Biosynthesis of Silica Nanoparticles Using the Leaf Extract of Punica granatum and Assessment of Its Antibacterial Activities Against Human Pathogens.

Plant-mediated nanoparticle synthesis is a revolutionary technique with numerous applications in fields, such as agriculture, food processing, and medicine. This study reports that Punica granatum leaf extract is capable of the green and eco-friendly synthesis of silica nanoparticles that provides a simple, cost-effective, and efficient methodology. P. granatum leaf extract was employed as a capping and stabilizing agent for the formation of silica nanoparticles, which were synthesized by a biological method using tetra ethyl ortho silicate. Biosynthesized silica nanoparticles are characterized by X-ray diffraction analysis, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and Fourier transform infrared spectroscopy. All the analyses and characterization determined that the particles were spherical in shape and amorphous in nature with an average size of 12 nm. P. granatum-assisted silica nanoparticles were tested for antibacterial activity by a well-diffusion method against two-gram negative bacterial pathogens (E. coli and Salmonella sp.). The antibacterial studies prove that P. granatum-assisted silica nanoparticles have good antibacterial properties. These studies will help us find a new nano-drug or medicine against multidrug-resistant bacteria.

Amelioration of diabetic nephropathy using pomegranate peel extract-stabilized gold nanoparticles: assessment of NF-κB and Nrf2 signaling system.

BACKGROUND: Diabetic nephropathy (DN), an end-stage renal disorder, has posed a menace to humankind globally, because of its complex nature and poorly understandable intricate mechanism. In recent times, functional foods as potential health benefits have been gaining attention of consumers and researchers alike. Rich in antioxidants, the peel and seed of pomegranate have previously demonstrated protection against oxidative-stress-related diseases, including cardiovascular disorders, diabetes, and cancer. PURPOSE: This study was designed to investigate the ameliorative role of pomegranate peel extract-stabilized gold nanoparticle (PPE-AuNP) on streptozotocin (STZ)-induced DN in an experimental murine model. METHODS: Following the reduction methods, AuNP was prepared using the pomegranate peel ellagitannins and characterized by particle size, physical appearance, and morphological architecture. Modulatory potential of PPE-AuNP was examined through the plethora of biochemical and high throughput techniques, flow cytometry, immunoblotting, and immunofluorescence. RESULTS: The animals treated with PPE-AuNP markedly reduced the fasting blood glucose, renal toxicity indices, and serum TC and TG in a hyperglycemic condition. As evident from an increased level of plasma insulin level, PPE-AuNP normalized the STZ-induced pancreatic ß-cell dysfunction. The STZ-mediated suppression of endogenous antioxidant response was restored by the PPE-AuNP treatment, which reduced the generation of LPO as well as iROS. Furthermore, the hyperglycemia-mediated augmentation of protein glycation, followed by the NOX4/p-47phox activation, diminished with the application of PPE-AuNP. The histological and immunohistochemical findings showed the protective efficacy of PPE-AuNP in reducing STZ-induced glomerular sclerosis and renal fibrosis. In addition, it reduced proinflammatory burden through the modulation of the MAPK/NF-κB/STAT3/cytokine axis. Simultaneously, PI3K/AKT-guided Nrf2 activation was evident upon the PPE-AuNP application, which enhanced the antioxidant response and maintained hyperglycemic homeostasis. CONCLUSION: The findings indicate that the use of PPE-AuNPs might act as an economic therapeutic remedy for alleviating DN.

Quantification of punicalagins in commercial preparations and pomegranate cultivars, by liquid chromatography-mass spectrometry.

BACKGROUND: Pomegranate (Punica granatum L.) - a delicious fruit once used in Ayurvedic medicine - is now largely known for the antioxidant properties of its juice, which has also been considered to have health benefits against diseases such as cancer and cardiovascular diseases. These beneficial effects are associated with the fruit's high content of polyphenolic compounds. High demand and lower production levels drive pomegranate prices up, which leads to the possibility of pomegranate products being adulterated, diluted or substituted. To ensure the presence of pomegranate in various preparations labeled as containing pomegranate, a simple method was developed to screen and quantify the specific punicalagins by mass spectrometry. RESULTS: The present method was used to analyze several pure and mixed beverages from the US market, and also to quantify punicalagins in the juice of 14 pomegranate cultivars. Punicalagins were detected in all cultivars, with higher concentrations in whole fruit juices compared with aril juices. Amongst the 20 commercial beverages, punicalagins were not detected in four preparations. CONCLUSION: The liquid chromatographic-mass spectrometric method presented herein enables an easy and rapid quantification of the specific punicalagins. The latter was detected in all cultivar samples, thus supporting that punicalagin is a suitable marker of these 14 pomegranate cultivars in commercial juices. Absence of the specific marker in four commercial preparations shows the necessity of having simple and rapid methods to evaluate the presence of pomegranate in preparations. © 2019 Society of Chemical Industry.

Benchmarking laboratory-scale pomegranate vinegar against commercial wine vinegars: antioxidant activity and chemical composition.

BACKGROUND: There is growing interest in the beneficial health effects of certain fruits, such as pomegranate, and their by-products, like vinegar. Vinegars contain antioxidant compounds such as polyphenols, which can scavenge free radicals in the body. In this study, the antioxidant properties (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid and 2,2-diphenyl-1-picrylhydrazyl scavenging capacities) and global polyphenolic composition of a new functional product, namely a pomegranate vinegar produced in the laboratory from the Gabsi variety, were evaluated and compared with those of commercial wine vinegars (Sherry and Rioja red wine Spanish vinegars). The evolution of the polyphenolic and volatile compositions during production of the pomegranate vinegar was also studied. RESULTS: The results indicate that pomegranate contained a relatively stable total phenolic content that is suitable for the elaboration of vinegar and that this product had an antioxidant capacity comparable to, or even better than, that observed for aged wine vinegars. Regarding the volatiles profile, a high proportion of esters (close to 50%), in relation to alcohols and fatty acids, was observed for pomegranate vinegar, and this is consistent with the high red fruits character identified by sensory analysis. CONCLUSION: Pomegranate vinegar produced in the laboratory has an antioxidant activity and a global phenolic content similar to those of red wine vinegars and higher than most of the white wine vinegars studied. The pomegranate vinegar retained the red fruit sensory character of pomegranate fruit. © 2018 Society of Chemical Industry.

A Comparative Study Between Labeling and Reality: The Case of Phytochemical Composition of Commercial Pomegranate-Based Products.

Manufacturers are deeply involved in the development of new pomegranate-based products, which have acquired great prestige due to many studies proving their potential health benefits. Commercial pomegranate products including capsules and supplements and juices and nectars were assayed. The contents of the key phytochemical compounds (punicalagin, ellagic acid, and total polyphenolic content) and the associated antioxidant capacity (DPPH• , ABTS•+ , and FRAP) were analyzed. The experimental ranges of punicalagin and ellagic acid contents were 0.96 to 308 mg/g and 0.09 to 13.1 mg/g, respectively. Punicalagin content was positively correlated (P < 0.001) with DPPH• and total polyphenolic content values. No significant (P > 0.05) correlation was observed among ellagic acid with the antioxidant capacity. The labeling standardization of these products is necessary due to the wide variability among "theoretically" similar pomegranate-based products.

Antimicrobial activity of pomegranate peel extracts as affected by cultivar.

BACKGROUND: Some studies have reported that different parts of the pomegranate fruit, especially the peel, may act as potential antimicrobial agents and thus might be proposed as a safe natural alternative to synthetic antimicrobial agents. The high tannin content, especially punicalagin, found in pomegranate extracts, has been reported as the main compound responsible for such antimicrobial activity. Because the pomegranate peel chemical composition may vary with the type of cultivar (sweet, sour-sweet and sour), pomegranates may also differ with respect to their antimicrobial capacity. RESULTS: The extract from PTO8 pomegranate cultivar peel had the highest antimicrobial activity, as well as the highest punicalagins (α and ß) and ellagic acid concentrations. In the results obtained from both antibacterial and antifungal activity studies, the sour-sweet pomegranate cultivar PTO8 showed the best antimicrobial activity, and the highest ellagic acid concentrations. CONCLUSION: The results of the present study suggest that ellagic acid content has a significant influence on the antimicrobial activity of the pomegranate extracts investigated. The pomegranate peel of the PTO8 cultivar is a good source of antifungal and antibacterial compounds, and may represent an alternative to antimicrobial agents of synthetic origin. © 2016 Society of Chemical Industry.

Effect of Pomegranate Hull Extract on Liver Neoplastic Changes in Rats: More than an Antioxidant.

Hepatocellular carcinoma (HCC) is the fifth most common cancer and the third leading cause of cancer-related mortality worldwide. The current work was designed to elucidate the molecular mechanisms underlying the antitumorigenic effect of pomegranate hull extract (PHE) in livers of rats exposed to the hepatocarcinogen diethyl nitrosamine (DENA) with emphasis on oxidative stress, proliferation, and apoptosis. Male albino rats were divided into three groups: normal control, DENA group, and PHE group. PHE was given to rats orally 3 times weekly for 10 wk, 4 wk before and 6 wk after DENA (200 mg/kg, single i.p. dose). The results indicated a prophylactic effect of PHE against neoplastic changes in the liver, which was evidenced by the decrease of tumor size, liver index, and the anti-apoptotic protein Bcl-2; and the increase of glutathione. PHE group also showed decreased expression of liver cyclin D1 and ß-catenin genes compared with DENA group. It is proved that PHE has antitumorigenic effect and could be a candidate for anticancer drugs.

Cost-effective imprinting combining macromolecular crowding and a dummy template for the fast purification of punicalagin from pomegranate husk extract.

The combination of molecular crowding and virtual imprinting was employed to develop a cost-effective method to prepare molecularly imprinted polymers. By using linear polymer polystyrene as a macromolecular crowding agent, an imprinted polymer recognizable to punicalagin had been successfully synthesized with punicalin as the dummy template. The resulting punicalin-imprinted polymer presented a remarkable selectivity to punicalagin with an imprinting factor of 3.17 even at extremely low consumption of the template (template/monomer ratio of 1:782). In contrast, the imprinted polymer synthesized without crowding agent, did not show any imprinting effect at so low template amount. The imprinted polymers made by combination of molecular crowding and virtual imprinting can be utilized for the fast separation of punicalagin from pomegranate husk extract after optimizing the protocol of solid-phase extraction with the recovery of 85.3 ± 1.2%.

Authentication of Punica granatum L.: Development of SCAR markers for the detection of 10 fruits potentially used in economically motivated adulteration.

The large commercial success of pomegranate increase the likelihood of economically motivated adulteration (EMA), which has been gradually spotted with the undeclared addition of anthocyanin-rich plants or cheaper fruit juices used as bulking and diluting agents. A method based on Sequence-Characterized Amplified Regions (SCARs) was developed to detect the presence of Aristotelia chilensis, Aronia melanocarpa, Dioscorea alata, Euterpe oleracea, Malus×domestica, Morus nigra, Sambucus nigra, Vaccinium macrocarpon, Vaccinium myrtillus, Vitis vinifera as bulking agents in Punica granatum. The method enabled the unequivocal detection of up to 1% of each adulterant, allowing the preemptive rejection of suspect samples. The recourse to such method may reduce the number of samples to be subjected to further phytochemical analyses when multiple batches have to be evaluated in a short time. Vice versa, it allows the cross-check of suspect batches previously tested only for their anthocyanin profile. The dimension of the amplicons is suitable for the analysis of degraded DNA obtained from stored and processed commercial material. Proper SCAR markers may represent a fast, sensitive, reliable and low-cost screening method for the authentication of processed commercial pomegranate material.

Bioactive properties of commercialised pomegranate (Punica granatum) juice: antioxidant, antiproliferative and enzyme inhibiting activities.

Pomegranate juice and related products have long been used either in traditional medicine or as nutritional supplements claiming beneficial effects. Although there are several studies on this food plant, only a few studies have been performed with pomegranate juice or marketed products. The aim of this work is to evaluate the antioxidant effects of pomegranate juice on cellular models using hydrogen peroxide as an oxidizing agent or DPPH and superoxide radicals in cell free systems. The antiproliferative effects of the juice were measured on HeLa and PC-3 cells by the MTT assay and pharmacologically relevant enzymes (cyclooxygenases, xanthine oxidase, acetylcholinesterase and monoamine oxidase A) were selected for enzymatic inhibition assays. Pomegranate juice showed significant protective effects against hydrogen peroxide induced toxicity in the Artemia salina and HepG2 models; these effects may be attributed to radical scavenging properties of pomegranate as the juice was able to reduce DPPH and superoxide radicals. Moderate antiproliferative activities in HeLa and PC-3 cancer cells were observed. However, pomegranate juice was also able to inhibit COX-2 and MAO-A enzymes. This study reveals some mechanisms by which pomegranate juice may have interesting and beneficial effects in human health.

Effects of ellagic acid-rich extract of pomegranates peel on regulation of cholesterol metabolism and its molecular mechanism in hamsters.

The study investigated the effect of pomegranates ellagic acid (PEA) on blood cholesterol and investigated its effects on LXR/RXR/PPAR-ABCA1 nuclear receptors-signaling pathways of cholesterol metabolism on molecular level in hamsters. In this experiment, hamsters were randomly divided into two groups: the first group (NG, n = 9) was always fed the normal diet, whereas the other group (HFG, n = 45) was fed a high fat diet during the first 4 weeks and then fed the normal diet for the last 4 weeks. In HFG, which was divided into five groups (n = 9) during the last 4 weeks, three groups were treated with PEA at 44 mg per kg bw, 88 mg per kg bw and 177 mg per kg bw, one group was treated with simvastatin at 1.77 mg per kg bw, and one was given sterile double-distilled water. The data validated that PEA dose-dependently decreased plasma total cholesterol and triglyceride level accompanied by a greater excretion of fecal bile acid. The result of RT-PCR revealed that PEA up-regulated liver X receptor (LXRα), peroxisome proliferator-activated receptor α (PPARα), peroxisome proliferator-activated receptor γ (PPARγ) and their downstream gene ATP-binding cassette transporter A1 (ABCA1), with no effect on retinoid X receptor (RXRα). PEA promoted cholesterol removal by enhancing fecal bile acid and up-regulation of the two pathways, LXR/PPAR-ABCA1. Moreover, PEA was stronger than simvastatin in some aspects.

Ethnopharmacological assessment of medicinal plants used against livestock infections by the people living around Indus River.

The present study was aimed to document detailed ethnopharmacological knowledge of medicinal plants against livestock infections of an unexplored remote region of Pakistan. Semistructured questionnaires were used for data collection. Total 43 plants belonging to 26 families were found to be used in ethnoveterinary practices. Seeds (29%) were found to be the most frequent plant part used followed by leaves (22%). Ethnoveterinary recipes were mostly prepared in the form of decoction and powdering. Informant consensus factor (Fic) results revealed high consensus for gastrointestinal (0.81), mastitis (0.82), and dermatological infections (0.80). Curcuma longa ranked first with highest fidelity level (FL) value (66%) followed by Trachyspermum ammi that ranked second (58%). Preference ranking (PR) results showed that Zingiber officinale, Punica granatum, Triticum aestivum, Gossypium hirsutum, and Withania coagulans were the most preferred species for the treatment of diarrhea. Direct matrix ranking (DMR) results showed that Morus alba, Melia azedarach, Withania coagulans, Cassia fistula, Azadirachta indica, and Tamarix aphylla were the multipurpose species of the region. We invite the attention of pharmacologists and chemists for further exploration of plants having high Fic, FL, and PR values in the present study. Conservation strategies should be adopted for the protection of multipurpose plant species.

Thermal stability of liquid antioxidative extracts from pomegranate peel.

BACKGROUND: Liquid extracts from pomegranate peel have the potential for use as natural antioxidant products. This study investigates the quality changes of liquid extracts before and after thermal treatment during sterilization and storage. Liquid pomegranate peel extracts were prepared, sterilized under ultra-high temperature (UHT) at 121 °C for 10 s and then stored at three temperatures (4, 25 and 37 °C) for up to 180 days. The industrial, color, UV-visible spectrum profile and antioxidant (phenolics) characteristics were measured. RESULTS: Thermal sterilization treatment had no negative effects on the industrial, color, spectral and antioxidant characteristics of the extracts. After 180 days, the extracts stored at 4 °C retained 67% of the initial total soluble phenolic content and 58% of the original scavenging activity. The major antioxidant components in the extracts (stored at 4 °C for 180 days) were gallic acid, punicalagin A, punicalagin B and ellagic acid having concentrations of 19.3, 197.2, 221.1 and 92.4 mg L⁻¹, respectively. CONCLUSION: The results show that liquid pomegranate peel extracts had acceptable thermal stability after sterilization and storage. The recommended storage condition of this product was low temperature.

Pomegranate juice adulteration by addition of grape or peach juices.

BACKGROUND: Pomegranate juice has gained a high reputation for its health properties and consequently is now a highly demanded product. However, owing to the limited production and high price of fresh pomegranates, adulteration of pomegranate juice seems to be happening. Hence it is imperative to establish criteria for detecting adulteration. RESULTS: Addition of grape juice significantly increased the contents of Ca, Mg and Fe and especially tartaric acid and proline and simultaneously decreased the content of K. Addition of peach juice up to 10% (v/v) only resulted in a significant increase in sucrose content. Regarding the volatile composition, adulteration of pomegranate juice with grape juice resulted in significant increases in acetic acid, isoamyl butyrate and especially 1-hexanol and linalool, while adulteration with peach juice resulted in significant increases in butyl acetate, isobutyl butyrate, benzyl acetate and especially isoamyl butyrate. CONCLUSION: The control protocols used in this study can serve as a basis for identification of pomegranate juice adulteration. It is important to highlight that it is necessary to simultaneously analyze and have results from several parameters to conclude that a particular pomegranate juice has been adulterated by mixing with another fruit juice.

Assessment of pomegranate wine lees as a valuable source for the recovery of (poly)phenolic compounds.

Wine lees are the residue formed at the bottom of recipients containing wine after fermentation and are mainly composed of yeast. They can be applied for the recovery of value-added phytochemicals owing to the ability of yeast to form molecular interactions with (poly)phenolic compounds. This study aimed to evaluate the potential use of lees obtained as by-products after winemaking of pomegranate (Punica granatum L.) juice. Pomegranate wine lees showed high both phenolic content (about 30 mg GAE/g dry matter) and antioxidant capacity by DPPH and ABTS(+) assays. The phytochemical screening of this by-product by UHPLC-ESI-MS(n) allowed the identification of up to 39 chemicals, being hydrolysable tannins and anthocyanins the predominant structures. Ellagic acid and gallic acid were present in high amounts. In addition, mineral composition was also assessed. Overall, pomegranate wine lees resulted in a promising source for the recovery of bioactive polyphenols with potential applications in different industrial fields.

Assessment of antioxidant, anti-inflammatory, anti-cholinesterase and cytotoxic activities of pomegranate (Punica granatum) leaves.

This study evaluated antioxidant, anti-inflammatory, anti-cholinesterase and cytotoxic activities of extracts with different polarities (hexane, dichloromethane, ethyl acetate, ethanol and methanol) obtained from Punica granatum leaves. Total phenolics (8.8-127.3mg gallic acid equivalent/g dry weight), flavonoids (1.2-76.9mg quercetin equivalent/g dry weight), tannins (63.7-260.8mg catechin equivalent/kg dry weight) and anthocyanins (0.41-3.73mg cyanidin-3-glucoside equivalent/g dry weight) of different extracts were evaluated. The methanolic extract presented a good IC50 by DPPH and ABTS assays (5.62 and 1.31mg/l respectively). The strongest 5-lipoxygenase (5-LOX), acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibition activities were obtained for the ethanol extract (IC50 values of 6.20, 14.83 and 2.65mg/l, respectively) and the best cytotoxic activity against MCF-7 cells was obtained for the methanol extract (IC50=31mg/l). These important biological activities showed that P. granatum leaves could be a potential source of the active molecules intended for applications in pharmaceutical industry, but only after additional in vivo experiments.

Preparation of low cost activated carbon from Myrtus communis and pomegranate and their efficient application for removal of Congo red from aqueous solution.

In this research, the potential applicability of activated carbon prepared from Myrtus communis (AC-MC) and pomegranate (AC-PG) as useful adsorbents for the removal of Congo red (CR) from aqueous solutions in batch method was investigated. The effects of pH, contact time, agitation time and amount of adsorbents on removal percentage of Congo red on both adsorbents were examined. Increase in pH up to 6 for AC-MC and pH 7 for AC-PG increase the adsorption percentage (capacity) and reach equilibrium within 30 min of contact time. Fitting the experimental data to conventional isotherm models like Freundlich, Langmuir, Tempkin and Dubinin-Radushkevich show that the experimental data fitted very well to the Freundlich isotherm for AC-MC and Langmuir isotherm for AC-PG. Fitting the experimental data to different kinetic models such as pseudo first-order, pseudo second-order, Elovich and intraparticle diffusion mechanism showed the applicability of a pseudo second-order with involvement of intraparticle diffusion model for interpretation of experimental data for both adsorbents. The adsorption capacity of AC-PG and AC-MC for the removal of CR was found to be 19.231 and 10 mg g(-1). These results clearly indicate the efficiency of adsorbents as a low cost adsorbent for treatment of wastewater containing CR.

Preparation method and stability of ellagic acid-rich pomegranate fruit peel extract.

A simple one-step purification using liquid-liquid extraction for preparing pomegranate peel extract rich in ellagic acid has been demonstrated. The method involved partitioning of the 10% v/v water in methanol extract of pomegranate peel between ethyl acetate and 2% aqueous acetic acid. This method was capable of increasing the ellagic acid content of the extract from 7.06% to 13.63% w/w. Moreover, the antioxidant activity of the extract evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay was also increased (ED(50) from 38.21 to 14.91 micro/mL). Stability evaluations of the ellagic acid-rich pomegranate peel extract in several conditions through a period of four months found that the extracts were stable either kept under light or protected from light. The extracts were also stable under 4 degrees +/- 2 degrees C, 30 degrees +/- 2 degrees C and accelerated conditions at 45 degrees C with 75% relative humidity. However, study on the effect of pH on stability of the extract in the form of solution revealed that the extract was not stable in all tested pH (5.5, 7 and 8). These results indicated that the ellagic acid-rich pomegranate peel extract was stable when it was kept as dried powder, but it was not stable in any aqueous solution.

Assessment of mutagenic and antimutagenic effects of Punica granatum in mice / Avaliação dos efeitos mutagênicos e antimutagênica de Punica granatum em camundongos

In the present study, the ability of Punica granatum ethanolic leaf extract (PGL) and Punica granatum ethanolic fruit extract (PGF) to induce mutagenicity or to modulate the genotoxic effects induced by the alkylating agent cyclophosphamide (CP) was evaluated. Swiss male mice were treated by gavage for 10 days with PGL or PGF (12.5, 25, 50, and 75 mg/kg/day) prior to exposure to CP (i.p. 200 mg/kg), 24 h after the end of the treatment. Initial observations revealed that normal mice treated with both extracts (12.5, 25, 50, and 75 mg/kg/day) showed a similar micronucleated polychromatic erythrocyte (MNPCE) frequency to that of the control group. Investigation of the protective effect of PGL and PGF based on data analysis revealed that, irrespective of dose or extract, oral administration of PGL or PGF for 10 days prior to exposure had reduced, in a dose-dependent manner, the frequency of MNPCE induced by CP in all groups studied. Higher reductions were observed at PGF doses of 50 and 75 mg/kg. Taken together, these results demonstrate that mice treated with P. granatum showed an absence of mutagenic effects and dose-dependent protective effects against CP-induced oxidative DNA damage.

No presente estudo investigamos o potencial do extrato etanólico das folhas da Punica granatum (PGFO) e do extrato etanólico dos frutos da Punica granatum (PGFR) de induzir mutagenicidade ou de proteger contra efeitos genotóxicos induzidos pela ciclofosfamida (CF). Camundongos machos Swiss foram tratados por 10 dias, via oral, com PGFO ou PGFR (12,5, 25, 50 e 75 mg/kg/dia), previamente a exposição à CF (i.p. 200 mg/kg) 24 horas após término do tratamento. Observamos que os animais tratados por 10 dias com ambos os extratos (12,5, 25, 50 e 75 mg/kg/dia) demonstraram a frequência de micronúcleo policromático eritrocitário (MNPCE) similar ao grupo controle. Quando aos efeitos protetores dos extratos foram investigados, a análise dos dados revelou que, independentemente da dose ou do extrato usado, a administração oral por 10 dias, previamente à exposição, reduziu, de forma dose-dependente, a frequência de MNPCE induzidos pela CF, em todos os grupos estudados. As maiores reduções foram observadas com PGFR nas doses de 50 e 75 mg/kg. Em conjunto, sob as condições testadas, camundongos tratados com P. granatum demonstraram ausência de efeitos mutagênicos e, de forma dose-dependente, efeitos protetores contra os danos oxidativos do DNA induzidos pela CF.

Transport behavior of ellagic acid of pomegranate leaf tannins and its correlation with total cholesterol alteration in HepG2 cells.

The aim of this study was to investigate whether ellagic acid in pomegranate leaf tannins could be transported into HepG2 cells and its transport behavior. High-performance liquid chromatography coupled with a 996 photodiode array detector at 254 nm was applied. The mobile phase was an acetonitrile-water solution (containing 0.1% triethylamine, pH 3.0; 16:64, v/v, for determining ellagic acid in cells). The flow rate was 0.8 mL/min. Cells were incubated with pomegranate leaf tannins with 100 and 50 microg/mL (containing 1.71 and 0.85 microg/mL of ellagic acid, respectively) for a specific time, then lysed and sonicated in methanol to extract intracellular ellagic acid. A 10 microL aliquot of sample was injected into the HPLC system to determine ellagic acid concentration. The results showed that ellagic acid in pomegranate leaf tannins could be transported into the cells, which was in correlation with total cholesterol alteration in the cells. This is the first time that the transport behavior of ellagic acid through HepG2 cells in vitro has been comprehensively demonstrated.