Oxidative Hemolysis Of Human Erythrocytes Research Articles

Thymoquinone enhances the antioxidant and anticancer activity of Lebanese propolis.

Reactive oxygen species (ROS) are produced by multiple cellular processes and are maintained at optimal levels in normal cells by endogenous antioxidants. In recent years, the search for potential exogenous antioxidants from dietary sources has gained considerable attention to eliminate excess ROS that is associated with oxidative stress related diseases including cancer. Propolis, a resinous honeybee product, has been shown to have protective effects against oxidative stress and anticancer effects against several types of neoplasms. To investigate the antioxidant and anticancer potential of Lebanese propolis when applied alone or in combination with the promising anticancer compound Thymoquinone (TQ) the main constituent of Nigella sativa essential oil. Crude extracts of Lebanese propolis collected from two locations, Rashaya and Akkar-Danniyeh, were prepared in methanol and the total phenolic content was determined by Folin-Ciocalteu method. The antioxidant activity was assessed by the ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical and to inhibit H2O2-induced oxidative hemolysis of human erythrocytes. The anticancer activity was evaluated by [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide] MTT assay against HCT-116 human colorectal cancer cells and MDA-MB-231 human breast cancer cells. The total phenolic content of propolis extract from Rashaya and Akkar-Danniyeh were 56.81 µg and 83.503 µg of gallic acid equivalent /mg of propolis, respectively. Both natural agents exhibited strong antioxidant activities as evidenced by their ability to scavenge DPPH free radical and to protect erythrocytes against H2O2-induced hemolysis. They also dose-dependently decreased the viability of both cancer cell lines. The IC50 value of each of propolis extract from Rashaya and Akkar-Danniyeh or TQ was 22.3, 61.7, 40.44 µg/mL for breast cancer cells at 72 h and 33.3, 50.9, 33.5 µg/mL for colorectal cancer cells at the same time point, respectively. Importantly, the inhibitory effects of propolis on DPPH radicals and cancer cell viability were achieved at half its concentration when combined with TQ. Our results indicate that Lebanese propolis extract has antioxidant and anticancer potential and its combination with TQ could possibly prevent ROS- mediated diseases.

Protective Effects of Auraptene against Free Radical-Induced Erythrocytes Damage.

Auraptene is the most abundant natural prenyloxycoumarin. Recent studies have shown that it has multiple biological and therapeutic properties, including antioxidant properties. Erythrocytes are constantly subjected to oxidative damage that can affect proteins and lipids within the erythrocyte membrane and lead to some hemoglobinopathies. Due to the lack of sufficient information about the antioxidant effects of auraptene on erythrocytes, this study intended to evaluate the potential of this compound in protecting radical-induced erythrocytes damages. The antioxidant activity of auraptene was measured based on DPPH and FRAP assays. Notably, oxidative hemolysis of human erythrocytes was used as a model to study the ability of auraptene to protect biological membranes from free radical-induced damage. Also, the effects of auraptene in different concentrations (25-400 µM) on AAPH-induced lipid/protein peroxidation, glutathione (GSH) content and morphological changes of erythrocytes were determined. Oxidative hemolysis and lipid/protein peroxidation of erythrocytes were significantly suppressed by auraptene in a time and concentration-dependent manner. Auraptene prevented the depletion of the cytosolic antioxidant GSH in erythrocytes. Furthermore, it inhibited lipid and protein peroxidation in a time and concentration-dependent manner. Likewise, FESEM results demonstrated that auraptene reduced AAPH-induced morphological changes in erythrocytes. Auraptene efficiently protects human erythrocytes against free radicals. Therefore, it can be a potent candidate for treating oxidative stress-related diseases.

Cyanidin-3-rutinoside reduces insulin fibrillation and attenuates insulin fibrils-induced oxidative hemolysis of human erythrocytes

Insulin is able to form amyloid-like fibrils, a misfolding process by which insulin molecules interact with each other to form aggregates and pathological amyloid deposition. Inhibition of amyloid aggregation using natural products is proposed as a new strategy to prohibit the development of amyloid diseases. Herein, we demonstrated the inhibitory effect of cyanidin-3-rutinoside (C3R), a natural anthocyanin with multiple biological activities, against insulin amyloid fibrillation. The results showed that increased insulin concentration resulted in faster growth and higher amounts of insulin fibrils. C3R (10.6–170μM) concentration dependently decreased insulin fibril growth and increased the duration of lag time of insulin fibril formation. Moreover, C3R directly decreased the secondary structure transition from α-helix to β-sheet structure. C3R (0.31–5μM) attenuated insulin fibrils-induced oxidative hemolysis of human erythrocytes in a concentration-dependent manner. Moreover, C3R reduced insulin fibrils-induced erythrocyte membrane disruption through the inhibition of reactive oxygen species (ROS) generation. The findings also suggest that C3R reduced fibrils-induced membrane lipid peroxidation by maintaining the catalase activity and oxidized/reduced glutathione content (GSH/GSSH) in erythrocytes. These findings suggest that C3R may serve as a potential inhibitory agent against amyloid fibril formation and insulin fibrils-induced oxidative hemolysis.

Physicochemical Characterization, Microbiological Quality and Safety, and Pharmacological Potential of Hancornia speciosa Gomes

Hancornia speciosa Gomes is a fruit tree, commonly known as the mangaba tree, which is widespread throughout Brazil. The leaves of this plant are used in traditional medicine for medicinal purposes. Thus, the objective of this study was to perform a physicochemical characterization, identify the lipophilic antioxidants and fatty acids, and determine the microbiological quality and safety of H. speciosa leaves. In addition, the antioxidant, antimutagenic, and inhibitory activities of the ethanolic extract of H. speciosa leaves (EEHS) against enzymes related to neurodegenerative diseases, inflammation, obesity, and diabetes were investigated. Furthermore, this study aimed at assessing the in vivo effects of the EEHS on the glycemia of normoglycemic and diabetic Wistar rats. Physicochemical characterization was performed by colorimetry and gas-liquid chromatography with flame ionization detection (GC-FID). The total number of colonies of aerobic mesophiles, molds, and yeasts was determined. The total coliforms and Escherichia coli were counted using the SimPlates kit, and sulphite-reducing Clostridium spores were quantified using the sulphite-polymyxin-sulfadiazine agar method. Salmonella spp. were detected using the 1-2 Test. The antioxidant activity of the EEHS was measured by its inhibition of 2,2′-azobis(2-amidinopropane) dihydrochloride- (AAPH-) induced oxidative hemolysis of human erythrocytes. The antimutagenic activity was determined using the Ames test. The acetylcholinesterase, butyrylcholinesterase, tyrosinase, hyaluronidase, lipase, α-amylase, and α-glycosidase enzyme-inhibiting activities were assessed and compared with commercial controls. The in vivo effects of the EEHS were assessed using the oral glucose tolerance test in normoglycemic Wistar rats and measuring the blood glucose levels in diabetic rats. The results demonstrated physical-chemical parameters of microbiological quality and safety in the leaves of H. speciosa, as well as antioxidant and antimutagenic activities and inhibition of enzymes related to neurodegenerative diseases, inflammation, obesity, and diabetes. In in vivo assays, it was shown that the normoglycemic rats challenged with glucose overload show significantly decreased blood glucose levels when treated with the EEHS. Taken together, the results ensure the microbiological quality and safety as well as showing the contents of carotenoids and polyunsaturated fatty acids of H. speciosa leaves. Additionally, the antioxidant, antimutagenic, anti-inflammatory, anti-Alzheimer's disease, anti-Parkinson's disease, antiobesity, and antihyperglycemic activities of the EEHS were demonstrated.

Antrodia salmonea in submerged culture exhibits antioxidant activities in vitro and protects human erythrocytes and low-density lipoproteins from oxidative modification

Antrodia salmonea is well known in Taiwan as a beneficial mushroom. In the present study, we investigated the antioxidant activity of whole fermented broth (AS), filtrate (ASF), and mycelia (ASM) of A. salmonea using different antioxidant models. Furthermore, the effect of A. salmonea on AAPH-induced oxidative hemolysis of human erythrocytes and CuSO4-induced oxidative modification of human low-density lipoproteins (LDLs) was examined. We found that the AS, ASF, and ASM possess effective antioxidant activity against various oxidative systems including superoxide anion scavenging, reducing power, metal chelation, and DPPH radical scavenging. Further, AAPH-induced oxidative hemolysis in erythrocytes was prevented by AS, ASF, and ASM. Notably, AS, ASF, and ASM appear to possess powerful antioxidant activities against CuSO4-induced oxidative modification of LDL as assessed by malondialdehyde (MDA) formation, cholesterol degradation, and the relative electrophoretic mobility of oxidized LDL. It is noteworthy that AS had comparatively strong antioxidant ability compared to ASF or ASM, which is well correlated with the content of their total polyphenols. Thus, A. salmonea may exert antioxidant properties and offer protection from atherogenesis.

Synthesis and Evaluation of Antioxidant Properties of Novel 1,2,4‐Triazole‐Based Schiff Base Heterocycles

A series of 1,2,4-triazole-based Schiff base heterocyclic compounds (5a-f and 8a-i) and phenethylamines (7a-h) were synthesized and evaluated for antioxidant properties by free-radical scavenging, anti-hemolytic activity, lipid peroxidation, and their protective effects against DNA oxidative damage. Compounds 7c, 7d, 7h, 8b, and 8i showed promising DPPH(•) radical scavenging activity with the level of inhibition between 86.8% and 94%. Compounds 8a, 8b, 8d, 8g, and 8i were effective against the oxidative hemolysis of human erythrocytes and lipid peroxidation, in a dose-dependent manner, with IC50 values in the range of 55.7-80.7 and 53.2-81.2 µg/mL, respectively. Compounds 8a and 8b were effective against oxidative damage on erythrocyte ghost membrane proteins, and 8g and 8i were able to protect against DNA oxidative damage.

Dracaena draco L. fruit: Phytochemical and antioxidant activity assessment

The present study reports for the first time the metabolite profile and antioxidant activity of aqueous extract obtained from Dracaena draco L. fruit. Volatiles profile was determined by HS-SPME/GC-IT-MS, with 9 compounds being identified, distributed by several distinct chemical classes: 1 alcohol, 3 aldehydes, 2 carotenoid derivatives, and 3 terpenic compounds. Aldehydes constituted the most abundant class in this exotic berry, representing 59% of total identified volatile compounds. Phenolics profile was determined by HPLC/DAD and 5 constituents were identified: 5-O-caffeoylquinic, 3,5-O-dicaffeoylquinic, ferulic and sinapic acids, and quercetin-3-O-rutinoside. The major phenolic compound is quercetin-3-O-rutinoside, comprising 42% of the total phenolic content. Organic acids composition was also characterized, by HPLC-UV, and oxalic, citric, l-ascorbic, malic, quinic and shikimic acids were determined. The most abundant is quinic acid, representing 39% of the total organic acid content. The antioxidant potential of this matrix was assessed by (i) reducing power of Fe3+/ferricyanide complex, (ii) scavenging effect on DPPH free radicals, and (iii) ability to inhibit the 2,2´-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis in human erythrocytes. Strawberry (Fragaria×ananassa Duch. cv. Camarosa) extract was used for comparison purposes. All assay models showed remarkable concentration dependent antioxidant activity, reducing power and radical scavenging efficiency for D. draco fruit, being invariably higher than that of strawberry extract. This is the first report showing that D. draco fruit is a promising new antioxidant agent.

Human cancer cell antiproliferative and antioxidant activities of Juglans regia L.

Several studies suggest that regular consumption of nuts, mostly walnuts, may have beneficial effects against oxidative stress mediated diseases such as cardiovascular disease and cancer. Walnuts contain several phenolic compounds which are thought to contribute to their biological properties. The present study reports the total phenolic contents and antioxidant properties of methanolic and petroleum ether extracts obtained from walnut ( Juglans regia L.) seed, green husk and leaf. The total phenolic contents were determined by the Folin–Ciocalteu method and the antioxidant activities assessed by the ability to quench the stable free radical 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes. Methanolic seed extract presented the highest total phenolic content (116 mg GAE/g of extract) and DPPH scavenging activity (EC 50 of 0.143 mg/mL), followed by leaf and green husk. In petroleum ether extracts, antioxidant action was much lower or absent. Under the oxidative action of AAPH, all methanolic extracts significantly protected the erythrocyte membrane from hemolysis in a time- and concentration-dependent manner, although leaf extract inhibitory efficiency was much stronger (IC 50 of 0.060 mg/mL) than that observed for green husks and seeds (IC 50 of 0.127 and 0.121 mg/mL, respectively). Walnut methanolic extracts were also assayed for their antiproliferative effectiveness using human renal cancer cell lines A-498 and 769-P and the colon cancer cell line Caco-2. All extracts showed concentration-dependent growth inhibition toward human kidney and colon cancer cells. Concerning A-498 renal cancer cells, all extracts exhibited similar growth inhibition activity (IC 50 values between 0.226 and 0.291 mg/mL), while for both 769-P renal and Caco-2 colon cancer cells, walnut leaf extract showed a higher antiproliferative efficiency (IC 50 values of 0.352 and 0.229 mg/mL, respectively) than green husk or seed extracts. The results obtained herein strongly indicate that walnut tree constitute an excellent source of effective natural antioxidants and chemopreventive agents.

Protective effect of quince (Cydonia oblonga Miller) fruit against oxidative hemolysis of human erythrocytes

The aim of this study was to determine the phenolic content and evaluate the antioxidant activity of quince (Cydonia oblonga) fruit. For this purpose, fruits were separated into pulps, peels and seeds and methanolic extracts were prepared. The phenolic profiles were determined by HPLC/UV and antioxidant properties were studied for their ability to quench the stable free radical 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes.The main phenolic compounds were 5-O-caffeoylquinic acid for pulp and peel (57% and 29%, respectively) and stellarin-2 for seed (18%). Total phenolics content was 2.5, 6.3 and 0.4g/kg of methanolic extract for pulp, peel and seed, respectively. Pulp and peel extracts showed similar DPPH free radical scavenging activities (EC50 of 0.6 and 0.8mg/ml, respectively), while seed extract presented much lower antioxidant potential (EC50 of 12.2mg/ml). Under the oxidative action of AAPH, pulp and peel extracts showed significant protection of the erythrocyte membrane from hemolysis, in a time- and concentration-dependent manner. Seed extracts by themselves induced extensive hemolysis. These results indicate higher antioxidant activity for certain parts of quince fruit, namely pulp and peel, that may therefore represent accessible sources of natural antioxidants with potential application in nutritional/pharmaceutical fields, as preventive or therapeutic agents in diseases in which free radicals are implicated.

Evaluation of free radical-scavenging and antihemolytic activities of quince ( Cydonia oblonga) leaf: A comparative study with green tea ( Camellia sinensis)

This study aimed to determine the phenolic profile and to investigate the antioxidant potential of quince ( Cydonia oblonga) leaf, comparing it with green tea ( Camellia sinensis). For these purposes, methanolic extracts were prepared and phenolics content of quince leaf was determined by HPLC/UV. The antioxidant properties were assessed by Folin–Ciocalteu reducing capacity assay and by the ability to quench the stable free radical 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes. 5- O-Caffeoylquinic acid was found to be the major phenolic compound in quince leaf extract. Quince leaf exhibited a significantly higher reducing power than green tea (mean value of 227.8 ± 34.9 and 112.5 ± 1.5 g/kg dry leaf, respectively). Quince leaf extracts showed similar DPPH radical-scavenging activities (EC 50 mean value of 21.6 ± 3.5 μg/ml) but significantly lower than that presented by green tea extract (EC 50 mean value of 12.7 ± 0.1 μg/ml). Under the oxidative action of AAPH, quince leaf methanolic extract significantly protected the erythrocyte membrane from hemolysis in a similar manner to that found for green tea (IC 50 mean value of 30.7 ± 6.7 and 24.3 ± 9.6 μg/ml, respectively, P > 0.05). These results point that quince leaf may have application as preventive or therapeutic agent in diseases in which free radicals are involved.

Protective effect of some herbal medicine against oxidative hemolysis of human erythrocytes

Protective effect of some herbal medicine against oxidative hemolysis of human erythrocytes

Protective effects of crataegus curvisepala extracts against oxidative hemolysis of human erythrocytes

Protective effects of crataegus curvisepala extracts against oxidative hemolysis of human erythrocytes

Protective effect of some herbal medicine against oxidative hemolysis of human erythrocytes

Protective effect of some herbal medicine against oxidative hemolysis of human erythrocytes