Effect of polyphenols extracts from Brassica vegetables on erythrocyte membranes (in vitro study)

The aim of this work was to estimate the in vitro effects of polyphenol extracts from Brassica vegetables (Brussels sprouts and red cabbage) on erythrocyte membranes with normal and high concentration of cholesterol. To determine the effect of phenolic compounds we prospectively studied cholesterol concentration, lipid peroxidation, membrane fluidity and ATPase activity. Polyphenol extracts from Brassica vegetables resulted in statistically significant reductions in cholesterol concentrations in hypercholesterolemic erythrocytes. For control erythrocytes, no significant reduction of cholesterol levels was observed for both extracts. Decreases in lipid peroxidation intensity were observed after incubation of hypercholesterolemic erythrocytes with the extracts. No changes in membrane fluidity for both extracts were noted for normal and hypercholesterolemic erythrocytes. The activity of ATPase decreased after incubation of normal and hypercholesterolemic erythrocytes with extract from Brassica vegetables. Our results indicate that polyphenols from red cabbage and Brussels sprout may directly influence erythrocyte membrane properties.     

Evaluation of the effect of Uncaria tomentosa extracts on the size and shape of human erythrocytes (in vitro)

In this study, we continued our investigations concerning the interaction of Uncaria tomentosa extracts with the human erythrocytes. The analysis of the size and shape of the erythrocytes by means of flow cytometry and phase contrast microscopy was performed. We executed our experiments using ethanolic and aqueous extracts from the leaves and bark of U. tomentosa. Disturbances were observed in the size and shape of the erythrocytes incubated with ethanolic and aqueous extracts at the concentrations of 100 μg/mL and 250 μg/mL, respectively. The observed changes were probably related to the entry of polyphenolic compounds contained in U. tomentosa extracts into erythrocyte membrane. Externalization of phosphatidylserine on the erythrocytic surfaces was also noticed during incubation with extracts at concentration of 250 μg/mL. We concluded that all of the extracts examined induced changes in the erythrocyte membrane properties, whereas ethanolic extracts from bark induced the most significant changes. The possible binding of polyphenols to the erythrocyte surface may have accounted for the protective properties of extracts against haemolysis of RBCs, which was observed in our previous study (Bors et al., 2011), but considerable incorporation of polyphenols into cell membranes can result in disturbance of phosphatidylserine transport and changes in erythrocyte shape. Nevertheless the results of the investigations showed that considerable morphological changes appear only as a result of erythrocyte exposure to high concentrations (50 ppm and 100 ppm) of the extracts studied, thus they should not lead to clinical erythrocytic damage if recommended doses of U. tomentosa preparations are administrated.     

Surface and emulsifying properties of raw ethanolic extracts from legumes containing soyasaponins and proteins

The development of new amphiphilic compounds of natural origin is a challenging field of research in surfactant science. In this scenario, surfactants obtained from renewable organic sources is a valuable option to be exploited. The present work proposes raw ethanolic extracts containing soyasaponins, proteins and polysaccharides from legumes as surface active mixtures and stabilizers for emulsified systems. These extracts are produced by a green extraction process using an hydroalcoholic mixture (70:30 w/w) as solvent and six different legumes soybean, lentils, peas, beans, chickpeas, wild peas as starting materials. After freeze-drying, extracts were characterized in terms of total soyasoponin (from 1377 to 7354 mg/100 g), soyasaponin I (from 15 to 1459 mg/kg), soyasaponin VI (from 13 to 1076 mg/kg) and protein (16–28 g/100 g) contents. All extracts aqueous solutions show good surface active properties, being able to reduce the air-water surface tension to values comparable to those of the commonly employed surfactants (28–33 mN/m). Differences are observed in terms of emulsifying ability, since only lentils and soybeans extracts, thanks to their high soyasaponins and protein content, can act as stabilizers for emulsions containing up to 10% w/w of ethyl oleate (as oil phase) over six months of storage at 4 °C. These extracts are promising amphiphilic natural mixtures, potentially employable for food or pharmaceutical applications. Their relevance is also related to the possibility of using legumes derived from industrial wastes of food manufacturing, thereby giving valorization of by-products and residues in the context of a “circular economy” model.     

Antioxidant and antitumor activities of Artemisia campestris and Thymelaea hirsuta from southern Tunisia

The essential oil of Artemisia campestris and the ethanol-water, hexane and water extracts of A. campestris and Thymelaea hirsuta collected in southern of Tunisia were investigated for their antioxidant (DPPH, ABTS and beta-carotene methods) and antitumor growth inhibition of human colon cancer HT-29 cells using MTT test activities.All the A. campestris extracts tested at high concentrations (100 μg/ml) showed activity ranging from 19.5% for essential oil to 64.4% of negative control growth for infusion extract, except the hexane extract. With T. hirsuta, all the extracts tested (hexane and ethanol-water), except the infusion extract, also exhibited antitumor activity (58.2% and 65.5% of control growth respectively).The ethanol-water and infusion extracts of A. campestris showed higher antioxidant activity, polyphenol and flavonoid contents than those of T. hirsuta. These results show that there is a positive correlation between the antitumor activity and the antioxidant activity, and of these two activities and with the levels of polyphenols and flavonoids.The essential oil and the other extracts of A. campestris, which exhibited significant antitumor activity against the HT-29 cells deserve further research into the chemoprevention and treatment of colon cancer.     

Characterization of nine polyphenols in fruits of Malus pumila Mill by high-performance liquid chromatography

Polyphenols are important bioactive substances in apple. To explore the profiles of the nine representative polyphenols in this fruit, a high-performance liquid chromatography method has been established and validated. The validated method was successfully applied for the simultaneous characterization and quantification of these nine apple polyphenols in 11 apple extracts, which were obtained from six cultivars from Shaanxi Province, China. The results showed that only abscission of the Fuji apple sample was rich in the nine apple polyphenols, and the polyphenol contents of other samples varied. Although all the samples were collected in the same region, the contents of nine polyphenols were different. The proposed method could serve as a prerequisite for quality control of Malus products.     

Dapsone hydroxylamine induces premature removal of human erythrocytes by membrane reorganization and antibody binding

BACKGROUND AND PURPOSE

N-hydroxylation of dapsone leads to the formation of the toxic hydroxylamines responsible for the clinical methaemoglobinaemia associated with dapsone therapy. Dapsone has been associated with decreased lifespan of erythrocytes, with consequences such as anaemia and morbidity in patients treated with dapsone for malaria. Here, we investigated how dapsone and/or its hydroxylamine derivative (DDS-NHOH) induced erythrocyte membrane alterations that could lead to premature cell removal.

EXPERIMENTAL APPROACH

Erythrocytes from healthy donors were subjected to incubation with dapsone and DDS-NHOH for varying times and the band 3 protein tyrosine-phosphorylation process, band 3 aggregation, membrane alteration and IgG binding were all examined and compared with erythrocytes from two patients receiving dapsone therapy.

KEY RESULTS

The hydroxylamine derivative, but not dapsone (the parent sulphone) altered membrane protein interactions, leading both to aggregation of band 3 protein and to circulating autologous antibody binding, shown in erythrocytes from patients receiving dapsone therapy. The band 3 tyrosine-phosphorylation process can be used as a diagnostic system to monitor membrane alterations both in vitro, assessing concentration and time-dependent effects of DDS-NHOH treatment, and in vivo, evaluating erythrocytes from dapsone-treated patients, in resting or oxidatively stimulated conditions.

CONCLUSIONS AND IMPLICATIONS

DDS-NHOH-induced alterations of human erythrocytes can be directly monitored in vitro by tyrosine-phosphorylation level and formation of band 3 protein aggregates. The latter, together with antibody-mediated labelling of erythrocytes, also observed after clinical use of dapsone, may lead to shortening of erythrocyte lifespan.     

Effects of soyasaponin I and soyasaponins-rich extract on the Alternariol-induced cytotoxicity on Caco-2 cells

Alternariol (AOH) is a mycotoxin produced by Alternaria spp. Soyasaponin I (Ss-I) is present naturally in legumes, and it has antioxidant properties. Cytotoxic and genotoxic effects of AOH have been demonstrated previously in vitro. In the present study, the cytotoxicity of AOH, Ss-I, and soyasaponins-rich extract from lentils was investigated; as well as, the cytoprotective effects of Ss-I and lentil extracts against AOH induced-cytotoxicity on Caco-2 cells. Cytotoxicity was carried out using MTT and PC assays (AOH: 3.125–100 µM, Ss-I: 3.125–50 µM, and lentil extracts: 1:0–1:32) during 24 h of exposure. Only AOH showed cytotoxic effect. The reduction in cell proliferation ranged from 25% to 47%. Simultaneous combination of Ss-I with AOH (1:1) increased cell proliferation (35%) compared to AOH tested alone. The Ss-I and extracts showed synergistic cytoprotective effects against cytotoxicity induced by AOH on Caco-2 cells. Food commodities containing Ss-I could contribute to diminish the toxicological risk that natural contaminant as AOH in diet can produce to humans.     

A comparative study of ofloxacin and ciprofloxacin erythrocyte distribution

The present work deals with the in vitro and in vivo distribution of ofloxacin and ciprofloxacin in erythrocytes. In vitro studies were carried out in standard solutions prepared using fresh blood for a concentration range between 100 and 0.25 μg mL⁻¹. A 5 mg kg⁻¹ bolus dose was administered to rabbits and erythrocyte and plasma kinetics were determined over 8 h. A linear model was used to establish the relationship between plasma and erythrocyte concentrations of both quinolones in vitro. The mean partition coefficient values obtained were 1.04±0.02 and 1.32±0.03 for ofloxacin and ciprofloxacin, respectively. A decrease in the ciprofloxacin partition coefficient was observed at higher concentrations. Values ranged between 2.54±0.40 and 1.38±0.15 as the concentrations increased. The partition coefficients obtained from the linear relationship between plasma and erythrocyte concentrations established from the in vivo data were 0.80±0.58 for ofloxacin and 0.61±0.30 for ciprofloxacin. In vivo plasma and erythrocyte data analysis was performed by a deconvolution method and the theoretical transfer curves in erythrocytes were estimated. The distribution of both quinolones to erythrocytes is very rapid, probably due to a high permeability of erythrocyte membranes to these drugs. This was also confirmed by the parallelism between plasma and erythrocyte kinetics. © 1998 John Wiley & Sons, Ltd.     

Human erythrocytes are affected in vitro by extracts of Ugni molinae leaves.

Ugni molinae Turcz, also known as "Murtilla", is a plant that grows in the south of Chile. Infusions of their leaves have long been used in traditional native herbal medicine. The chemical composition of the leaves indicates the presence of polyphenols, which have antioxidant properties. In order to evaluate the mechanisms of their antioxidant properties and the toxicity of the aqueous extracts of leaves, the extracts were induced to interact with human red cells, their isolated unsealed membranes (IUM) and large unilamellar vesicles (LUV) of dimyristoylphosphatidyltidylcholine (DMPC), representative of phospholipid classes located in the outer monolayer of the erythrocyte membrane. Scanning electron microscopy (SEM) observations indicated that the extracts achieved a significant alteration in the shape of the erythrocytes as they changed their discoid shape to echinocytes. According to the bilayer couple hypothesis, the shape change indicates that the polyphenols were located in the outer moiety of the red cell membrane. This conclusion was confirmed by the fluorescence experiments performed in IUM and DMPC LUV. In fact, the extracts produced slight initial increases followed by sharp decreases at higher concentrations in the anisotropy and general polarization parameters. These results imply that the extracts induced structural perturbations in the acyl chain and polar group packing arrangements of the erythrocyte IUM and DMPC LUV lipid bilayers: first ordering and afterwards disordering them as the extract concentration increased.     

Hemolytic properties of hexachlorophene and related chlorinated bisphenols

Although hexachlorophene (HCP) and other chlorinated bisphenols are not hemolytic in vivo, even when administered at lethal doses, incubation of these compounds with nucleated and non-nucleated erythrocytes in vitro produces hemolysis. Chlorinated bisphenols are effective hemolytic agents at concentrations of 2 × 10^?5 to 2 × 10^?4 M. The hemolytic effect of the chlorinated bisphenols is characteristic of the erythrocyte donor species rather than dependent on the presence or absence of nucleation in the red cells. Of all the species tested, fish erythrocytes are the most susceptible and turkey erythrocytes are the most resistant to hemolysis by HCP though both are nucleated. No correlation was found between the hemolytic activity of the chlorinated bisphenols and their lipophilic character or the degree of ionization (pKa). Whole plasma or solutions of human and bovine serum albumin, hemoglobin and other proteins protected the erythrocytes against hemolysis induced by chlorinated bisphenols. The protection varied somewhat with the chlorinated bisphenol in question and differed among proteins of the same type from different species.     

Protection against hydrogen peroxide induced oxidative damage in rat erythrocytes by Mangifera indica L. peel extract

Phytochemicals such as polyphenols and carotenoids are gaining importance because of their contribution to human health and their multiple biological effects such as antioxidant, antimutagenic, anticarcinogenic and cytoprotective activities and other therapeutic properties. Mango peel is a major by-product in pulp industry and it contains various bioactive compounds like polyphenols, carotenoids and others. In the present study, the protective effect of peel extracts of unripe and ripe mango fruits of two varieties namely, Raspuri and Badami on hydrogen peroxide induced hemolysis, lipid peroxidation, degradation of membrane proteins and its morphological changes are reported. The oxidative hemolysis of rat erythrocytes by hydrogen peroxide was inhibited by mango peel extract in a dose dependent manner. The IC₅₀ value for lipid peroxidation inhibition on erythrocyte ghost membrane was found to be in the range of 4.5–19.3 μg gallic acid equivalents. The mango peel extract showed protection against membrane protein degradation caused by hydrogen peroxide. Morphological changes to erythrocyte membrane caused by hydrogen peroxide were protected by mango peel extract. The results demonstrated that mango peel extracts protected erythrocytes against oxidative stress and may impart health benefits and it could be used as a valuable food ingredient or a nutraceutical product.     

Antihemolytic and antioxidant properties of pearl powder against 2,2’-azobis(2-amidinopropane) dihydrochloride-induced hemolysis and oxidative damage to erythrocyte membran elipids and proteins

Pearl powder, a well-known traditional mineral medicine, is reported to be used for well-being and to treat several diseases from centuries in Taiwan and China. We investigated the in vitro antihemolytic and antioxidant properties of pearl powder that could protect erythrocytes against 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative damage to membrane proteins/lipids. Human erythrocytes were incubated with different concentrations of pearl powder (50–200 μg/mL) for 30 minutes and then exposed to AAPH for 2–6 hours. We found that AAPH alone time dependently increased the oxidative hemolysis of erythrocytes, while pearl powder pretreatment substantially inhibited the hemolysis in a concentration-/time-dependent manner. AAPH-induced oxidative damage to erythrocyte membrane lipids was evidenced by the elevated malondialdehyde (MDA) levels. However, pearl powder remarkably inhibited the malondialdehyde formation, and the 200 μg/mL concentration showed almost similar malondialdehyde values to the control. Furthermore, pearl powder suppressed the AAPH-induced high-molecular-weight protein formation and concomitantly increased the low-molecular-weight proteins in erythrocytes. Antioxidant potential that was measured as superoxide dismutase activity and glutathione content was significantly dropped by AAPH incubation, which suggests the vulnerability of erythrocytes to AAPH-induced oxidative stress. Noteworthy, erythrocytes pretreated with pearl powder showed restored superoxide dismutase activity and glutathione levels against AAPH-induced loss. Our findings conclude that pearl powder attenuate free radical-induced hemolysis and oxidative damage to erythrocyte membrane lipids/proteins. The potent antioxidant property of pearl powder may offer protection from free radical-related diseases.     

Effect of selenium enrichment on antioxidant activities and chemical composition of Lentinula edodes (Berk.) Pegl. mycelial extracts

Preparations derived from Lentinula edodes (Berk.) Pegl. mycelium are worldwide used as dietary supplements containing compounds active as immune system enhancers, demonstrating chemopreventive and anticancer activity. L. edodes mycelium enriched with organic forms of selenium like selenized yeast possess putative, higher cancer preventive properties. The objective of this study was to test the effect of enrichment in selenium on antioxidant, reducing and free radical scavenging activity of water and alcohol extracts from mycelium of L. edodes (Berk.). To elucidate the cause of enhanced antioxidant activity of extracts, a preliminary selenium speciation by specific oxido-reduction reaction was performed. Se-enrichment enhanced antioxidant activity, reducing power and free radical scavenging effect of mycelial extracts by almost 100–400%. Increase of activity was particularly high for diluted extracts (concentrations 0.1–0.5 mg/ml). The chemical composition of extracts from both Se-enriched and non-enriched mycelium was compared by determination of polyphenols, proteins, carbohydrates and lipids. Results showed that Se-enrichment enhanced antioxidant activities of mycelial extracts, likely by high amounts of organic Se-compounds (–II oxidation state) and elemental red selenium, and by increased polyphenols content. Our results suggest that Se-enrichment is a good method for enhancement of important activities of human dietary supplements, including Shiitake preparations.     

Antihemolytic and antioxidant properties of pearl powder against 2,2′-azobis(2-amidinopropane) dihydrochloride-induced hemolysis and oxidative damage to erythrocyte membrane lipids and proteins

Pearl powder, a well-known traditional mineral medicine, is reported to be used for well-being and to treat several diseases from centuries in Taiwan and China. We investigated the in vitro antihemolytic and antioxidant properties of pearl powder that could protect erythrocytes against 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative damage to membrane proteins/lipids. Human erythrocytes were incubated with different concentrations of pearl powder (50–200 μg/mL) for 30 minutes and then exposed to AAPH for 2–6 hours. We found that AAPH alone time dependently increased the oxidative hemolysis of erythrocytes, while pearl powder pretreatment substantially inhibited the hemolysis in a concentration-/time-dependent manner. AAPH-induced oxidative damage to erythrocyte membrane lipids was evidenced by the elevated malondialdehyde (MDA) levels. However, pearl powder remarkably inhibited the malondialdehyde formation, and the 200 μg/mL concentration showed almost similar malondialdehyde values to the control. Furthermore, pearl powder suppressed the AAPH-induced high-molecular-weight protein formation and concomitantly increased the low-molecular-weight proteins in erythrocytes. Antioxidant potential that was measured as superoxide dismutase activity and glutathione content was significantly dropped by AAPH incubation, which suggests the vulnerability of erythrocytes to AAPH-induced oxidative stress. Noteworthy, erythrocytes pretreated with pearl powder showed restored superoxide dismutase activity and glutathione levels against AAPH-induced loss. Our findings conclude that pearl powder attenuate free radical-induced hemolysis and oxidative damage to erythrocyte membrane lipids/proteins. The potent antioxidant property of pearl powder may offer protection from free radical-related diseases.     

Tobramycin-induced modulation of macrophage Fc-receptors

Tobramycin is an aminoglycoside that is used therapeutically for chronic pulmonary infection that occurs in cystic fibrosis patients. The effect of this antibiotic on Fc-receptor expression was investigated using rat alveolar macrophages. When macrophage monolayers were incubated for 30 min with low concentrations of tobramycin (0.03-2.4 micrograms/ml) similar to those found in the cystic fibrosis lung, Fc-receptor expression was enhanced. This effect was dose-, temperature- and time-dependent. After 1 h pre-incubation antibody-labelled erythrocyte (EAG) binding returned to normal values and after 2 h inhibition was observed. When culture supernatants prepared from antibiotic-treated macrophages were incubated with EAG for 30 min, binding of these cells to the macrophages was decreased. This inhibition was directly proportional to the amount of supernatant added to the red cells and was greatest when the supernatant had been prepared from macrophages incubated with the antibiotic for 1 h. When macrophage monolayers were incubated for 1 h with supernatants from tobramycin-treated macrophages subsequent binding of EAG was increased. The supernatants also caused the agglutination of antibody-sensitised erythrocytes. This activity was lost after the supernatants had been absorbed against EAG. It is proposed that the supernatants contain released Fc-receptors and that tobramycin causes an increase in the shedding of Fc-receptors from the macrophage membrane. The mechanism of action of tobramycin and the in vivo relevance of these results is discussed.     

Zum Vorkommen von Hämolysinen in Pilzen der GattungAmanita

Phallolysin, the high-molecular weight hemolysin fromAmanita phalloides, was present in high activity in each of 42 samples ofA. phalloides. The toadstools were collected in 19 different locations during a period of 4 years. Aqueous extracts of 12 furtherAmanita species of differing origin were tested for hemolytic activity. Several samples of each species — altogether 99 — were tested on rabbit and human erythrocytes. Hemolysins were present inA. muscaria, A. gemmata, A. verna, A. citrina, A. porphyria, A. spissa, A. echinocephala and invariably and in high activity inA. rubescens. According to the present results and to unpublished observations, the hemolysin ofA. rubescens differed from Phallolysin ofA. phalloides. The extracts ofA. muscaria also produced agglutination of red blood cells.     

Extraction and quantification of polyphenols from kinnow (Citrus reticulate L.) peel using ultrasound and maceration techniques

An investigation was carried out to extract polyphenols from the peel of kinnow (Citrus reticulate L.) by maceration and ultrasound-assisted extraction (UAE) techniques. The antioxidant potential of these polyphenols was evaluated using ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and superoxide radical scavenging assays; and their antimicrobial activity was assessed against bacterial strains Staphyloccoccus aureus, Bacillus cereus, and Salmonella typhimurium. The highest extraction yield was obtained through the solvent ethanol at 80% concentration level, whereas UAE was a more efficient technique and yielded comparatively higher polyphenol contents than maceration. Maximum polyphenols were extracted with 80% methanol [32.48 mg gallic acid equivalent (GAE)/g extract] using UAE, whereas minimum phenolics (8.64 mg GAE/g extract) were obtained with 80% ethyl acetate through the maceration technique. Elevated antioxidant activity of kinnow peel extracts was exhibited in three antioxidant assays, where 80% methanolic extracts showed the highest antioxidant activity (27.67 ± 1.11mM/100 g for FRAP) and the highest scavenging activity, 72.83 ± 0.65% and 64.80 ± 0.91% for DPPH and superoxide anion radical assays, respectively. Strong correlations between total polyphenols and antioxidant activity were recorded. Eleven phenolic compounds—including five phenolic acids and six flavonoids—were identified and quantified by high performance liquid chromatography. Ferulic acid and hesperidin were the most abundant compounds whereas caffeic acid was the least abundant phenolic compound in kinnow peel extracts. Maximum inhibition zone was recorded against S. aureus (16.00 ± 0.58 mm) whereas minimum inhibition zone was noted against S. typhimurium (9.00 ± 1.16 mm). It was concluded that kinnow mandarin peels, being a potential source of phenolic compounds with antioxidant and antimicrobial properties, may be used as an ingredient for the preparation of functional foods.     

The action of six lipid perturbing substances on hormone receptor adenylate cyclase complex in turkey erythrocytes and intestinal mucosa cells

Adenylate cyclase and beta-receptor binding in turkey erythrocytes is studied under the influence of dioctyl sodium sulphosuccinate (DSS), sodium dodecyl sulphate (SDS), sodium deoxycholate (DOC), filipin, oxyphenisatine and ethanol. Intact and lysed erythrocytes and erythrocyte membranes were investigated. In general the fluoride stimulated enzyme was most resistant to the action of the substances followed by the ligand binding, the hormone stimulated enzyme being most sensitive to the action of the substances. However, in certain ranges of concentrations, DSS, SDS, filipin and oxyphenisatine could further stimulate the fluoride stimulated enzyme. The adenylate cyclase in intestinal mucosa cells was studied in the presence of DSS, SDS, DOC and oxyphenisatine. The fluoride stimulated enzyme was here less stable than the basal enzyme activity, and two of the substances (SDS and oxyphenisatine) stimulating the erythrocyte enzyme could not stimulate the fluoride stimulated intestinal mucosa cell enzyme.     

Activation of the erythrocyte plasma membrane redox system by resveratrol: a possible mechanism for antioxidant properties

Resveratrol is one of the most widely studied of all the plant-produced polyphenols and has diverse, beneficial health effects including anti-cancer and cardio-protective effects. Many of the biological actions of this polyphenol have been attributed to its antioxidant properties. Erythrocytes contain a plasma membrane redox system (PMRS), which transfers electrons from intracellular donors (NADH and/or ascorbate (ASC)) to extracellular acceptors. There is evidence that the intracellular ASC donates electrons to extracellular ascorbate free radicals (AFRs) via the PMRS, which encompasses an AFR reductase; such a redox system enables the cells to effectively counteract oxidative processes.We present evidence to show that human erythrocytes take up resveratrol, and once inside the cell, resveratrol can donate electrons to extracellular electron acceptors through the erythrocyte PMRS and AFR reductase. Incubating human erythrocytes with resveratrol (10 μM) caused a significant activation of the PMRS (41%) and AFR reductase (30%) over (basal level) the control; the effect of resveratrol was concentration-dependent. The electron donating ability of resveratrol is slightly less than that observed with quercetin. The role of resveratrol in activating the erythrocyte PMRS and AFR reductase may assume significance in all disease conditions in which there is a decrease in plasma antioxidant potential.     

Contradistinction between doxorubicin and epirubicin: in-vitro interaction with blood components.

The molecular structure and anti-tumour activity of doxorubicin and epirubicin are similar. However, the incidence of their cardiotoxicity occurs at different cumulative dose concentrations. The purpose of this study was to investigate the in-vitro interaction of these two drugs with different blood components, namely intact erythrocytes, haemoglobin and erythrocyte ghosts. Plasma protein binding was also evaluated. The intended goal was to identify the most relevant samples among total blood, plasma or blood cells for pharmacokinetic analysis. The methodology involved the incubation of each of the blood components (the intact erythrocytes, erythrocyte ghosts, haemoglobin and plasma proteins) at physiological pH and temperature with different concentrations of each drug, followed by measurement by HPLC and fluorometry at excitation and emission wavelengths of 480 and 580 nm, respectively. The results indicated that the binding of doxorubicin and epirubicin to plasma proteins, erythrocyte ghosts and intact erythrocytes was essentially the same. However, the binding of both compounds to intact erythrocytes was significantly different from erythrocyte ghosts, which indicates that haemoglobin plays an important role in the binding to and uptake by erythrocytes. The isotherms of binding to haemoglobin revealed that the maximum binding of doxorubicin was approximately 0.42 microg mg(-1) haemoglobin; for epirubicin this value was ten times greater than for doxorubicin. The Scatchard plot of binding of both drugs to haemoglobin exhibited two distinct binding sites for each drug. The constant of association of high affinity and low capacity binding sites was significantly greater for epirubicin, whereas the constant of association of low affinity and high capacity binding sites was significantly higher for doxorubicin. The number of high affinity binding sites per mg of haemoglobin was estimated to be 0.072 for doxorubcin and 0.030 for epirubicin. The number of low affinity binding sites was significantly greater for epirubicin (1.963) than for doxorubicin (0.305). Since the combined number of binding sites for epirubicin was more than doxorubicin, and the total uptake by erythrocytes remained the same for both drugs, it was concluded that epirubicin, being a more lipophilic compound, may diffuse more freely into the cells. Therefore, it binds more to haemoglobin, whereas doxorubicin remains more adsorbed on the surface of the cells due to its self-association property. It was concluded that the interaction of both drugs with erythrocytes, although it appears to be similar, is significantly different due to the interaction with haemoglobin. The difference in this interaction is expected to influence the disposition of both drugs in-vivo.