Isolation of High-Purity Betanin from Red Beet and Elucidation of Its Antioxidant Activity against Peroxynitrite: An in vitro Study.

Reactive oxygen species and reactive nitrogen species (RNS) are damaging for many biomolecules. Peroxynitrite (ONOO-) is the most toxic molecular species among RNS. Betalains are known to possess ONOO- scavenging ability. Betanin, a betalain isolated from red beet, possesses antioxidant, anti-inflammatory, and antitumor activities; however, detailed studies of this isolated pigment have not been conducted, owing to its instability under physiological conditions. This study aimed to isolate highly purified betanin from red beetroots using an improved purification method involving deproteinization and citric acid co-precipitation and evaluated its antioxidant activities. The purified betanin thus obtained had a significantly lower isobetanin content than the commercially available betanin dyes. The antioxidant activity of purified betanin examined in the 2,2-diphenyl-1-picrylhydrazyl assay, the direct ONOO- reaction, ONOO--dependent DNA damage, and lipid peroxidation reactions revealed that betanin possessed higher antioxidant capacity than general antioxidants such as ascorbic acid and quercetin. Furthermore, betanin showed indirect and direct cytoprotective effects against H2O2 and ONOO- cytotoxicity, respectively, in cultured mouse fibroblasts. To the best of our knowledge, this is the first study to demonstrate the cytoprotective effects of betanin against ONOO- toxicity. The highly purified betanin obtained in this study will aid in further exploring its physiological functions.

Effects of heat degradation of betanin in red beetroot (Beta vulgaris L.) on biological activity and antioxidant capacity.

Betanin is a red pigment of red beetroot (Beta vulgaris L.), providing the beneficial effects to maintain human health. Betanin is involved in the characteristic red color of red beetroot, and used as an edible dye. Betanin is known to be a highly unstable pigment, and water solutions of betanin are nearly fully degraded after heating at 99°C for 60 min in the experimental conditions of this study. The present study investigated the effects of red beetroot juice (RBJ) and betanin on immune cells, and found that stimulation with RBJ and betanin induces interleukin (IL)-1ß, IL-8, and IL-10 mRNA in a human monocyte derived cell line, THP-1 cells. This mRNA induction after stimulation with RBJ and betanin was not significantly changed after heat treatment when attempting to induce degradation of the betanin. Following these results, the effects of heat degradation of betanin on the inhibition of lipopolysaccharide (LPS) induced nitric oxide (NO) production in RAW264 cells and the antioxidant capacity were investigated. The results showed that the inhibition activity of RBJ and betanin with the LPS induced NO production is not altered after heat degradation of betanin. In addition, the results of FRAP (ferric reducing antioxidant power) and DPPH (1,1-Diphenyl-2-picrylhydrazyl) assays indicate that a not inconsiderable degree of the antioxidant capacity of RBJ and betanin remained after heat degradation of betanin. These results suggest that it is important to consider the effects of degradation products of betanin in the evaluation of the beneficial effects of red beetroot on health.

Effect of two-week red beetroot juice consumption on modulation of gut microbiota in healthy human volunteers - A pilot study.

With very little research exploring intestinal effects of red beetroot consumption, the present pilot study investigated gut microbial changes following red beetroot consumption, via a 14-day intervention trial in healthy adults. Compared to baseline, the study demonstrates transient changes in abundance of some taxa e.g., Romboutsia and Christensenella, after different days of intervention (p < 0.05). Enrichment of Akkermansia muciniphila and decrease of Bacteroides fragilis (p < 0.05) were observed after 3 days of juice consumption, followed by restoration in abundance after 14 days. With native betacyanins and catabolites detected in stool after juice consumption, betacyanins were found to correlate positively with Bifidobacterium and Coprococcus, and inversely with Ruminococcus (p < 0.1), potentiating a significant rise in (iso)butyric acid content (172.7 ± 30.9 µmol/g stool). Study findings indicate the potential of red beetroot to influence gut microbial populations and catabolites associated with these changes, emphasizing the potential benefit of red beetroot on intestinal as well as systemic health.

Inhibition of Proliferation in Ovarian Cancer Cell Line (PA-1) by the Action of Green Compound "Betanin".

Ovarian carcinoma has a cure rate of 30% which makes it deadlier than any other disease. There are a number of genetic and epigenetic changes that lead to ovarian carcinoma cell transformation. Chemoprevention of cancer through application of natural compounds is the need of present generation as other methods are rigorous and have many side effects. Betanin, a compound from Beta vulgaris extract is used in present study to check its potential for inhibition of (PA-1) cancer cell proliferation. Determination of IC50 values through MTT assay was carried out, in addition measurement of mitochondrial membrane potential (MMP), effect of reactive oxygen species (ROS) generation, and induction of apoptosis in ovarian cancer cells through betanin was also observed. Results have shown betanin as a potential candidate for inhibition of ovarian cancer cell proliferation and it can be taken up as a serious compound for further studies for its application in cancer cure.

The impacts of peel inclusion and fermentation temperature on the bioactive compounds, antioxidant activity, and sensory properties of dragon fruit wines.

BACKGROUND: Even though dragon fruit peel has more health benefits than its pulp, it is considered to be waste after fruit processing. In this study, dragon fruit peel was explored as an additional ingredient in winemaking. METHODS: The contents of total phenolic compounds, betacyanins and ascorbic acid, the antioxidant capacity by DPPHz method, and the level of consumer acceptability of the wines prepared from two species with and without peel were compared. RESULTS: The wines of the H. polyrhizus species contained much higher total phenolics (1.4-1.6 times), betacyanins (75-81 times), and vitamin C (2.8-3.8 times), as well as higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (1.2 times) than the wines of H. undatus. Although the fermentation process led to the degradation of betacyanins, it increased the total phenolics (up to 17%), vitamin C (up to 24%), and DPPHz scavenging activity (up to 19%) as compared with the control when fermentation was conducted at 25°C, which was found as the most suitable fermentation temperature to produce wines with the highest quality in terms of antioxidant content and activity. At this temperature, peel inclusion also increased these values in the wine. In addition, the sensory properties in terms of appearance and aroma could be improved in the final fermented product. CONCLUSIONS: H. polyrhizus wine might have higher antioxidant properties than H. undatus wine. With a suitable fermentation temperature, peel inclusion as an ingredient could enhance these characteristics of the wine without compromising its sensory quality. This study may provide insights into fruit winemaking with higher health benefits.

Robust W/O/W Emulsion Stabilized by Genipin-Cross-Linked Sugar Beet Pectin-Bovine Serum Albumin Nanoparticles: Co-encapsulation of Betanin and Curcumin.

Betanin and curcumin hold promise as natural colorants and antioxidants for food purposes due to their anti-hypertensive, anti-inflammation, and anti-tumor effects. However, the thermal stability and bioavailability of betanin and curcumin still need improvement. Here, we fabricated sugar beet pectin-bovine serum albumin nanoparticles (SBNPs) with a mean particle size of 180 ± 5.2 nm through a genipin cross-linking strategy to stabilize a type of Pickering water-in-oil-in-water (W/O/W) emulsion and co-encapsulated betanin and curcumin. First, the W1/O emulsion was homogenized with gelatin (the gelling agent) in the water phase and polyglycerol polyricinoleate (a lipophilic surfactant) in the oil phase. Later, W1/O was homogenized with another water phase containing SBNPs. The microstructure of the emulsion was regulated by the particle concentration (c) and W1/O volume fraction (Φ), especially the gel-like high internal phase emulsions were formed at the Φ up to 70%. In this case, betanin was encapsulated in the internal water phase (encapsulation efficiency = 65.3%), whereas curcumin was in the medium-chain triglyceride (encapsulation efficiency = 84.1%). Meanwhile, the shelf stability of betanin and curcumin was improved. Furthermore, the stability of bioactive compounds was potentiated by an emulsion gel in simulated gastrointestinal digestion, resulting in higher bioaccessibility. The aforementioned results suggest that SBNP-stabilized Pickering W/O/W emulsions could be a potential alternative to co-encapsulate betanin and curcumin with enhancement of shelf stability and bioaccessibility.

Betacyanin-inhibited biofilm formation of co-culture of Staphylococcus aureus and Pseudomonas aeruginosa on different polymer surfaces.

Staphylococcus aureus and Pseudomonas aeruginosa are bacteria that cause biofilm-associated infections. The aim of this study was to determine the activity of combined betacyanin fractions from Amaranthus dubius (red spinach) and Hylocereus polyrhizus (red pitahaya) against biofilms formed by co-culture of S. aureus and P. aeruginosa on different polymer surfaces. Various formulations containing different concentrations of the betacyanin fractions were investigated for biofilm-inhibiting activity on polystyrene surfaces using crystal violet assay and scanning electron microscopy. A combination of each betacyanin fraction (0.625 mg mL-1) reduced biofilm formation of five S. aureus strains and four P. aeruginosa strains from optical density values of 1.24-3.84 and 1.25-3.52 to 0.81-2.63 and 0.80-1.71, respectively. These combined fractions also significantly inhibited dual-species biofilms by 2.30 and reduced 1.0-1.3 log CFU cm-2 bacterial attachment on polymer surfaces such as polyvinyl chloride, polyethylene, polypropylene and silicone rubber. This study demonstrated an increase in biofilm-inhibiting activity against biofilms formed by two species using combined fractions than that by using single fractions. Betacyanins found in different plants could collectively be used to potentially decrease the risk of biofilm-associated infections caused by these bacteria on hydrophobic polymers.

Betanin purification from red beetroots and evaluation of its anti-oxidant and anti-inflammatory activity on LPS-activated microglial cells.

Microglial activation can release free radicals and various pro-inflammatory cytokines, which implicates the progress of a neurodegenerative disease. Therefore suppression of microglial activation can be an appropriate strategy for combating neurodegenerative diseases. Betanin is a red food dye that acts as free radical scavenger and can be a promising candidate for this purpose. In this study, purification of betanin from red beetroots was carried out by normal phase colum chromatography, yielding 500 mg of betanin from 100 g of red beetroot. The purified betanin was evaluated by TLC, UV-visible, HPLC, ESI-MASS, FT-IR spectroscopy. Investigation on the inhibitory effect of betanin on activated microglia was performed using primary microglial culture. The results showed that betanin significantly inhibited lipopolysaccharide induced microglial function including the production of nitric oxide free radicals, reactive oxygen species, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1 beta (IL-1ß). Moreover, betanin modulated mitochondrial membrane potential, lysosomal membrane permeabilization and adenosine triphosphate. We further investigated the interaction of betanin with TNF-α, IL-6 and Nitric oxide synthase (iNOS or NOS2) using in silico molecular docking analysis. The docking results demonstrated that betanin have significant negative binding energy against active sites of TNF-α, IL-6 and iNOS.

Effect of Betanin, a Beetroot Component, on Vascular Tone in Isolated Porcine Arteries.

BACKGROUND: Beetroot has attracted much attention because of its blood pressure-lowering properties. Although beetroot contains various nutritional compounds, including inorganic nitrate, some of their physiological properties are not fully understood. In this study, we examined whether betanin, a beetroot component, has a regulatory effect on vascular tone. METHODS: Mechanical responses of isolated porcine coronary, mesenteric, and pulmonary arteries were assessed by organ chamber technique. In some cases, the vascular reactivity was observed in the presence of a physiological concentration of betanin (10 µM). RESULTS: Betanin did not induce vasorelaxation at physiological concentrations both in endothelium-intact and -denuded coronary, mesenteric, and pulmonary arteries. The endothelium-dependent agonists, bradykinin and A23187 induced vasorelaxation of endothelium-intact coronary arteries, both of which were not affected by exposure to betanin. Likewise, endothelium-independent vasorelaxation induced by sodium nitrite and sodium nitroprusside was also not affected by the presence of betanin. In addition, exposure of endothelium-intact coronary arteries to betanin did not attenuate prostaglandin F2α- and endothelin-1-induced vasocontraction. CONCLUSIONS: These findings suggest that betanin does not have a vasorelaxant activity. It is unlikely that betanin is a component directly responsible for the beetroot-induced acute blood pressure-lowering effect in a nitrate-independent manner.

Impact of betanin against paracetamol and diclofenac induced hepato-renal damage in rats.

Context: Paracetamol (PAR) and diclofenac (DF) are the most popular consumed analgesics and anti-inflammatory medications.Objective: This study aimed to explore the protective effect of betanin (Bet) against PAR or DF induced hepato-renal damage in rats.Methods: Rats were randomly divided into five groups: Normal control (NC) group rats were given saline only. PAR group rats received PAR (400 mg/kg). PAR/Bet treated group rats administered PAR (400 mg/kg) plus Bet (25 mg/kg). DF group rats received DF (10 mg/kg). DF/Bet treated group rats administered DF (10 mg/kg) plus Bet (25 mg/kg). All drugs were given by gavage for 28 consecutive days.Results: PAR and DF administration in high dose and long-time induced liver and kidney injury, disrupted serum lipid profile, enhanced serum levels of inflammatory and oxidative stress markers, triggered DNA fragmentation and caused drastic changes in the histopathological pictures of the two organs. Bet supplementation succeeded to ameliorate most of the biochemical changes and protected DNA from damage as obtained from comet assay. Histological features in H&E taken to different groups also mirrors this findings.Conclusion: Bet exerted a potential anti-inflammatory and antioxidant effect against hepato-renal damage induced by PAR or DF overconsumption.

Protective effects of betanin against paracetamol and diclofenac induced neurotoxicity and endocrine disruption in rats.

Context: Overconsumption of paracetamol (PAR) and diclofenac (DF) have been reported to induce neurotoxicity and endocrine disruption. Objective: The current study was designed to explore the protective potential of betanin against PAR and DF inducing neurotoxicity and endocrine disruption in a rat model. Material and Methods: Forty rats were equally divided into five groups: group I served as control, group II received PAR (400 mg/kg), group III received PAR plus betanin (25 mg/kg), group IV received DF (10 mg/kg) and group V received DF plus betanin orally for 28 consecutive days. Thyroid axis hormones, sex hormone, neurotransmitters, paraoxonase-1, hemeoxygenase-1 and nuclear factor-2 were measured by ELISA. While, the oxidative stress markers were colorimetrically estimated. Moreover, DNA damage and histopathological picture of the brains were investigated. Results: A marked reduction in thyroid axis hormones, brain neurotransmitters and serum testosterone as well as enhanced oxidative stress and brain DNA damage accompanied by drastic changes in the brain histopathological picture were recorded in the challenged PAR and DF groups. Betanin supplementation ameliorated most of the biochemical and histopathological changes induced by PAR or DF. Conclusion: The study suggests betanin of potential protective effects against neurotoxicity and endocrine disruption induced by PAR and DF overconsumption.

Betanin assisted synthesis of betanin@silver nanoparticles and their enhanced adsorption and biological activities.

Natural red purple dye, Betanin, was extracted from the beetroot, purified by aqueous two- phase extraction and gel permeation column chromatography, and used as a reducing agent for the synthesis of silver-betanin core-shell triangular nanodisks for the first time. Spectroscopic data show that the nanoparticle structure is core@shell like with Ag as core and betanin as shell. Langmuir monolayer model (qm = 32.4 mg/g, RL = 0.99 and R2 = 0.997) was the best fit adsorption isotherm for the dye removal. Adsorption kinetics is well explained by pseudo-second-order equation. Gibbs free energy (ΔG0 = -2.59 kJ mol-1), enthalpy (ΔH0 = 18.05 kJ mol-1), and entropy (ΔS0 = 68.92 J/K/mol) were calculated and discussed. The antibacterial activity of betanin-AgNPs were determined against Escherichia coli MTCC-450 (E. coli) and Staphylococcus aureus MTCC-3160 (S. aureus) bacteria.

Extension of life-span using a RNAi model and in vivo antioxidant effect of Opuntia fruit extracts and pure betalains in Caenorhabditis elegans.

Betalains are nitrogenous plant pigments known for their high antioxidant capacity. For the first time, this antioxidant nature has been studied in an in vivo system using the model organism Caenorhabditis elegans. The oxidative stress caused in the fluorescent strain TJ375 (hsp-16.2::GFP) was reversed by the presence of both natural and semi-synthetic betalains, with an ED50 value around 25 µM for betacyanins and up to 10 µM for betaxanthins, with indicaxanthin, the major pigments in prickly pear fruits, as the most effective betalain. The effect of model betalains on the lifespan of the wild-type N2 strain was carefully studied using the automatic platform "Lifespan Machine". In a search for different approaches to suppress progeny, pop-1 RNAi was used to avoid FUdR use. The presence of betalains in the medium, both as pure compounds and as enriched Opuntia extracts significantly increased the lifespan of C. elegans.

Biofilm inhibiting activity of betacyanins from red pitahaya (Hylocereus polyrhizus) and red spinach (Amaranthus dubius) against Staphylococcus aureus and Pseudomonas aeruginosa biofilms.

AIMS: To investigate the biofilm inhibitory activity of betacyanins from red pitahaya (Hylocereus polyrhizus) and red spinach (Amaranthus dubius) against Staphylococcus aureus and Pseudomonas aeruginosa biofilms. METHODS AND RESULTS: The pulp of red pitahaya and the leaves of red spinach were extracted using methanol followed by subfractionation to obtain betacyanin fraction. The anti-biofilm activity was examined using broth microdilution assay on polystyrene surfaces and expressed as minimum biofilm inhibitory concentration (MBIC). The betacyanin fraction from red spinach showed better anti-biofilm activity (MBIC: 0·313-1·25 mg ml-1 ) against five Staph. aureus strains while the betacyanin fraction from red pitahaya showed better anti-biofilm activity (MBIC: 0·313-0·625 mg ml-1 ) against four P. aeruginosa strains. Both betacyanin fraction significantly reduced hydrophobicity of Staph. aureus and P. aeruginosa strains. Numbers of Staph. aureus and P. aeruginosa attached to polystyrene were also reduced without affecting their cell viability. CONCLUSION: Betacyanins can act as anti-biofilm agents against the initial step of biofilm formation, particularly on a hydrophobic surface like polystyrene. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first to investigate the use of betacyanin as a biofilm inhibitory agent. Betacyanin could potentially be used to reduce the risk of biofilm-associated infections.

Enhancing the antimicrobial and antifungal activities of a coloring extract agent rich in betacyanins obtained from Gomphrena globosa L. flowers.

Although less explored than beetroot (Beta vulgaris L.), the flowers of Gomphrena globosa L. are a very suitable source of betacyanins with strong pigmentation features, together with many other desirable bioactive properties. Thus, the aim of this study was to enhance the antimicrobial and antifungal activities of a pigmented extract obtained from G. globosa flowers by ultrasound assisted extraction (UAE). The procedure was supported with the application of the response surface methodology, a robust optimization technique that allows to study jointly the effects of several variables and responses. To enhance the antimicrobial (Bacillus cereus, Listeria monocytogenes, Escherichia coli and Salmonella typhimurium) and antifungal (Aspergillus flavus, Aspergillus niger, Penicillium ochrochloron and Penicillium verrucosum) activities, the responses were evaluated in terms of the concentrations needed to obtain minimum inhibitory (MIC), minimum bactericidal (MBC) and minimum fungicidal (MFC) concentrations. It was found that the optimal UAE conditions were 10.8 min, 410.5 W, 57.8% of ethanol, and 5 g L-1 of the solid-liquid ratio providing the following response values: (1) from the studied species of bacteria, the MIC ranged from ∼0.15 to 0.35 g L-1 and the MBC ranges were ∼0.30 to 0.65 g L-1; and (2) from the studied fungus species, the MIC ranged from ∼0.20 to 0.30 g L-1 and the MFC ranges were ∼0.40 to 0.65 g L-1. The antibacterial activity dose levels were lower than the antifungal ones. In conclusion, the results obtained in this study highlight extracts from G. globosa flowers as natural sources of betacyanins with application as food colorants with important antimicrobial and antifungal activities.

Liquid chromatographic techniques in betacyanin isomers separation from Gomphrena globosa L. flowers for the determination of their antimicrobial activities.

High-speed counter-current chromatography (HSCCC) combined with HPLC were established to purify, for the first time, betacyanin isomers from Gomphrena globosa L. flowers. The mixture of tert-butyl methyl ether - butanol - methanol - water TBME-BuOH-MeOH-H2O (1.0% heptafluorobutyric acid HFBA) 2:2:1:5 (v/v/v/v) in reversed phase mode was used for preliminary purification of gomphrenin derivatives. Then, semiprep-HPLC was used to obtained ten pure betacyanins: gomphrenins II and III with their cis-isomers as well as sinapoyl-gomphrenin I and its corresponding diastereomers. This is the first study on separation of naturally coexisting acylated gomphrenin isomers. The isolated compounds showed a broad antimicrobial spectrum (in the range of 0.19-1.5 mg mL-1) by inhibiting the growth of all of food-borne pathogens tested. The antimicrobial activity of acylated betacyanins was higher than that of non-acylated gomphrenin I/isogomphrenin I. Therefore, the acylation with phenolic acids increases the activity of betacyanins against fungi and bacteria, suggesting a synergetic effect of the acylated moieties and the pigment chromophoric system on the activity, which was evaluated for the first time.

Beetroot and Sodium Nitrate Ameliorate Cardiometabolic Changes in Diet-Induced Obese Hypertensive Rats.

SCOPE: Dietary intake of beetroot by humans reduces blood pressure but whether this is caused by nitrate or betanin is not well-defined; neither are effects on other signs of metabolic syndrome. METHODS AND RESULTS: Rats fed a high-carbohydrate, high-fat diet (H) for 16 weeks developed abdominal obesity, hypertension, altered cardiovascular and liver structure and function, and impaired glucose tolerance compared to rats fed a corn starch diet (C). H rats treated with ∼16 mg/kg/day of nitrate either from beetroot juice (H+B) or sodium nitrate (H+N) for the last 8 weeks reduced systolic blood pressure by ∼25 mmHg, improved cardiac structure and function, plasma lipid profile and plasma markers of liver function, reduced inflammatory cell infiltration in heart and liver and decreased left ventricular fibrosis. In the left ventricle, H rats increased mRNA expression of connective tissue growth factor (CTGF), monocyte chemoattractant protein 1 (MCP-1), matrix metalloproteinase-2 (MMP-2), and adenosine monophosphate-activated protein kinase-alpha (AMPK-α) and decreased mRNA expression of peroxisome proliferator-activated receptor-alpha (PPAR-α); both beetroot and sodium nitrate diet-fed rats decreased CTGF threefold, MCP-1, and MMP-2 twofold, and doubled PPAR-α mRNA expression in left ventricular tissue. CONCLUSION: The similar functional and molecular responses to beetroot and sodium nitrate indicate that the nitrate content of beetroot reduced inflammation and improved cardiovascular, liver, and metabolic function in rats with metabolic syndrome, rather than betanin.

Betanin, isolated from fruits of Opuntia elatior Mill attenuates renal fibrosis in diabetic rats through regulating oxidative stress and TGF-ß pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The fruits of Opuntia elatior Mill are being used traditionally in different disease condition like diabetes, obesity, asthma, inflammatory disorders, and anemia. Betanin, a compound isolated from fruits of Opuntia elatior Mill has potent anti-oxidative and anti-inflammatory activity. Recent study from our lab indicated the protective effect of betanin against high glucose induced rat renal epithelial cell fibrosis and matrix accumulation, major features of diabetic nephropathy (DN). However the molecular mechanism of betanin in DN has not yet been fully elucidated. AIM OF THE STUDY: The aim of the present study was to further investigate the anti-fibrotic mechanisms of betanin against streptozotocin (STZ) induced DN. MATERIALS AND METHODS: Betanin was isolated from fruits of Opuntia elatior Mill (Cactaceae) and structure was elucidated using spectroscopy (UV, IR, 1H-NMR and mass). STZ was injected intraperitoneally with single dose of 50mg/kg for diabetes induction. In order to develop DN the animals were left in diabetes condition without any treatment during the following 4 weeks. Betanin (25, 50 and 100mg/kg/day) and lisinopril (5mg/kg/day, reference compound) were orally administered for 8 weeks after the induction of DN. Renal function, blood glucose, serum creatinine, blood urea nitrogen (BUN) and antioxidant enzyme activities in the kidney tissue were measured. Kidney tissue samples were used for glomerulosclerosis, tubulointerstitial fibrosis and morphometric studies. The expression of transforming growth factor-beta (TGF-ß), type IV collagen, alpha-smooth muscle actin (α-SMA) and E-cadherin in kidney tissue were evaluated using reverse transcription-polymerase chain reaction, and immunohistochemistry. RESULTS: Betanin was successfully isolated from fruits of Opuntia elatior Mill (Cactaceae) and purified by column chromatography. The results showed that betanin attenuated diabetic kidney injury by significantly inhibiting proteinuria, blood glucose, serum creatinine and BUN levels and restored antioxidant enzyme activities in kidney tissue. Histological studies exhibited that betanin treatment reduced the glomerular surface area, glomerulosclerosis and tubulointerstitial fibrosis. Furthermore, betanin modulated mRNA and protein expression of TGF-ß, type IV collagen, α-SMA and E-cadherin in kidney. CONCLUSIONS: The results conclude that betanin can effectively suppress renal fibrosis in DN, and may slow down the progression to end-stage renal disease by regulating TGF-ß signal pathway.

Comparative Study of Betacyanin Profile and Antimicrobial Activity of Red Pitahaya (Hylocereus polyrhizus) and Red Spinach (Amaranthus dubius).

Betacyanins are reddish to violet pigments that can be found in red pitahaya (Hylocereus polyrhizus) and red spinach (Amaranthus dubius). This study investigated the impact of sub-fractionation (solvent partitioning) on betacyanin content in both plants. Characterization of betacyanins and evaluation of their antimicrobial activities were also carried out. Betanin was found in both plants. In addition, isobetanin, phyllocactin and hylocerenin were found in red pitahaya whereas amaranthine and decarboxy-amaranthine were found in red spinach. Sub-fractionated red pitahaya and red spinach had 23.5 and 121.5 % more betacyanin content, respectively, than those without sub-fractionation. Sub-fractionation increased the betanin and decarboxy-amaranthine content in red pitahaya and red spinach, respectively. The betacyanin fraction from red spinach (minimum inhibitory concentration [MIC] values: 0.78-3.13 mg/mL) demonstrated a better antimicrobial activity profile than that of red pitahaya (MIC values: 3.13-6.25 mg/mL) against nine Gram-positive bacterial strains. Similarly, the red spinach fraction (MIC values: 1.56-3.13 mg/mL) was more active than the red pitahaya fraction (MIC values: 3.13-6.25 mg/mL) against five Gram-negative bacterial strains. This could be because of a higher amount of betacyanin, particularly amaranthine in the red spinach.

Betacyanins enhance vitexin-2-O-xyloside mediated inhibition of proliferation of T24 bladder cancer cells.

Betacyanins (BC) were purified from beetroot (Beta vulgaris var. rubra L.) and tested, alone or in combination with vitexin-2-O-xyloside (XVX) from Beta vulgaris var. cicla L., for their ability to reduce the proliferation rate in T24 bladder cancer cells. Combination of BC and XVX exhibited a synergistic effect concerning the inhibition of proliferation in T24 cancer cells at 24 and 48 h but not after 72 h of incubation. The induction of apoptosis was evidenced by means of fluorescence activated cell sorting (FACS) analysis, as well as through the increase in caspase 3 and 8 activities. Using RTqPCR experiments, it was shown that the combination of XVX + BC was able to enhance the expression levels of pro-apoptotic BAX and downregulate anti-apoptotic BIRC5 (survivin), as well as pro-survival CTNNB1 (ß-catenin). The most evident effect of BC was the increase of the activity of caspase 8, leading to induction of extrinsic apoptosis. Moreover, XVX, BC and their combination showed no cytotoxic effect on normal human skin NCTC 2544 keratinocytes. These results demonstrated the efficacy and the mechanisms of the action of BC and XVX, extracted from edible plants, and suggested that a diet or a nutrition supplement, enriched with these bioactive molecules, could be used in the prevention of human bladder cancer.