AAPH-induced Oxidation Research Articles

The Impact of AAPH-Induced Oxidation on the Functional and Structural Properties, and Proteomics of Arachin.

The aim of this study was to investigate the effect of 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidation on the functional, structural properties and proteomic information of arachin. The results showed that moderate oxidation improved the water/oil holding capacity of proteins and increased the emulsifying stability, while excessive oxidation increased the carbonyl content, reduced the thiol content, altered the structure and thermal stability, and reduced most of the physicochemical properties. Through LC-QE-MS analysis, it was observed that oxidation leads to various modifications in arachin, including carbamylation, oxidation, and reduction, among others. In addition, 15 differentially expressed proteins were identified. Through gene ontology (GO) analysis, these proteins primarily affected the cellular and metabolic processes in the biological process category. Further Kyoto encyclopedia of genes and genomes (KEGG) analysis revealed that the "proteasome; protein processing in the endoplasmic reticulum (PPER)" pathway was the most significantly enriched signaling pathway during the oxidation process of arachin. In conclusion, this study demonstrated that AAPH-induced oxidation can alter the conformation and proteome of arachin, thereby affecting its corresponding functional properties. The findings of this study can potentially serve as a theoretical basis and foundational reference for the management of peanut processing and storage.

Peroxyl radical scavenging activity measurement of antioxidants using histidine-stabilized and glutathione-capped fluorescent gold nanoclusters

A fluorescent gold nanocluster was used for determining peroxyl radical scavenging activity of antioxidants. Histidine was used as a green reducing and protective agent, and glutathione (GSH) enhanced the fluorescence intensity of histidine-stabilized gold nanoclusters (AuNCs) by ligand exchange process. When AAPH-induced oxidation of GSH occurred, the initial fluorescence intensity of GSH-capped AuNCs (λex = 450 nm λem = 502 nm) was decreased with static quenching. The decline of fluorescence intensity of the GSH-capped AuNCs upon peroxyl radical attack is diminished with the addition of antioxidants to the reaction medium, the difference in fluorescence intensity being related to peroxyl radical scavenging activity of antioxidants. The 50 % inhibitive concentration of related antioxidant compounds were determined and compared to those of crocin bleaching assay. Inhibition % of sage (Salvia officinalis L.) and green tea (Camellia sinensis) infusions against peroxyl radicals were investigated. The proposed assay can be used for simple and selective estimation of the peroxyl radical scavenging activity in complex matrices, as histidine-stabilized GSH-capped AuNCs were selective toward peroxyl radicals, not affected by other ROS at the studied concentrations.

Antioxidant properties and free radicals scavenging activities of pomegranate (Punica granatum L.) peels: An in-vitro study

Fruit peels often contain more bioactive substances than the edible component of the fruit. As a result, the current study was designed to use a variety of in-vitro assays to investigate the antioxidant capacity of phenolic compounds found in pomegranate (Punica granatum L.) peel. The IC50 values for DPPH free radical scavenging activity, ABTS assay, and lipid peroxidation inhibition activity were as low as 0.1, whereas the same for hydroxyl radical scavenging activity was 0.6 μg/ml. The nitric oxide scavenging activity had the lowest IC50 value (0.02 μg/ml). At a concentration of 0.21 μg/ml, phenolics scavenged ∼60% of DPPH radicals, while at a phenolic content of 98.9 μg/ml, the extract chelated 60% of the iron metal. The RP0.5AU value of total reducing power was 0.37 μg/ml. Total antioxidant activity (AO0.5AU value of 4.24 μg/ml) was significantly lower than the standard (8.42 g/ml). At a dose of 0.09 μg/ml, phenolics reduced lipid peroxidation by 53% in an egg homogenate model. Gel electrophoresis indicated that the extracts with phenolic content of 20 and 40 μg virtually protected the BSA against AAPH-induced oxidation. Overall, various experiments showed that pomegranate peel extract has significant antioxidant activity and supports its use as a source of natural antioxidants to combat autoxidation-induced pathologies or diseases and use in the health food/nutraceutical/pharmaceutical industry for a variety of applications.

Comparison of Antioxidants: The Limited Correlation between Various Assays of Antioxidant Activity.

The inhibitory effects a range of synthetic and natural antioxidants on lipid peroxidation of egg yolk and erythrocyte membranes induced by a free radical generator 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH) was compared, with significant differences being found between both systems. When the protection by selected antioxidants against the effects of AAPH on erythrocytes (hemolysis, oxidation of hemoglobin and glutathione (GSH) and generation of reactive oxygen species (ROS)) was studied, most antioxidants were protective, but in some tests (oxidation of hemoglobin and GSH) some acted as prooxidants, inducing oxidation in the absence of AAPH and enhancing the AAPH-induced oxidation. These results demonstrate a diversified action of antioxidants in different systems and point to a need for careful extrapolation of any conclusions drawn from one parameter or experimental system to another.

Nitrone-Trolox conjugate as an inhibitor of lipid oxidation: Towards synergistic antioxidant effects

Free radical scavengers like α-phenyl-N-tert-butylnitrone (PBN) and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) have been widely used as protective agents in various biomimetic and biological models. A series of three amphiphilic Trolox and PBN derivatives have been designed by adding to those molecules a perfluorinated chain as well as a sugar group in order to render them amphiphilic. In this work, we have studied the interactions between these derivatives and lipid membranes to understand how they influence their ability to prevent membrane lipid oxidation. We showed the derivatives better inhibited the AAPH-induced oxidation of 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLiPC) small unilamellar vesicles (SUVs) than the parent compounds. One of the derivatives, bearing both PBN and Trolox moieties on the same fluorinated carrier, exhibited a synergistic antioxidant effect by delaying the oxidation process. We next investigated the ability of the derivatives to interact with DLiPC membranes in order to better understand the differences observed regarding the antioxidant properties. Surface tension and fluorescence spectroscopy experiments revealed the derivatives exhibited the ability to form monolayers at the air/water interface and spontaneously penetrated lipid membranes, underlying pronounced hydrophobic properties in comparison to the parent compounds. We observed a correlation between the hydrophobic properties, the depth of penetration and the antioxidant properties and showed that the location of these derivatives in the membrane is a key parameter to rationalize their antioxidant efficiency. Molecular dynamics (MD) simulations supported the understanding of the mechanism of action, highlighting various key physical-chemical descriptors.

A Novel Nitrone-Trolox Conjugate Inhibits Membrane Lipid Oxidation Through Synergistic Antioxidant Effects

Polyunsaturated fatty acids are particularly sensitive to the deleterious effects of reactive oxygen species (ROS) as ROS can trigger their degradation through a chemical reaction called lipid peroxidation. Lipid oxidation has been reported to be involved in the early stages of several diseases as well as the deterioration of food supplies. To prevent the deleterious effects of ROS, water soluble free radical scavengers like α-phenyl-N-tert-butylnitrone (PBN) and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) have been widely used as protective agents in various biomimetic and biological models. In order to increase the hydrophobicity and the ability of these compounds to interact with membranes, three amphiphilic Trolox and PBN derivatives were recently designed by adding a perfluorinated chain to Trolox and PBN. A sugar moiety was added as well to ensure water solubility. In the present work, we studied the physical-chemical properties of the derivatives and related their antioxidant properties to their ability to interact with lipid membranes. We first investigated their ability to inhibit the AAPH-induced oxidation of 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC) liposomes as well as their ability to interact with DLPC membranes. Results evidenced that the three derivatives are efficient to prevent lipid oxidation and that the location in the hydrophobic core of the membrane is a key parameter in the antioxidant efficiency. The derivative bearing both PBN and Trolox moieties on the same fluorinated carrier exhibited a synergistic antioxidant effect by delaying the oxidation process and by scavenging free radicals. Molecular dynamics simulations supported the understanding of the mechanism of action, highlighting various key physical-chemical descriptors.

A novel colorimetric sensor for measuring hydroperoxide content and peroxyl radical scavenging activity using starch-stabilized gold nanoparticles

A novel colorimetric nanosensor was developed for evaluating peroxyl radical scavenging activity of phenolic antioxidants and for the detection of hydroperoxides formed during AAPH-induced oxidation of linoleic acid emulsions. Starch was used as a green reduction/stabilization agent for gold nanoparticles (AuNPs) synthesis in alkaline medium. When tert-butyl hydroperoxide (tert-BHP) was incubated with an excess of iodide ions in a 37 °C water bath for 90 min, triodide (I3–) was formed in an amount equivalent to tert-BHP concentration. Upon the addition of starch-stabilized gold nanoparticles (ss-AuNPs) solution to the incubation mixture, triiodide ions were rapidly adsorbed on the surface of AuNPs and caused their aggregation. A concomitant red shift (from 525 nm to 563 nm) of surface plasmon resonance (SPR) absorption of the nanoparticles was observed, absorbance linearly increasing with aqueous tert-BHP concentration. The method provided an LOD of 39 µM for tert-BHP, and was validated through linearity, precision and accuracy. The concentration of hydroperoxides estimated in linoleic acid peroxidation correlated well with those found by the reference ferric thiocyanate assay. Peroxyl radical scavenger antioxidants decreased the red-shifted SPR absorption of aggregated ss-AuNPs, thereby enabling an indirect estimation of antioxidant activity. This AuNPs-based colorimetric sensor is the first of its kind to directly determine peroxyl radical scavenging activity of polyphenols. The half-maximal inhibitive concentrations (IC50) of selected antioxidant compounds were calculated by utilizing the decrease in absorbance with increasing concentration of scavengers, and compared to those of classical oxygen radical absorbance capacity (ORAC) assay. The proposed nanosensor was superior over FL-based ORAC in determining the peroxyl radical scavenging activity of the lipophilic antioxidant α-tocopherol. The percentage scavenging of real samples such as green tea infusion and synthetic serum were determined. The proposed assay can be used for estimating the peroxyl scavenging of various food and biological samples in terms of its low cost, ease of use and compatibility.

Dual Effect of Histidine on Polysorbate 20 Stability: Mechanistic Studies.

L-Histidine (L-His) and polysorbate 20 (PS20) are two excipients frequently included in parenteral products to stabilize biotherapeutics. The objective of the current work was to investigate the impact of L-His on PS20 stability in aqueous solutions when subjected to forced oxidation and accelerated stability testing. The stability of PS20 in L-His buffer was compared with that in acetate buffer. Forced oxidation of PS20 in these two buffer systems was initiated by a free radical generator, 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH), while accelerated stability tests were carried out at 40°C. Ultra-performance liquid chromatography mass spectrometry was utilized to monitor intact PS20 and to analyze degradation products. Our results demonstrate a dual effect of L-His on PS20 stability. During exposure to AAPH, L-His protects PS20 from oxidation. Stable isotope labeling of L-His with 13C was employed for mechanistic investigations. The protection of L-His was abrogated when acetate was added to L-His buffer, implying that the anti-oxidative activity of L-His may be compromised by specific counter ions. The replacement of L-His by various His derivatives led to significant changes in the protection of PS20 against AAPH-induced degradation. In contrast to forced degradation, the addition of L-His promoted oxidative PS20 degradation during accelerated storage at 40°C in solution, generating mainly short chain POE-laurates. L-His exhibits a dual effect on the stability profile of PS20, protecting against AAPH-induced oxidation but promoting oxidative degradation during accelerated stability testing.

Attenuation of oxidative stress and artificial wound closure in C2C12 myoblasts induced by sequential extracts of Boerhavia diffusa.

Whole plants of Boerhavia diffusa L. are widely used medicine in Ghana and other tropical countries, for the treatment of wounds and other ailments. The aim of the study was to determine the ability of sequential extracts of B.diffusa to influence oxidation and wound closure in myoblast cells invitro. Sequential extracts were prepared from the whole plant using four solvents of increasing polarity (hexane, ethyl acetate, methanol and water). Cytotoxicity was determined using the sulforhodamine B staining assay, phase-contrast microscopy, plasDIC microscopy and live-dead staining. Extracts were tested for their ability to reduce 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidation and mediate cell migration after artificial wound generation in C2C12 myoblast cells using the scratch wound assay. All extracts indicated negligible cytotoxicity (IC50 >100μg/ml), and microscopic evaluation showed no difference from negative controls. AAPH induced a 2.87-fold increase in reactive oxygen species compared to the negative control. Pretreatment with 100μg/ml of the extracts reduced AAPH-induced oxidation to 1.70-fold of the untreated controls (P<0.001). Wound closures in the methanol and water extract treatments were 18.08% and 20.76% higher than the negative control, respectively (P<0.01). These findings indicate that the hexane, methanol and water extracts of B.diffusa whole plant promote artificial wound healing and protection against oxidation invitro and therefore warrant further research into its mechanisms of wound healing.

Antioxidant properties of catechins: Comparison with other antioxidants

Antioxidant properties of five catechins and five other flavonoids were compared with several other natural and synthetic compounds and related to glutathione and ascorbate as key endogenous antioxidants in several in vitro tests and assays involving erythrocytes. Catechins showed the highest ABTS-scavenging capacity, the highest stoichiometry of Fe3+ reduction in the FRAP assay and belonged to the most efficient compounds in protection against SIN-1 induced oxidation of dihydrorhodamine 123, AAPH-induced fluorescein bleaching and hypochlorite-induced fluorescein bleaching. Glutathione and ascorbate were less effective. (+)-catechin and (−)-epicatechin were the most effective compounds in protection against AAPH-induced erythrocyte hemolysis while (−)-epicatechin gallate, (−)-epigallocatechin gallate and (−)-epigallocatechin protected at lowest concentrations against hypochlorite-induced hemolysis. Catechins [(−)-epigallocatechin gallate and (−)-epicatechin gallate)] were most efficient in the inhibition of AAPH-induced oxidation of 2′7′-dichlorodihydroflurescein contained inside erythrocytes. Excellent antioxidant properties of catechins and other flavonoids make them ideal candidates for nanoformulations to be used in antioxidant therapy.

Tetrahydropyrrolization of Resveratrol and Other Stilbenes Improves Inhibitory Effects on DNA Oxidation

The inhibitory effect of resveratrol on DNA oxidation caused by 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH) was found to be enhanced if the C=C bond in resveratrol was converted into tetrahydropyrrole by reaction with azomethine ylide (CH2 =N(+) (CH3 )CH2 (-) ). This encouraged us to explore whether the inhibitory activities of other stilbenes could also be increased by the same method. We found that the inhibitory effects of the tetrahydropyrrole derivatives on AAPH-induced oxidation of DNA were higher than those of the corresponding stilbenes, because the tetrahydropyrrole motif can provide hydrogen atoms to be abstracted by radicals. Therefore, the tetrahydropyrrolization offered an advantage for enhancing the antioxidant effects of stilbenes. Notably, (CH3 )3 SiCH2 N(CH3 )CH2 OCH3 (in the presence of CF3 COOH) and (CH3 )3 NO (in the presence of LiN(iPr)2 ) can be used to generate azomethine ylide for the tetrahydropyrrolization of stilbenes containing electron-withdrawing and -donating groups, respectively.

In-vitro Evaluation of Protective Effects on DNA Damage and Antioxidative Activities of Ilex Spinigera Loes. Extracts.

Ilex spinigera (Aquifoliaceae) plant is an evergreen tree or shrub with thick glossy dark green leaves and red fruits. This plant has medicinal properties and has been used traditionally in northern Iran for malaria treatment. The aim of this work is to evaluate the antioxidative activities and the inhibitory effect of I. spinigera on the oxidation of DNA. We have found no reports about the popular use of I. spinigera in terms of its chemistry and biology. In this study we report the antioxidant activity of I. spinigera extracts for the first time. Water, ethanol and methanol were used as extraction solvents. Various experimental models including iron (III) reducing power, total antioxidant capacity, DPPH radical scavenging activity, PAB assay and in-vitro inhibition of AAPH-induced oxidation of DNA were used for characterization of antioxidant activity of the extracts. The three extracts showed varying degrees of efficacy in each assay in a dose-dependent manner. The aqueous extract with the highest content of total phenolics, was the most potent antioxidant in all assays except in DPPH assay. The methanol extract with the highest amount of total flavonoids was the potent scavenger of DPPH radical with an IC50 value of 102.22 ± 0.001 μg/mL. Aqueous extract of I. spinigera also showed the protective effect on DNA damage-induced by AAPH. According to our results, I. spinigera leaves extract have the potential for chemoprotective studies.

Activity of coumarin–oxadiazole-appended phenol in inhibiting DNA oxidation and scavenging radical

Fourteen coumarin–oxadiazole-appended phenols were synthesized, while halogen atom, methoxyl, and hydroxyl groups acted as the functional groups for testing the ability to trap 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS+) and to inhibit the oxidation of DNA caused by 2,2′-azobis(2-amidinopropane hydrochloride) (AAPH) and Cu2+/glutathione (GSH). The coumarin–oxadiazole-appended phenols containing the hydroxyl group were able to inhibit AAPH-induced oxidation of DNA and to quench ABTS+, and the antioxidant effectiveness depended upon the number of hydroxyl groups. Moreover, the coumarin–oxadiazole-appended phenols used herein were able to inhibit Cu2+/GSH-induced oxidation of DNA, and the inhibitory effect generated by bromide was similar to that of the hydroxyl group. In particular, the antioxidant effectiveness of para-bromide at the benzene ring even approached to that of Trolox.

Interaction of α-Hexylcinnamaldehyde with a Biomembrane Model: A Possible MDR Reversal Mechanism.

The ability of the naturally derived compound α-hexylcinnamaldehyde (1) to interact with biomembranes and to modulate their permeability has been investigated as a strategy to reverse multidrug resistance (MDR) in cancer cells. Dimyristoylphosphatidylcholine (DMPC) multilamellar vesicles (MLVs) were used as biomembrane models, and differential scanning calorimetry was applied to measure the effect of 1 on the thermotropic behavior of DMPC MLVs. The effect of an aqueous medium or a lipid carrier on the uptake of 1 by the biomembrane was also characterized. Furthermore, taking into account that MDR is strictly regulated by redox signaling, the pro-oxidant and/or antioxidant effects of 1 were evaluated by the crocin-bleaching assay, in both hydrophilic and lipophilic environments. Compound 1 was uniformly distributed in the phospholipid bilayers and deeply interacted with DMPC MLVs, intercalating among the phospholipid acyl chains and thus decreasing their cooperativity. The lipophilic medium allowed the absorption of 1 into the phospholipid membrane. In the crocin-bleaching assay, the substance produced no pro-oxidant effects in both hydrophilic and lipophilic environments; conversely, a significant inhibition of AAPH-induced oxidation was exerted in hydrophilic medium. These results suggest a possible role of 1 as a chemopreventive and chemosensitizing agent for fighting cancer.

Coumarin-fused coumarin: antioxidant story from N,N-dimethylamino and hydroxyl groups.

Two coumarin skeletons can form chromeno[3,4-c]chromene-6,7-dione by sharing with the C ═ C in lactone. The aim of the present work was to explore the antioxidant effectiveness of the coumarin-fused coumarin via six synthetic compounds containing hydroxyl and N,N-dimethylamino as the functional groups. The abilities to quench 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS(+•)), 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH), and galvinoxyl radical revealed that the rate constant for scavenging radicals was related to the amount of hydroxyl group in the scaffold of coumarin-fused coumarin. But coumarin-fused coumarin was able to inhibit DNA oxidations caused by (•)OH, Cu(2+)/glutathione (GSH), and 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH) even in the absence of hydroxyl group. In particular, a hydroxyl and an N,N-dimethylamino group locating at different benzene rings increased the inhibitory effect of coumarin-fused coumarin on AAPH-induced oxidation of DNA about 3 times higher than a single hydroxyl group, whereas N,N-dimethylamino-substituted coumarin-fused coumarin possessed high activity toward (•)OH-induced oxidation of DNA without the hydroxyl group contained. Therefore, the hydroxyl group together with N,N-dimethylamino group may be a novel combination for the design of coumarin-fused heterocyclic antioxidants.

Ferrocenyl-appended aurone and flavone: which possesses higher inhibitory effects on DNA oxidation and radicals?

The aim of the present work was to compare the antioxidative effect of the ferrocenyl-appended aurone with that of ferrocenyl-appended flavone; therefore, nine aurones together with the flavone-type analogues were synthesized by using chalcone as the reactant. The radical-scavenging property was evaluated by reacting with the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS(+·)), 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH), and galvinoxyl radical, respectively. The cytotoxicity was estimated by inhibiting 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH)-induced oxidation of DNA. It was found that the introduction of the ferrocenyl group remarkably increased the radical-scavenging activities of aurone and flavone. Especially, the ferrocenyl group in flavones can quench radicals even in the absence of the phenolic hydroxyl group, while ferrocenyl-appended aurones can efficiently protect DNA against AAPH-induced oxidation. Therefore, the antioxidative effect was generated by the ferrocenyl group and enhanced by the electron-donating group attaching to the para-position of the ferrocenyl group. Introducing the ferrocenyl group into natural compounds may be a useful strategy for increasing the antioxidative effectiveness.

Protection of Clitoria ternatea flower petal extract against free radical-induced hemolysis and oxidative damage in canine erythrocytes

The present study assessed the antioxidant activity and protective ability of Clitoria ternatea flower petal extract (CTE) against in vitro 2,2′-azobis-2-methyl-propanimidamide dihydrochloride (AAPH)-induced hemolysis and oxidative damage of canine erythrocytes. From the phytochemical analysis, CTE contained phenolic compounds, flavonoids, and anthocyanins. In addition, CTE showed antioxidant activity as measured by oxygen radical absorbance capacity (ORAC) method and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. CTE (400 µg/ml) remarkably protected erythrocytes against AAPH-induced hemolysis at 4 h of incubation. Moreover, CTE (400 µg/ml) reduced membrane lipid peroxidation and protein carbonyl group formation and prevented the reduction of glutathione concentration in AAPH-induced oxidation of erythrocytes. The AAPH-induced morphological alteration of erythrocytes from a smooth discoid to an echinocytic form was effectively protected by CTE. The present results contribute important insights that CTE may have the potential to act as a natural antioxidant to prevent free radical-induced hemolysis, protein oxidation and lipid peroxidation in erythrocytes.

Design and synthesis of coumarin-3-acylamino derivatives to scavenge radicals and to protect DNA

In this study, a series of coumarin-3-acylamino derivatives containing phenethylamine moiety or tyramine moiety were synthesized and their antioxidant activities were evaluated by Cu2+/glutathione(GSH)-, OH- and 2,2′-azobis(2-amidinopropane hydrochloride)(AAPH)-induced oxidation of DNA. It was found that both hydroxyl and ortho-methoxy groups at A ring, hydroxyl group at B ring and peptide bond can enhance the abilities of coumarin-3-acylamino derivatives to protect DNA against OH- and AAPH-induced oxidation. Moreover, these coumarin-3-acylamino derivatives were employed to scavenge 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS+). We found that tyramine moiety, hydroxyl and ortho-methoxy are the key groups to enhance the activities of antioxidants to quench ABTS+. Therefore, tyramine linked with coumarin-3-carboxyl acid which containing hydroxyl and ortho-methoxy exhibited powerful antioxidant abilities.

Coumestan inhibits radical-induced oxidation of DNA: is hydroxyl a necessary functional group?

Coumestan is a natural tetracycle with a C═C bond shared by a coumarin moiety and a benzofuran moiety. In addition to the function of the hydroxyl group on the antioxidant activity of coumestan, it is worth exploring the influence of the oxygen-abundant scaffold on the antioxidant activity as well. In this work, seven coumestans containing electron-withdrawing and electron-donating groups were synthesized to evaluate the abilities to trap 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS(•+)), 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH), and galvinoxyl radical, respectively, and to inhibit the oxidations of DNA mediated by (•)OH, Cu(2+)/glutathione (GSH), and 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH), respectively. It was found that all of the coumestans used herein can quench the aforementioned radicals and can inhibit (•)OH-, Cu(2+)/GSH-, and AAPH-induced oxidations of DNA. In particular, substituent-free coumestan exhibits higher ability to quench DPPH and to inhibit AAPH-induced oxidation of DNA than Trolox. In addition, nonsubstituted coumestan shows a similar ability to inhibit (•)OH- and Cu(2+)/GSH-induced oxidations of DNA relative to that of Trolox. The antioxidant effectiveness of the coumestan can be attributed to the lactone in the coumarin moiety and, therefore, a hydroxyl group may not be a necessary functional group for coumestan to be an antioxidant.

Antioxidant action of SMe1EC2, the low-basicity derivative of the pyridoindole stobadine, in cell free chemical models and at cellular level.

The aim of the study was to evaluate the antioxidant action of SMe1EC2, the structural analogue of the hexahydropyridoindole antioxidant stobadine. The antiradical activity of SMe1EC2 was found to be higher when compared to stobadine, as determined both in cell-free model systems of AAPH-induced oxidation of dihydrorhodamine 123 and 2′,7′-dichloro-dihydrofluorescein diacetate, and in the cellular system of stimulated macrophages RAW264.7. Analysis of proliferation of HUVEC and HUVEC-ST cells revealed absence of cytotoxic effect of SMe1EC2 at concentrations below 100 µM. The antioxidant activity of SMe1EC2, superior to the parent drug stobadine, is accounted for by both the higher intrinsic free radical scavenging action and by the better bioavailability of the low-basicity SMe1EC2 relative to the high-basicity stobadine.