major anthocyanins of blood orange. Furthermore, cyanidin-3-（3＂-malonyl） glucoside, cyanidin 3-（6＂-dioxalyl） glucoside and cyanidin-3-glucoside adduct：4-vinylcatechol were identified in blood orange. The combination of NKA-9 macroporous resin and Toyopearl TSK HW-40S column chromatography for isolation and purification of blood orange anthocyanins was an effective method, and HPLC-ESI/MS analysis was a convenient, rapid and effective method for identification of anthocyanins from blood orange.

Thermal Degradation Kinetics of Three Kinds of Representative Anthocyanins Obtained from Blood Orange

Cyanidin-3-glucoside, cyanidin-3-（6′′-malonyl）-glucoside, and cyanidin-3-glucoside-derived pyranoanthocyanins which were three major anthocyanins of blood orange, were obtained using a Toyopearl TSK HW-40S column chromatography. Then, thermal degradation kinetics of the three major anthocyanins was studied at selected temperatures （70, 80, and 90°C）. Degradation parameters such as k and t1/2 values were determined. The activation energy （Ea） values for cyanidin- 3-glucoside, cyanidin-3-（6′′-malonyl）-glucoside and cyanidin-3-glucoside-derived pyranoanthocyanin were 75.4, 79.5, and 81.7 kJ mol-1, respectively. Ea values suggested that cyanidin-3-glucoside-derived pyranoanthocyanin had the highest stability, followed by cyanidin-3-（6′′-malonyl）-glucoside and cyanidin-3-glucoside. However, t1/2 values indicated cyanidin- 3-glucoside-derived pyranoanthocyanin degraded faster than cyanidin-3-（6′′-malonyl）-glucoside and cyanidin-3-glucoside at selected temperature.

Thermal Degradation Kinetics of Anthocyanins and Visual Color of Blood Orange Juice

Thermal degradation kinetics of anthocyanins and visual color （Hunter α value） of blood orange juice were studied at selected temperatures （70-90℃）. Results indicated that both the thermal degradation of anthocyanin and visual color all followed first-order reaction kinetics, and they could be expressed by Arrhenius equation. The activation energy values for the anthocyanins degradation and visual color degradation were 55.81 and 47.51 kJ tool-1, respectively. The linear relationship between visual color and anthocyanin content was obtained. Furthermore, during thermal processing of blood orange juice, the formulas about the linear relationships showed no significant difference at selected temperatures. So, the relationships between visual color and anthocyanins content during thermal processing at selected temperatures could be described by the same equation： α＊/αo＇=0.559（C/Co）＋0.43. It might be inferred that visual color measured instantaneously by tristimulus colorimeters for on-line quality control, could be used to predict the anthocyanins degradation during thermal processing of blood orange juice.

<i>In Silico</i>Analysis of a MRP Transporter Gene Reveals Its Possible Role in Anthocyanins or Flavonoids Transport in <i>Oryze sativa</i>

There are many studies on enzymatic pathways of anthocyanin biosynthesis, but little is known about the anthocyanins transport in Oryze sativa. In silico analysis, the OsMRP15 (LOC_Os06g06440), an orthologous gene of mazie anthocyanin transporter ZmMRP3, has been identified in rice. The OsMRP15 contained a 4425bp open reading frame (ORF) encoding a 1475 amino acid protein, belonging to a MRP subfamily of ABC transporters, and has a high sequence identity, very similar protein structure, and the same arrangement of domains to ZmMRP3, but the genomic structure of OsMRP15 was significant difference with ZmMRP3. The prediction promoter of OsMRP15 has many presumed anthocyanin regulatory sites. The phylogenetic analysis of MRPs in rice, mazie and Arabidopsis showed that OsMRP15 and ZmMRP3 belonged to the same subbranch. The expression pattern indicated that OsMRP15 was co-expression with two anthocyanin transcription factors. These analysis results implied that as an ortholog of ZmMRP3, the function of OsMRP15 was possibly as a membrane-bound transporter required for vacuolar uptake of anthocyanins in rice.

Extraction Characteristics of Anthocyanins from Preliminarily-processed Products of Purple Sweet Potatoes during the Gelatinization Process

Objective] This study aimed to investigate the method for efficient utilization and development of purple sweet potatoes. [Method] Purple sweet potatoes were dried at two specific temperatures and prepared into preliminarily-processed products for gelatinization simulation to analyze the extraction amount of anthocyanins from gelatinized samples at different gelatinization stages. [Result] During the gelatinization process, the extraction rate of anthocyanins from purple sweet potato samples reached the highest as the temperature rised from 90 ℃ to 95 ℃,and the extraction amount of anthocyanins reached the maximum at 15 min postheat preservation at 95 ℃. Purple sweet potato samples dried at 60 ℃ exhibited larger retention amount, larger maximum extraction amount and higher maximum extraction rate of anthocyanins compared with those dried at 110 ℃. [Conclusion] Drying at low temperatures and appropriately shortening the initial gelatinization stage below 90 ℃ is conducive to the retention and extraction of anthocyanins from purple sweet potatoes.

Density Functional Theoretical Study on Antioxidant Activity of Four Anthocyanins from Purple Sweet Potato

This study aimed to analyze the antioxidant mechanism of four different anthocyanins in purple sweet potato. The DPPH test and the lipid peroxidation inhibition ability test were used to analyze the in vitro antioxidant activity of four anthocyanins, and then the optimal structure, bond dissociation energy(BDE) and ionization potential(IP) of the anthocyanins and their simplified molecular models were analyzed by density functional theoretical study, and the antioxidant mechanisms were explored. The results showed that OH-4′ phenolic hydroxyl group of the four anthocyanins has the highest activity, and its bond dissociation energy is less than that of resveratrol. The theoretical calculation results of antioxidant activity are consistent with the results of in vitro antioxidant tests. The results showed that anthocyanins of purple sweet potato have good antioxidant activity, and the DFT method provides a powerful theoretical basis for the development of anthocyanin antioxidant activity.

Chemical Composition, Phenolics, Anthocyanins Concentration and Antioxidant Activity of Ten Wild Edible Plants

Plants were collected and prepared for chemical analysis, total phenolics, anthocyanins concentrations, and free radical scavenging activity. Results showed that, protein concentration of Malva parviflora (22.9%) was the highest among the plants. Ruta chalepensis had high levels of fat and carbohydrates (4.2% and 51.7%, respectively), but had the lowest level of ash (8.7%). Mineral concentrations varied and found to have appreciable amounts of Ca, Na, K, Cu, Fe, Mg, Mn, Zn and P. Total phenolic ranged from 163.1 (Tetragonolobus palaestinus) to 1328.8 mg GAE/100g (Ruta chalepensis). Anthocyanins ranged between 18.1 (Gundelia tournefortii) and 100.1 mg/100g (Rumex acetosella). These plants differed in free radical scavenging activity. It was concluded that these plants could be considered as natural sources for antioxidants and valuable natural resources as a new addition to the diet of inhabitants.

Variation of Polyphenols, Anthocyanins and Antioxidant Power in the Strawberry Grape (<i>Vitis labrusca</i>) after Simulated Gastro-Intestinal Transit and Evaluation of <i>in Vitro</i>Antimicrobial Activity

The influence of a simulated digestive process on some biochemical and biological aspects of strawberry grape (Vitis labrusca) was investigated. The amount of total polyphenols and anthocyanins as well as the antioxidant power were evaluated. Results evidenced that the simulated gastrointestinal transit caused a decrease of the polyphenols content and total anthocyanins;these last, however, were more resistant than polyphenols, decreasing only of 50% respect to the initial value (31.50 μg/ml of extract). The extract exhibited an excellent antioxidant power (EC50 3.8 mg/ml), which decreased of about four times after the simulated gastrointestinal transit. The antimicrobial activity of the extract, evaluated against three Escherichia coli, Pseudomonas aeruginosa and Bacillus cereus pathogen strains was enhanced by the simulated digestion, with an increase of the inhibition halo.

Extraction and Stability of Anthocyanins Present in the Skin of the Dragon Fruit (<i>Hylocereus undatus</i>)

The extraction of anthocyanins present in the skin of the dragon fruit was performed using trifluoroacetic acid (TFA) plus a mixture of methanol, acetic acid and water;the anthocyanins were then purified with a LC-18 cartridge, using methanol acidified with TFA as eluent, reaching concentrations of 44.3865 ± 1.3125 mg/100g of sample. The extracts were put through stability tests under different storage conditions, modifying the pH of the extracts (pH of 1, 4 and 6), the temperature (4℃, 25℃ and 68℃) and the absence and presence of light for a time period of 4 days;the tests indiated that anthocyanins remain more stable at a temperature of 4℃?with a pH of 4 in the absence of light, retaining up to 80% of the pigment. Three anthocyanins were partially identified in the extracts by high performance liquid chromatography (HPLC);they were: cyanidin 3-O-glucoside, cyanidin 3,5 O-glucoside and pelargonidin 3,5 O-glucoside.

Effect of Light on Stability of Anthocyanins in Ethanolic Extracts of Rubus fruticosus

Blackberry (Rubus fructicosus) is one of the fruit with the highest concentration of anthocyanins;however, its use is limited for making jams, jellies and liqueur, and recently, fruit concentrates in combination with pomegranate, blueberry and grape. One of the main problems with these pigments is their poor stability in solution, mainly in beverages as liqueur and juices, which depends on factors such as chemical structure, pH, temperature, light, water activity, and presence of oxygen. The effect of light on total monomeric anthocyanins content as well as the degradation rates and browning of two blackberry ethanolic extracts is to establish the conditions for storage of liqueur without adding artificial food coloring. The initial content of anthocyanins on extract without storage was 106 mg?l-1. The study assessed the light irradiation effect on the anthocyanins of blackberry ethanolic extract. The anthocyanins degradation followed the second order reaction kinetics with respect to illuminance of the light source. The t1/2 value at high illuminance (3968.30 lx) was 28.20 hours.

Temperature and Acidified Solvent Effect on Total Anthocyanins and RP-HPLC Phenolic Acids Determination in Selected Spices

Total anthocyanins of spices (Syzygium aromaticum L., Coriadrum sativum L., Cuminum cyminum L., Zingiber officinale Rosc., Elettaria cardamomum, Curcuma longa, Rhus coriaria L., Cinnamomum zeylanicum Blume, Foeniculum vulgare Mill and Laurus nobilis L.) were determined using acidified (1% HCl) solvents (methanol, ethanol and acetone) at three temperatures (20℃, 40℃ and 60℃). Also phenolic acids were separated and identified by RP-HPLC. Results showed that sumac and cinnamon had the highest levels of anthocyanins, while for the acetone the cinnamon indicated the highest amount of anthocyanins when methanol and ethanol were used as extracting solvents at 20℃. At 40℃ using ethanol, sumac showed the highest level of anthocyanins whereas acetone solvent yielded the highest anthocyanin contents for cinnamon. At 60℃, cinnamon showed the highest level of anthocyanins when methanol and acetone were the solvents, while sumac had the highest anthocyanins level using ethanol as solvent. HPLC results showed ten phenolic acids found in those spices and varied in their concentrations. Gallic acid had the highest level (1642.3 mg/100g) (cloves). Gentisic acid had the lowest level (1.2 mg/100g) in ginger. Also sumac showed the highest level of chlorogenic acid (1528.7 mg/100g). Some acids were not found in some spices, for instance, benzoic acid was not found in coriander, cumin, ginger, green cardamom, cinnamon and sweet laurel.

Eco-Friendly pH Indicator Based on Natural Anthocyanins from Lycium ruthenicum

An eco-friendly and visual pH indicator was developed based on cotton fabrics dyed with natural Lycium ruthenicum extract(LRE),whose color changed from red to purple,blue and green at pH of 2-10.Three methods for dyeing cotton fabrics with LRE were attempted,namely pre-mordanting,simultaneous mordanting,and post-mordanting methods with different dyeing temperatures,dyeing times and dyebath pH values.The cotton fabrics exhibited the highest K/S value when they were dyed under a dyebath of pH 6 at 20℃for 90 min in the case of the simultaneous mordanting method.Meanwhile,the dyed cotton fabrics also showed reversible pH-dependent color changes.The developed flexible pH indicator based on renewable natural materials is suitable for multiple applications in environmental monitoring and smart textiles.