RESUMEN
Nonsymbiotic phytoglobins (nsHbs) are a diverse superfamily of hemoproteins grouped into three different classes (1, 2, and 3) based on their sequences. Class 1 Hb are expressed under hypoxia, osmotic stress, and/or nitric oxide exposure, while class 2 Hb are induced by cold stress and cytokinins. Both are mainly six-coordinated. The deoxygenated forms of the class 1 and 2 nsHbs from A. thaliana (AtHb1 and AtHb2) are able to reduce nitrite to nitric oxide via a mechanism analogous to other known globins. NsHbs provide a viable pH-dependent pathway for NO generation during severe hypoxia via nitrite reductase-like activity with higher rate constants compared to mammalian globins. These high kinetic parameters, along with the relatively high concentrations of nitrite present during hypoxia, suggest that plant hemoglobins could indeed serve as anaerobic nitrite reductases in vivo. The third class of nsHb, also known as truncated hemoglobins, have a compact 2/2 structure and are pentacoordinated, and their exact physiological role remains mostly unknown. To date, no reports are available on the nitrite reductase activity of the truncated AtHb3. In the present work, three representative nsHbs of the plant model Arabidopsis thaliana are presented, and their nitrite reductase-like activity and involvement in nitrosative stress is discussed. The reaction kinetics and mechanism of nitrite reduction by nsHbs (deoxy and oxy form) at different pHs were studied by means of UV-Vis spectrophotometry, along with EPR spectroscopy. The reduction of nitrite requires an electron supply, and it is favored in acidic conditions. This reaction is critically affected by molecular oxygen, since oxyAtHb will catalyze nitric oxide deoxygenation. The process displays unique autocatalytic kinetics with metAtHb and nitrate as end-products for AtHb1 and AtHb2 but not for the truncated one, in contrast with mammalian globins.
Asunto(s)
Arabidopsis , Nitritos , Animales , Nitritos/química , Óxido Nítrico/metabolismo , Hemoglobinas/química , Nitrito Reductasas/química , Hipoxia , Arabidopsis/metabolismo , Oxidación-Reducción , Mamíferos/metabolismoRESUMEN
In this research, we investigated the structural and biological properties of phosphate glasses (PGs) after the addition of V2O5. A xV2O5â(100 − x)[CaF2â3P2O5âCaO] glass system with 0 ≤ x ≤ 16 mol% was synthesized via a conventional melt-quenching technique. Several analysis techniques (dissolution tests, pH, SEM-EDS, FT-IR, and EPR) were used to obtain new experimental data regarding the structural behavior of the system. In vitro tests were conducted to assess the antitumor character of V2O5-doped glass (x = 16 mol%) compared to the matrix (x = 0 mol%) and control (CTRL-) using several tumoral cell lines (A375, A2780, and Caco-2). The characterization of PGs showed an overall dissolution rate of over 90% for all vitreous samples (M and V1−V7) and the high reactivity of this system. EPR revealed a well-resolved hyperfine structure (hfs) typical of vanadyl ions in a C4v symmetry. FT-IR spectra showed the presence of all structural units expected for P2O5, as well as very clear depolymerization of the vitreous network induced by V2O5. The MTT assay indicated that the viability of tumor cells treated with V7-glass extract was reduced to 50% when the highest concentration was used (10 µg/mL) compared to the matrix treatment (which showed no cytotoxic effect at any concentration). Moreover, the matrix treatment (without V2O5) provided an optimal environment for tumor cell attachment and proliferation. In conclusion, the two types of treatment investigated herein were proven to be very different from a statistical point of view (p < 0.01), and the in vitro studies clearly underline the cytotoxic potential of vanadium ions from phosphate glass (V7) as an antitumor agent.
Asunto(s)
Neoplasias Ováricas , Vanadio , Femenino , Humanos , Vanadio/farmacología , Espectroscopía Infrarroja por Transformada de Fourier , Línea Celular Tumoral , Células CACO-2 , Vanadatos , Fosfatos , Vidrio/química , IonesRESUMEN
Copper is one of the most used therapeutic metallic elements in biomedicine, ranging from antibacterial approaches to developing new complexes in cancer therapy. In the present investigation, we developed a novel xCuOâ(100 - x) [CaF2â3P2O5âCaO] glass system with 0 ≤ x ≤ 16 mol% in order to determine the influence of doping on the composition structure of glasses. The samples were characterized by dissolution tests, pH measurements, Fourier-transform infrared spectroscopy (FT-IR), electron paramagnetic resonance (EPR), Scanning Electron Microscopy with energy dispersive spectroscopy (SEM-EDX) and afterward, their antitumor character was assessed. The glasses were mostly soluble in the aqueous medium, their dissolution rate being directly proportional to the increase in pH and the level of doping up to x = 8 mol%. FT-IR spectra of glass samples show the presence of all structural units characteristic to P2O5 in different rates and directly depending on the depolymerization process. SEM-EDX results revealed the presence of an amorphous glass structure composed of P, O, Ca, and Cu elements. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay showed strong cytotoxicity for tumoral cells A375 even in low concentrations for Cu-treatment. In contrast, the copper-free matrix (without Cu) determined a proliferative effect of over 70% viability for all concentrations used.
RESUMEN
Yellow laccases lack the typical blue type 1 Cu absorption band around 600 nm; however, multi-copper oxidases with laccase properties have been reported. We provide the first evidence that the yellow laccase isolated from Sclerotinia sclerotiorum is obtained from a blue form by covalent, but nevertheless reversible modification with a phenolic product. After separating the phenolics from the extracellular medium, a typical blue laccase is obtained. With ABTS as model substrate for this blue enzyme, a non-natural purple adduct is formed with a spectrum nearly identical to that of the 1:1 adduct of an ABTS radical and Tyr. This modification significantly increases the stability and substrate affinity of the enzyme, not by acting primarily as bound mediator, but by structural changes that also alters the type 1 Cu site. The HPLC-MS analyses of the ABTS adduct trypsin digests revealed a distinct tyrosine within a unique loop as site involved in the modification of the blue laccase form. Thus, S. sclerotiorum yellow laccase seems to be an intrinsically blue multi-copper oxidase that boosts its activity and stability with a radical-forming aromatic substrate. This particular case could, at least in part, explain the enigma of the yellow laccases.
Asunto(s)
Ascomicetos/enzimología , Lacasa/metabolismo , Tirosina/metabolismo , Biocatálisis , Color , Concentración de Iones de Hidrógeno , Fenoles/metabolismo , Unión ProteicaRESUMEN
We have previously proposed the annelid-derived protein, hemerythrin, as a viable replacement for hemoglobin in the synthesis of semi-synthetic oxygen carriers ("blood substitutes"). Here, we report the first in vivo tests for potential hemerythrin-based oxygen carriers (HrBOC), using a battery of experiments involving Wistar rats and previously tested on a series of hemoglobin-based oxygen carrier candidates (HBOC). At the concentrations tested, hemerythrin appears to behave similarly to hemoglobin - including, importantly, immunological effects. The antioxidant strategies based on albumin as well as based on rubrerythrin appear to offer observable physiological advantages.
Asunto(s)
Hemeritrina/metabolismo , Hemoglobinas/metabolismo , Oxígeno/metabolismo , Animales , Coagulación Sanguínea , Catalasa/metabolismo , Glucosa/análisis , Iones , Hierro/metabolismo , Masculino , Estrés Oxidativo , Ratas Wistar , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
The present study reports findings regarding the contrast between H2S interaction with bovine hemoglobin (Hb) and horse heart myoglobin (Mb), in terms of binding and dissociation kinetics, affinities, and mechanism. At pH9.5, oxidation of ferric-sulfide adducts in presence of no free sulfide, using hexachloroiridate as oxidant is examined using stopped-flow UV-vis, EPR, vibrational spectroscopy and mass spectrometry. Oxidation of the ferric-sulfide adduct in such conditions occurs with a putative unstable Fe(IV)-sulfide adduct as intermediate that finally leads to a paramagnetic ferric species with distinct EPR features. As detected by MS spectrometry, this final species appears to be a truncated form of globin at a distinct Tyr. In case of Hb, only ß-chain is truncated at Tyr144.
Asunto(s)
Hemoglobinas/química , Sulfuro de Hidrógeno/química , Hierro/química , Mioglobina/química , Animales , Bovinos , Caballos , Concentración de Iones de Hidrógeno , Cinética , Miocardio/química , Oxidación-ReducciónRESUMEN
Stopped-flow UV-vis data allow for the first time direct spectroscopic detection of a ferryl species during the reaction of met myoglobin (Mb) with chlorite, analogous to what is observed in the reaction with peroxides. Ferryl is also observed in the reaction of oxy Mb+chlorite. A pathway involving Fe-O-O-ClO2 is explored by analogy with the Fe-O-O-NO and Fe-O-O-NO2 previously proposed as intermediates in the reactions of oxy globins with nitric oxide and nitrite, respectively. However, Fe-O-O-ClO2 is not detectable in these stopped-flow experiments and is in fact, unlike its nitrogenous congeners, predicted by density functional theory (DFT) to be impossible for a heme complex. Deoxy Mb reacts with chlorite faster than met - suggesting that, unlike with hydrogen peroxide (with which deoxy Mb reacts slower than met), binding of chlorite to the heme is not a rate-determining step (hence, most likely, an outer-sphere electron transfer mechanism); to correlate this, a Fe-O-Cl-O adduct was not observed experimentally for the met or for the deoxy reactions - even though prior DFT calculations suggest it to be feasible and detectable.
Asunto(s)
Cloruros/química , Mioglobina/química , Nitritos/química , Peróxidos/química , Teoría Cuántica , Hemo/química , Modelos QuímicosRESUMEN
A series of hemoglobin-based oxygen carrier candidates (HBOC), previously noted for their differences in prooxidative and physiological reactivity, were compared in terms of the negative effects displayed upon injection in Wistar rats. At the concentrations tested, antioxidant strategies based on albumin as well as based on rubrerythrin appear to offer observable physiological advantages.
Asunto(s)
Sustitutos Sanguíneos/farmacología , Hemoglobinas/farmacología , Especies Reactivas de Oxígeno/metabolismo , Animales , Coagulación Sanguínea/efectos de los fármacos , Glucemia/metabolismo , Sustitutos Sanguíneos/efectos adversos , Bovinos , Pruebas Hematológicas , Hemoglobinas/efectos adversos , Hierro/metabolismo , Masculino , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Wistar , Transferrina/metabolismoRESUMEN
Two new protocols for exploring antioxidant-related chemical composition and reactivity are described: one based on a chronometric variation of a haemoglobin ascorbate peroxidase assay and one based on cytochrome c-induced oxidation of lecithin liposomes. Detailed accounts are given on their design, application, critical correlations with established methods and mechanisms. These assays are proposed to be physiologically relevant and bring new information regarding a real sample, both qualitative and quantitative. The well-known assays used for evaluation of antioxidant (re)activity are revisited and compared with these new methods. Extracts of the Hedera helix L. are examined as test case, with focus on seasonal variation and on leaf, fruit and flower with respect to chromatographic, spectroscopic and reactivity properties. According to the set of assays performed, winter are the most antioxidant, followed by summer leaves, and then by flowers and fruits.
Asunto(s)
Antioxidantes/farmacología , Liposomas/química , Peroxidasas/metabolismo , Animales , Antioxidantes/química , Bioensayo/métodos , Citocromos c/farmacología , Flores/química , Frutas/química , Hemoglobinas , Humanos , Oxidación-Reducción/efectos de los fármacos , Extractos Vegetales/química , Hojas de la Planta/química , Estaciones del AñoRESUMEN
Two methods for the measurement of antioxidant capacity are described: one based on a chronometric variation of a hemoglobin ascorbate peroxidase assay and the other based on electron paramagnetic resonance (EPR) spectra collected upon alkaline treatment of ethanolic samples. The involved chemical mechanisms are discussed, alongside the most important benefits and shortcomings; the assays offer new qualitative and quantitative information on samples of biological as well as synthetic origin.
Asunto(s)
Antioxidantes/metabolismo , Espectroscopía de Resonancia por Spin del Electrón/métodos , Radicales Libres/metabolismo , Hemoglobinas/metabolismo , Animales , Ascorbato Peroxidasas/metabolismo , Ácido Ascórbico/metabolismo , Bovinos , Oxidación-Reducción , Factores de TiempoRESUMEN
The reaction between nitrite and the oxy forms of globins has complex autocatalytic kinetics with several branching steps and evolves through chain reactions mediated by reactive species (including radicals) such as hydrogen peroxide, ferryl and nitrogen dioxide, starting with a lag phase, after which it proceeds onto an autocatalytic phase. Reported here are UV-Vis spectra collected upon stopped-flow mixing of myoglobin with a supraphysiological excess of nitrite. The best fit to the experimental data follows an A â B â C reaction scheme involving the formation of a short-lived intermediate identified as ferryl. This is consistent with a mechanism where nitrite binds to oxy myoglobin to generate an undetectable ferrous-peroxynitrate intermediate, whose decay leads to nitrate and ferryl. The ferryl is then reduced to met by the excess nitrite. DFT calculations reveal an essentially barrierless reaction between nitrite and the oxy heme, with a notable outer-sphere component; the resulting metastable ferrous-peroxynitrate adduct is found to feature a very low barrier towards nitrate liberation, with ferryl as a final product-in good agreement with experiment.
Asunto(s)
Hierro/metabolismo , Mioglobina/metabolismo , Nitritos/metabolismo , Ácido Peroxinitroso/metabolismo , Cachalote/metabolismo , Animales , Hierro/química , Modelos Moleculares , Mioglobina/química , Nitritos/química , Oxidación-Reducción , Ácido Peroxinitroso/químicaRESUMEN
This study was designed to examine the in vitro antioxidant and antimicrobial activities and to characterize the polyphenolic composition of the ethanolic extracts of Hyssopus officinalis, Ocimum basilicum and Teucrium chamaedrys. Qualitative and quantitative analysis of the major phenolic compounds were conducted using high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS). The total polyphenols, caffeic acid derivatives and flavonoids content was spectrophotometrically determined. The phenolic profile showed the presence of phenolic acid derivatives (caftaric, gentisic, caffeic, p-coumaric, chlorogenic and ferulic acids), flavonoid glycosides (rutin, isoquercitrin and quercitrin) and free flavonoid aglycons (luteolin, quercetin), in different concentrations. DPPH radical scavenging assay, Trolox equivalent antioxidant capacity (TEAC) method, hemoglobin ascorbate peroxidase activity inhibition (HAPX) assay, and electron paramagnetic resonance (EPR) radicals detection were employed, revealing several aspects of the antioxidant activities of these species. The antimicrobial tests were performed using the disk diffusion assay. These extracts contained a large amount of the polyphenolic compounds (77.72, 175.57, and 243.65 mg/g, respectively), and they showed a good antioxidant activity, as witnessed by a number of methods. T. chamaedrys had a high antimicrobial activity. Besides their antioxidant activity, the antimicrobial effect of these extracts confirms the biological activities of these herbal medicinal products.
Asunto(s)
Antiinfecciosos/farmacología , Antioxidantes/aislamiento & purificación , Bacterias/efectos de los fármacos , Fenoles/aislamiento & purificación , Antiinfecciosos/aislamiento & purificación , Antioxidantes/química , Antioxidantes/farmacología , Lamiaceae/química , Ocimum basilicum/química , Fenoles/farmacología , Extractos Vegetales , Teucrium/químicaRESUMEN
The nonheme peroxidase, rubrerythrin, shows the ability to reduce hydrogen peroxide to water without involving strongly oxidizing and free-radical-creating powerful oxidants such as compounds I and II [formally Fe(IV)] formed in peroxidases and catalases. Rubrerythrin could, therefore, be a useful ingredient in protein-based artificial oxygen carriers. Here, we report that the oxygen-carrying proteins, hemoglobin (Hb) and hemerythrin (Hr), can each be copolymerized with rubrerythrin using glutaraldehyde yielding high molecular weight species. These copolymers show additional peroxidase activity compared to Hb-only and Hr-only polymers, respectively and also generate lower levels of free radicals in reactions that involve hydrogen peroxide. Tests on human umbilical vein endothelial cells (HUVEC) reveal slightly better performance of the Rbr copolymers compared to controls, as measured at 24 h, but not at later times.
Asunto(s)
Sustitutos Sanguíneos/metabolismo , Hemeritrina/metabolismo , Hemoglobinas/metabolismo , Oxígeno/metabolismo , Peroxidasa/metabolismo , Sustitutos Sanguíneos/química , Hemeritrina/química , Hemoglobinas/química , Células Endoteliales de la Vena Umbilical Humana/química , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Modelos Moleculares , Conformación Molecular , Oxidación-Reducción , Oxígeno/química , Peroxidasa/químicaRESUMEN
A transient species may be detected with UV-vis and EPR spectroscopy during turnover of a laccase with quercetin; this species is assigned as a quercetin-derived radical, based on EPR spectra as well the observed UV-vis similarities (a 540nm centred band) with previously reported data. The rates of formation and decay of this species correlate well (r=0.9946) with the pro-oxidant reactivity manifested by flavonoids in the presence of laccase. An assay for the pro-oxidant reactivity of natural products is hence proposed based on the results reported here; its application is demonstrated for a series of pure compounds as well as for several propolis extracts. This assay has the advantages of using a biologically relevant process (haemoglobin oxidation), and not requiring the addition of oxidising agents such as peroxide or superoxide. Correlations, or the lack thereof, between the pro-oxidant parameters and the redox potentials, antioxidant capacities and lipophilicities, were analysed. The laccase employed in our study does display reactivity-related similarities to a range of other proteins, including human plasma ceruloplasmin.
Asunto(s)
Ascomicetos/enzimología , Proteínas Fúngicas/química , Lacasa/química , Oxidantes/química , Fenoles/química , Espectroscopía de Resonancia por Spin del Electrón , Flavonoides/química , Cinética , Oxidación-Reducción , Quercetina/químicaRESUMEN
The effects of gamma radiation are investigated by studying plant germination, growth and development, and biochemical characteristics of maize. Maize dry seeds are exposed to a gamma source at doses ranging from 0.1 to 1 kGy. Our results show that the germination potential, expressed through the final germination percentage and the germination index, as well as the physiological parameters of maize seedlings (root and shoot lengths) decreased by increasing the irradiation dose. Moreover, plants derived from seeds exposed at higher doses (≤0.5 kGy) did not survive more than 10 days. Biochemical differences based on photosynthetic pigment (chlorophyll a, chlorophyll b, carotenoids) content revealed an inversely proportional relationship to doses of exposure. Furthermore, the concentration of chlorophyll a was higher than chlorophyll b in both irradiated and non-irradiated seedlings. Electron spin resonance spectroscopy used to evaluate the amount of free radicals induced by gamma ray treatment demonstrates that the relative concentration of radiation-induced free radicals depends linearly on the absorbed doses.
Asunto(s)
Radicales Libres/metabolismo , Rayos gamma , Germinación/efectos de la radiación , Pigmentos Biológicos/metabolismo , Semillas/crecimiento & desarrollo , Semillas/efectos de la radiación , Zea mays/efectos de la radiación , Espectroscopía de Resonancia por Spin del Electrón , Fotosíntesis/efectos de la radiación , Tolerancia a Radiación , Zea mays/crecimiento & desarrollo , Zea mays/metabolismoRESUMEN
We aimed at investigating the antioxidant, antiinflamatory, antiapoptotic and antigenotoxic effects of a Romanian Propolis (RP) extract in two concentrations (RP1 3 mg, respectively RP2 1.5 mg polyphenols/cm(2)), topically administered, either prior to or after UVB exposure, in a Swiss mouse model. Our results showed that both concentrations of RP extract, independent of the time of administration, significantly attenuated the malondialdehyde (MDA) formation and restored glutathione peroxidase (GPx) activity. However, the 8-hydroxy-2-deoxy-guanosine (8-oxo-dG), nitric oxide (NO) and glutathione (GSH) levels were not influenced by UVB exposure and RP treatment. Interleukin (IL)-6 levels were significantly decreased by RP treatment, both before and after UVB-exposure. RP2 extract, in both regimens, significantly reduced the epidermal hyperplasia and dermal inflammation, whereas RP1 pre-treatment diminished only the dermal inflammation. The effect of our RP extract in terms of reduction of sunburn cell formation and of activated caspase-3 and TUNEL-positive cells was observed in both subsets of the experiment, RP2 having a slightly better protective effect as compared to RP1. The antigenotoxic effect of RP was demonstrated by significantly reduced cyclobutane pyrimidine dimers (CPDs) formation. Our results suggest that RP extract might be a potential chemopreventive candidate by modulation of multiple UVB-induced signaling pathways in skin.
Asunto(s)
Própolis/química , Própolis/farmacología , Protectores contra Radiación/farmacología , Piel/efectos de los fármacos , Piel/efectos de la radiación , 8-Hidroxi-2'-Desoxicoguanosina , Administración Tópica , Animales , Antioxidantes/química , Antioxidantes/farmacología , Caspasa 3/metabolismo , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Femenino , Glutatión/metabolismo , Hiperplasia/tratamiento farmacológico , Interleucina-6/metabolismo , Ratones , Óxido Nítrico/metabolismo , Estrés Oxidativo/efectos de los fármacos , Polifenoles/análisis , Polifenoles/farmacología , Dímeros de Pirimidina/metabolismo , Radiodermatitis/tratamiento farmacológico , Radiodermatitis/patología , Rumanía , Piel/metabolismo , Piel/patología , Rayos Ultravioleta/efectos adversosRESUMEN
The aim of this work was to study the chemical composition, antioxidant and antibacterial properties of Achillea distans Waldst. et Kit. subsp. distans and Achillea distans Waldst. et Kit. subsp. alpina Rochel, from the Rodna Mountains (Romania). The identification and quantification of major phenolic compounds was performed by a HPLC-MS method. The total polyphenolic and flavonoid content was determined spectrophotometrically. The antioxidant activity was evaluated using the DPPH bleaching method, trolox equivalent antioxidant capacity assay (TEAC), hemoglobin ascorbate peroxidase activity inhibition (HAPX) assay, and an Electron Paramagnetic Resonance (EPR) spectroscopy method. A data indicated that A. distans subsp. alpina extract has more antioxidant activity than A. distans subsp. distans extract. Luteolin, apigenin, quercetin, caffeic and chlorogenic acids were present in the two extracts of A. distans, but in different amounts. Three flavonoids were detected only in A. distans subsp. alpina. The polyphenol-richer A. distans subsp. alpina extract showed a higher antioxidant activity than A. distans subsp. distans extract. A. distans subsp. distans extract showed inhibitory activity for Gram-positive bacteria, as evaluated with four species. The quantitative and qualitative differences between the two subspecies of Achillea distans could be used as a potential taxonomic marker in order to distinguish the species.
Asunto(s)
Achillea/química , Bacterias Grampositivas/efectos de los fármacos , Extractos Vegetales/farmacología , Polifenoles/aislamiento & purificación , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Cromatografía Líquida de Alta Presión , Espectrometría de Masas , Extractos Vegetales/química , Polifenoles/química , Polifenoles/farmacología , Rumanía , Especificidad de la EspecieRESUMEN
Traditional remedies have a long-standing history in Cameroon and continue to provide useful and applicable tools for treating ailments. Here, the anticancer, antimicrobial and antioxidant activities of ten antioxidant-rich Cameroonian medicinal plants and of some of their isolated compounds are evaluated.The plant extracts were prepared by maceration in organic solvents. Fractionation of plant extract was performed by column chromatography and the structures of isolated compounds (emodin, 3-geranyloxyemodin, 2-geranylemodin) were confirmed spectroscopically. The antioxidant activity (AOA) was determined using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) bleaching method, the trolox equivalent antioxidant capacity (TEAC), and the hemoglobin ascorbate peroxidase activity inhibition (HAPX) assays. The anticancer activity was evaluated against A431 squamous epidermal carcinoma, WM35 melanoma, A2780 ovary carcinoma and cisplatin-resistant A2780cis cells, using a direct colorimetric assay. The total phenolic content in the extracts was determined spectrophotometrically by the Folin-Ciocalteu method. Rumex abyssinicus showed the best AOA among the three assays employed. The AOA of emodin was significantly higher than that of 3-geranyloxyemodin and 2-geranylemodin for both TEAC and HAPX methods. The lowest IC(50) values (i.e., highest cytotoxicity) were found for the extracts of Vismia laurentii, Psorospermum febrifugum, Pentadesma butyracea and Ficus asperifolia. The Ficus asperifolia and Psorospermum febrifugum extracts are selective against A2780cis ovary cells, a cell line which is resistant to the standard anticancer drug cisplatin. Emodin is more toxic compared to the whole extract, 3-geranyloxyemodin and 2-geranylemodin. Its selectivity against the platinum-resistant A2780cis cell line is highest. All of the extracts display antimicrobial activity, in some cases comparable to that of gentamycin.
Asunto(s)
Antiinfecciosos/farmacología , Antineoplásicos/farmacología , Antioxidantes/farmacología , Extractos Vegetales/química , Plantas Medicinales/química , Antiinfecciosos/química , Antiinfecciosos/toxicidad , Antineoplásicos/química , Antineoplásicos/toxicidad , Antioxidantes/química , Antioxidantes/toxicidad , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Análisis por Conglomerados , Relación Dosis-Respuesta a Droga , Humanos , Concentración 50 Inhibidora , Pruebas de Sensibilidad Microbiana , Fenol/química , Fenol/farmacología , Componentes Aéreos de las Plantas/química , Análisis EspectralRESUMEN
We have previously reported that derivatization of hemoglobin with periodate-modified sugar derivatives such as oxidized adenosine triphosphate (oATP) leads to an increase in prooxidant reactivity at the heme. Here, we report that copolymerization of hemoglobin with serum albumin alleviates this problem completely, to the extent where the copolymer even has a slightly lower autooxidation rate compared to native hemoglobin. A similar, although not as potent, effect is obtained when hemoglobin is derivatized with oATP in the presence of small-molecule antioxidants instead of albumin.
Asunto(s)
Antioxidantes , Sustitutos Sanguíneos/metabolismo , Hemo/metabolismo , Hemoglobinas/metabolismo , Albúmina Sérica , Adenosina Trifosfato/análogos & derivados , Adenosina Trifosfato/química , Adenosina Trifosfato/metabolismo , Antioxidantes/química , Antioxidantes/metabolismo , Sustitutos Sanguíneos/química , Borohidruros/química , Reactivos de Enlaces Cruzados/química , Espectroscopía de Resonancia por Spin del Electrón , Electroforesis en Gel de Poliacrilamida , Hemo/química , Hemoglobinas/química , Humanos , Oxidación-Reducción , Polimerizacion , Especies Reactivas de Oxígeno/metabolismo , Albúmina Sérica/química , Albúmina Sérica/metabolismo , Análisis EspectralRESUMEN
Periodate modification of the sugar moiety in sugars, including adenosine triphosphate (ATP), has previously been employed in order to prepare dialdehyde-type reagents, which were then utilized in crosslinking reactions on haemoglobin, yielding polymerized material with useful dioxygen-binding properties and hence proposed as possible artificial oxygen carriers ('blood substitutes'). Here, the periodate protocol is shown to be applicable to a wider range of oxygen-containing compounds, illustrated by starch and polyethylene glycol. Derivatization protocols are described for haemoglobin with such periodate-treated crosslinking agents, and the dioxygen-binding properties and redox reactivities are investigated for the derivatized haemoglobins, with emphasis on pro-oxidative properties. There is a general tendency of the derivatization to result in higher autooxidation rates. The peroxide reactivity of the met (ferric) form is also affected by derivatization, as witnessed, among others, by varying yields of ferryl [Fe (IV)-oxo] and free radical generated. In cell, culture tests (human umbilical vein epithelial cells, HUVEC), the derivatization protocols show no toxic effect.