RESUMO
BACKGROUND: Psoriasis is a chronic, inflammatory skin condition associated with a high frequency of cardiovascular events. Modifications of plasma lipids, and an increase in the levels of biochemical markers of inflammation and lipid peroxidation have been reported in subjects with psoriasis, suggesting a relationship between psoriasis, inflammation and oxidative damage. OBJECTIVES: To investigate whether modulation of inflammatory activity by tumour necrosis factor-α inhibitors in patients with psoriasis is associated with modification of lipid profiles, oxidative stress and paraoxonase (PON)1 activity. METHODS: The levels of plasma lipids and lipoprotein(a), and the levels of the markers of inflammation and lipid peroxidation were evaluated in subjects with psoriasis (n=23) before and after 24 weeks of treatment with etanercept. In the same subjects plasma total antioxidant capacity and the activity of PON1, an antioxidant and anti-inflammatory enzyme associated with the high-density lipoproteins (HDLs), were investigated. RESULTS: The results showed that clinical improvement in patients with psoriasis treated with etanercept is associated with a reduction in the levels of inflammatory markers [C-reactive protein (CRP)] and lipid peroxidation, and also with increased antioxidant capacity in the serum of patients with psoriasis. These modifications are associated with a significant increase in the activity of PON1. A significant increase in the PON1/CRP ratio has also been observed in patients with psoriasis after treatment. The significant inverse correlation between CRP and PON1 activity suggests a relationship between PON1 activity and inflammation. CONCLUSIONS: Treatment with etanercept is associated with a reduction in lipid peroxidation and an improvement in HDL antioxidant and anti-inflammatory properties.
Assuntos
Imunoglobulina G/uso terapêutico , Imunossupressores/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Psoríase/tratamento farmacológico , Receptores do Fator de Necrose Tumoral/uso terapêutico , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adulto , Arildialquilfosfatase/metabolismo , Biomarcadores/metabolismo , Proteína C-Reativa/metabolismo , Etanercepte , Feminino , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Lipídeos/sangue , Lipoproteína(a)/sangue , Lipoproteínas HDL/metabolismo , Masculino , Pessoa de Meia-Idade , Oxirredução/efeitos dos fármacos , Psoríase/sangue , Índice de Gravidade de Doença , Fatores de TempoRESUMO
BACKGROUND: Psoriasis is a chronic, inflammatory skin disease associated with abnormal plasma lipid metabolism and with a high frequency of cardiovascular events. Modifications of plasma lipids and an increase in the levels of biochemical markers of lipid peroxidation have been reported in subjects with psoriasis, suggesting a relationship between psoriasis, lipoproteins and oxidative damage. OBJECTIVES: To investigate further the relationship between lipoproteins and oxidative stress in psoriasis. METHOD: The levels of plasma lipids, lipoprotein(a) [Lp(a)] and markers of lipid peroxidation were evaluated in subjects with psoriasis (n=23) and in controls (n=25). In the same subjects, the activity of paraoxonase-1 (PON1), an antioxidant and an anti-inflammatory enzyme associated with high-density lipoproteins, was investigated. RESULTS: The results showed higher levels of Lp(a) in the serum of patients with psoriasis compared with controls (P<0·001). Higher levels of lipid hydroperoxides (P<0·001) and lower PON1 activity were observed in the serum of patients compared with healthy subjects, confirming that psoriasis is associated with oxidative stress. The imbalance between oxidative stress and antioxidant enzymes, and the increase of Lp(a) serum levels was related to the extent and severity of psoriasis. Finally, our results demonstrated that Lp(a) levels were positively correlated with markers of lipid peroxidation and negatively related to PON1 activity, suggesting that subjects with higher levels of Lp(a) are more exposed to oxidative damage. CONCLUSIONS: Our results provide further evidence that oxidative stress and impairment of the antioxidant system in the plasma of patients may play a role in pathogenesis and progression of psoriasis and related complications.
Assuntos
Arildialquilfosfatase/fisiologia , Peroxidação de Lipídeos/fisiologia , Lipoproteína(a)/metabolismo , Estresse Oxidativo/fisiologia , Psoríase/enzimologia , Adulto , Arildialquilfosfatase/metabolismo , Biomarcadores/metabolismo , Estudos de Casos e Controles , Doença Crônica , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
AIMS: Aim of the paper was to summarize the literature about the effect of dietary lipids on activity of paraoxonase-1 (PON1), a multifunctional enzyme associated with high density lipoprotein (HDL). PON1 exerts a protective effect against oxidative damage of cells and lipoproteins and modulates the susceptibility of HDL and LDL to atherogenic modifications such as homocysteinylation. DATA SYNTHESIS: The present review shows evidence that the amount and the composition of dietary lipids are key factors in the modulation of PON1. The effect of dietary lipids is also modulated by PON1 polymorphisms. The molecular mechanisms involved include an effect on PON1 hepatic synthesis or secretion and/or modification of PON1 interactions with HDL. Changes of PON1 activity could also be related to dietary intake of oxidized lipids that behave as PON1 inhibitors. CONCLUSION: Dietary fatty acids by the modulation of PON1 gene expression and activity could constitute an useful approach for the prevention of human diseases associated with oxidative damage.
Assuntos
Arildialquilfosfatase/metabolismo , Gorduras na Dieta/administração & dosagem , Regulação Enzimológica da Expressão Gênica , Animais , Arildialquilfosfatase/genética , Humanos , Lipoproteínas HDL/metabolismo , Lipoproteínas LDL/metabolismo , Nutrigenômica , Polimorfismo GenéticoAssuntos
Aorta/citologia , Diabetes Mellitus Tipo 1/metabolismo , Células Endoteliais/metabolismo , Homocisteína/metabolismo , Lipoproteínas LDL/metabolismo , Adulto , Estudos de Casos e Controles , Células Cultivadas , Diabetes Mellitus Tipo 1/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo/efeitos dos fármacos , Fatores de RiscoRESUMO
Oxidative damage induced by free radicals and reactive oxygen species (ROS) have been suggested to play an important role in the development of autoimmune diseases such as multiple sclerosis (MS) disease and it has been hypothesised that oxidative injury could mediate demyelination and axonal injury in MS subjects. In our study, we compared intracellular oxidative activity and the respiratory burst activity in MS patients (n=20) and healthy controls (n=15) using leukocytes as cellular model. At this purpose, intracellular ROS levels were evaluated by fluorometric assay using the 2'-7'-dichlorodihydrofluorescin diacetate probe (H(2)DCFDA) in untreated or in leukocytes stimulated with phorbol-12-myristate-13-acetate (PMA). Our results demonstrate that the intracellular spontaneous ROS production in leukocytes from MS patients was higher with respect to cells from control subjects (p<0.001). PMA addition induced a higher formation of ROS both in leukocytes from MS patients and controls (p<0.001). The PMA-induced production of ROS was significantly higher in leukocytes from MS with respect to controls (p<0.001). Significant positive correlations were established between intracellular spontaneous or PMA-induced production of ROS in leukocytes isolated from MS patients and the clinical parameters used to evaluate disease disability such as expanded disability status scale (EDSS), brain lesions evaluated by MRI and visual evoked potential (VEP) (p<0.001). In conclusion, our results demonstrate higher levels of intracellular ROS in untreated or in PMA-treated leukocytes isolated from MS patients with respect to healthy subjects confirming the role of oxidative stress in multiple sclerosis.
Assuntos
Leucócitos/metabolismo , Esclerose Múltipla/sangue , Estresse Oxidativo , Adulto , Estudos de Casos e Controles , Potenciais Evocados Visuais , Feminino , Fluoresceínas/química , Corantes Fluorescentes/química , Humanos , Leucócitos/efeitos dos fármacos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/metabolismo , Esclerose Múltipla/fisiopatologia , Explosão Respiratória , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
Paraoxonase, an enzyme associated with high-density lipoprotein (HDL-PON), exerts a protective effect against oxidative damage of circulating cells and lipoproteins, modulates the susceptibility of HDL to atherogenic modifications such as glycation and homocysteinylation, and even exerts an antiinflammatory role. The aim of the present study was to investigate the relationship between lipoprotein oxidative stress and the activity of HDL-PON in healthy and obese subjects. Therefore, the activity of HDL-PON and the levels of lipid hydroperoxides in HDL and low-density lipoprotein (LDL) isolated from plasma of obese females (n = 12) and age-sex-matched controls (n = 31) were compared. Our results demonstrated for the first time that the activity of HDL-PON in obese subjects was significantly lower compared with that in controls (P < 0.001). Moreover, our results showed a significant increase in the levels of lipid hydroperoxides in HDL and LDL isolated from obese subjects (P < 0.001). The negative correlations established between HDL-PON activity and the levels of lipid hydroperoxides associated with HDL and LDL confirm the relationship between paraoxonase activity and lipid peroxidation of lipoproteins. Plasma levels of leptin correlated negatively with HDL-PON activity and positively with levels of lipid hydroperoxides in HDL and LDL of obese subjects, suggesting a relationship between leptin and oxidative damage of lipoproteins. In conclusion, our study demonstrated that the increase in oxidative stress in LDL and HDL of obese subjects is associated with a decrease in HDL-PON activity. The lower paraoxonase activity and the compositional changes in HDL and LDL could contribute to the greater risk of cardiovascular disease associated with obesity.
Assuntos
Arildialquilfosfatase/metabolismo , Lipoproteínas HDL/sangue , Obesidade/metabolismo , Adulto , Peso Corporal , Feminino , Humanos , Peróxidos Lipídicos/metabolismo , Lipoproteínas LDL/sangue , Estresse Oxidativo/fisiologia , Triglicerídeos/sangueRESUMO
High-density lipoproteins (HDL) plays a key role in the protection against oxidative damage of lipoprotein and biological membranes. The aim of the present study was to investigate the relationship between the antioxidant role of HDL and the HDL-paraoxonase (PON) activity in healthy subjects and in type 1 diabetic patients. Moreover, the ability of HDL of controls and diabetic patients to protect and/or repair biological membranes from oxidative damage was studied. HDL were isolated from 31 type 1 diabetic patients and 31 sex- and age-matched healthy subjects and immediately used to evaluate lipid hydroperoxides and HDL-PON activity. Erythrocyte membranes obtained from healthy subjects were oxidized with 2,2-azo-bis(2-aminidinopropane)dihydrochloride and then incubated in the presence of HDL isolated from healthy or type 1 diabetic subjects, with measurements of membrane lipid hydroperoxides before and after the incubation. HDL from type 1 diabetic patients showed higher levels of lipid hydroperoxides and a lower activity of HDL-PON than healthy subjects. Moreover, HDL of type 1 diabetic patients protected less efficiently erythrocyte membranes against oxidative damage compared with HDL from healthy subjects. A negative correlation was found between HDL-PON activity and the levels of hydroperoxides of HDL, confirming the relationship between PON and lipid peroxidation and suggesting that subjects with low PON activity are more exposed to oxidative damage than subjects with high PON activity. The ability of HDL to protect erythrocyte membranes was positively correlated with HDL-PON activity and negatively correlated with the levels of lipid hydroperoxides of HDL of healthy subjects. These results confirm a linkage between PON activity and lipid peroxidation of lipoproteins and suggest that the ability of HDL to protect erythrocyte membranes might be related to the PON activity. It might be hypothesized that the decrease of PON activity in diabetic patients and the lower HDL protective action against membrane peroxidation could contribute to acceleration of arteriosclerosis in type 1 diabetes mellitus.
Assuntos
Arildialquilfosfatase/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Membrana Eritrocítica/enzimologia , Lipoproteínas HDL/metabolismo , Adulto , Angiopatias Diabéticas/metabolismo , Feminino , Humanos , Peroxidação de Lipídeos , Peróxidos Lipídicos/metabolismo , Masculino , Pessoa de Meia-Idade , Estresse OxidativoRESUMO
The aim of the present study was to investigate the effect exerted by low-density lipoprotein (LDL) modified by homocysteine (Hcy)-thiolactone (Hcy-LDL) on functional properties on human endothelial cells. Hcy-thiolactone, a reactive product formed in human cells from enzymatic conversion of Hcy, was hypothesized to play an important role in Hcy-induced vascular damages. Using endothelial cultured cells [human aortic endothelial cells (HAEC)] as cellular model, we evaluated nitric oxide (NO) production, cytoplasmic Ca(2+) levels, Na(+)/K(+)-ATPase activity, and peroxynitrite production in cells incubated in the presence of control LDL or Hcy-LDL. Homocysteinylation of LDL was carried out by incubation of LDL, isolated from plasma of healthy subjects, with 100 microm Hcy-thiolactone. A significant increase in cytoplasmic Ca(2+) levels and peroxynitrite production and a decrease in Na(+)/K(+)-ATPase and NO production in HAEC incubated with Hcy-LDL compared with HAEC incubated with control LDL were observed. Moreover, a positive correlation was found between Na(+)/K(+)-ATPase activity and cytoplasmic Ca(2+) content and between peroxynitrite activity and cytoplasmic Ca(2+) content. In conclusion, our results demonstrated that LDL homocysteinylated in vitro induced alterations of functional properties and NO metabolism of human endothelial cells.
Assuntos
Aorta/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Homocisteína/análogos & derivados , Homocisteína/farmacologia , Lipoproteínas LDL/farmacologia , Adulto , Aorta/metabolismo , Cálcio/metabolismo , Células Cultivadas , Células Endoteliais/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase/metabolismo , Ácido Peroxinitroso/biossíntese , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
The interaction between low density lipoproteins (LDL) and platelets might play a central role in the development of atherosclerosis in diabetes. The aim of the present study was to investigate whether the glycation of LDL is associated with modifications of their physico-chemical and functional properties and to study the action of glycated LDL (glycLDL) on platelets. LDL and platelets were isolated from 15 healthy subjects. The content of thiobarbituric acid-reactive substances and the generalized polarization of the fluorescent probe Laurdan were determined in LDL glycated in vitro. Platelets were incubated with native LDL, GlycLDL, and minimally oxidized LDL, and the following parameters were evaluated: platelet aggregation, nitric oxide production, intracellular Ca(2+) concentrations, Na(+)/K(+)-adenosine triphosphatase (Na(+)/K(+)-ATPase), and Ca(2+)-ATPase activities. GlycLDL showed increased thiobarbituric acid-reactive substance levels, a red shift of the Laurdan emission maximum, and a decrease in generalized polarization, indicating a higher polarity and a reduced molecular order compared with native LDL. GlycLDL caused a significant increase in platelet nitric oxide production, intracellular Ca(2+) concentration, and aggregating response to ADP; an inhibition of the platelet membrane Na(+)/K(+)-ATPase activity; and a stimulation of Ca(2+)-ATPase activity. Minimally oxidized LDL did not cause statistically significant changes in the parameters studied. The present work demonstrates that glycation induces compositional and structural changes in LDL and suggests that an altered interaction between glycLDL and platelets might play a role in the vascular complications of diabetes.
Assuntos
2-Naftilamina/análogos & derivados , Plaquetas/efeitos dos fármacos , Plaquetas/fisiologia , Lipoproteínas LDL/farmacologia , Adulto , Polaridade Celular/efeitos dos fármacos , Corantes Fluorescentes , Glucose/farmacologia , Produtos Finais de Glicação Avançada , Humanos , Lauratos , Lipoproteínas LDL/química , Masculino , Agregação Plaquetária/efeitos dos fármacos , Substâncias Reativas com Ácido Tiobarbitúrico/análiseRESUMO
Several studies have demonstrated that the isoflavone genistein exerts a protective effect against lipid peroxidation of low density lipoproteins (LDL). Aim of our study was to investigate whether genistein protects high density lipoproteins (HDL), isolated from normolipemic subjects, against Cu(++)-induced lipid peroxidation. Our results demonstrated that genistein exerts an inhibitory effect against Cu(++)-induced lipid peroxidation of HDL, as shown by the lower increase in the levels of conjugated dienes in lipoproteins oxidized after preincubation with different concentrations of genistein (0.5-2.5microM). Moreover the analysis of fluorescence emission spectra of tryptophan (Trp) and Laurdan (6-dodecanoyl-2-dimethyl-aminonaphthalene) demonstrated that genistein prevents the alterations of apoprotein structure and physico-chemical properties, associated with Cu(++)-triggered lipid peroxidation of lipoproteins. The protective effect exerted by genistein against oxidative damage of lipoproteins was realized at concentrations similar to those observed in plasma of human subjects consuming a traditional soy diet or receiving a soy supplement. Therefore, we suggested that antioxidant activity exerted by genistein against lipid peroxidation of HDL in vitro could be of physiological relevance.
Assuntos
2-Naftilamina/análogos & derivados , Antioxidantes/farmacologia , Cobre/farmacologia , Genisteína/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Lipoproteínas HDL/metabolismo , 2-Naftilamina/análise , Apoproteínas/química , Cobre/antagonistas & inibidores , Fluorescência , Humanos , Técnicas In Vitro , Lauratos/análise , Alimentos de Soja , Triptofano/análiseRESUMO
We investigated the effect of homocysteine (Hcy)-thiolactone on the activity of the enzyme paraoxonase (PON) associated with human high-density lipoprotein (HDL-PON). HDL were isolated from plasma of normolipidemic subjects. The increase in the levels of sulfhydryl groups (-SH) in HDL incubated with Hcy-thiolactone demonstrates that homocysteinylation of HDL occurs. The increase of -SH groups correlated with the basal values of HDL-PON activity (r = -0.73, P <.001, and r = -0.70, P <.002 using 10 micromol/L and 1 mmol/L Hcy-thiolactone, respectively) suggesting a relationship between the susceptibility of HDL to homocysteinylation and HDL-PON activity. A decrease in the activity of the enzyme HDL-PON was observed in homocysteinylated HDL (Hcy-HDL). The negative correlation established between the basal levels of HDL-PON activity and the percentage decrease of HDL-PON activity (r = -0.76, P <.001, and r = -0.86, P <.001 using 10 micromol/L or 1 mmol/L Hcy-thiolactone, respectively) suggests that subjects with higher HDL-PON activity have a lower decrease in PON activity with respect to subjects with lower HDL-PON activity. The positive correlation established between the percentage decrease of PON activity and the percentage increase of -SH groups in Hcy-HDL (r = 0.80, P <.001, and r = 0.76, P <.001 in HDL incubated in the presence of 10 micromol/L and 1 mmol/L Hcy-thiolactone, respectively) suggests that the modifications of HDL-PON activity are likely related to the compositional changes at the lipoprotein surface of Hcy-HDL. The enzyme PON contributes to the protective role of HDL against the oxidative damage and against toxicity exerted by Hcy involved in the development of atherosclerosis. Therefore the significant decrease of the enzyme activity in HDL incubated with Hcy-thiolactone suggests that homocysteinylation could render HDL less protective against oxidative damage and against toxicity of Hcy-thiolactone.
Assuntos
Esterases/sangue , Homocisteína/análogos & derivados , Homocisteína/farmacologia , Lipoproteínas HDL/sangue , Adulto , Arildialquilfosfatase , Homocisteína/metabolismo , Humanos , Lipoproteínas HDL/química , Lipoproteínas HDL/metabolismo , Masculino , Compostos de Sulfidrila/análiseRESUMO
We investigated the effect of aluminium (Al3+) on lipid peroxidation and physico-chemical properties of high density lipoproteins (HDL) isolated from human plasma. Our results demonstrated that Al3+ enhances lipid peroxidation of human HDL as shown by the significant increase in lipid hydroperoxides in Al-treated HDL with respect to control HDL. The oxidative effect was higher at acid pH (pH 5.5) with respect to pH 7.4. Moreover, a stimulating effect of Al3+ on iron-induced lipid peroxidation of HDL was demonstrated. The study of the effect of Al3+ on the physico-chemical properties of HDL, using the fluorescence polarization (Pf) of the probes TMA-DPH (1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene iodide) and DPH (1,6-diphenyl-1,3,5-hexatriene), showed a significant decrease of Pf in Al-treated HDL with respect to control. These results suggest that Al3+ induces a decrease of molecular order at the lipoprotein surface. Moreover, the study of tryptophan (Trp) fluorescence demonstrated that aluminium induces structural modifications of HDL apoproteins and on HDL physico-chemical properties. The effect of Al3+ on lipid peroxidation of HDL was observed at aluminium concentrations similar to those observed in the brain of patients affected by neurological diseases. Aluminium-induced oxidative damage of HDL could be involved in the development of neurological diseases.
Assuntos
Alumínio/farmacologia , Difenilexatrieno/análogos & derivados , Peroxidação de Lipídeos/efeitos dos fármacos , Lipoproteínas HDL/metabolismo , Difenilexatrieno/química , Relação Dose-Resposta a Droga , Compostos Ferrosos/farmacologia , Polarização de Fluorescência , Corantes Fluorescentes/química , Humanos , Concentração de Íons de Hidrogênio , Peróxidos Lipídicos/metabolismo , Lipoproteínas HDL/sangue , Oxirredução/efeitos dos fármacos , Espectrometria de Fluorescência , Substâncias Reativas com Ácido Tiobarbitúrico/química , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Triptofano/químicaRESUMO
Recent studies have demonstrated that Apo AIV exerts a protective effect against atherosclerosis. Moreover, Qin et al. (Am. J. Physiol. 274 (1998) H1836) have demonstrated that Apo AIV, isolated from rat plasma, exerts an inhibitory effect against Cu(2+)-induced lipid peroxidation of intestinal lymph and LDL. The aim of the study was to investigate whether human Apo AIV exerts a protective effect against Cu(2+)-induced lipid peroxidation. Our results demonstrated that human Apo AIV exerted an inhibitory effect against Cu(2+) and AAPH induced lipid peroxidation of VLDL, as shown by the lower increase in the levels of TBARS and conjugated dienes in lipoproteins preincubated with Apo AIV. In addition, the tryptophan (Trp) and probe 2-(dimethylamino)-6-lauroylnaphthalene (Laurdan) fluorescence studies demonstrated that the modifications of spectral properties in both lipoproteins preincubated with Apo AIV were lower with respect to ox-lipoproteins, suggesting that Apo AIV prevents the modification of physico-chemical properties due to peroxidation.
Assuntos
2-Naftilamina/análogos & derivados , Apolipoproteínas A/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Lipoproteínas VLDL/metabolismo , Amidinas/farmacologia , Cobre/farmacologia , Fluorescência , Corantes Fluorescentes , Humanos , Lauratos , Oxidantes/farmacologia , Triptofano/químicaRESUMO
We investigated the effect of incubation of high density lipoprotein (HDL) under hyperglycaemic conditions on lipid composition, physicochemical properties and activity of paraoxonase (PON), a calcium-dependent enzyme associated with HDL that contributes to the antiatherogenicity of this lipoprotein. HDL incubated for three days with various glucose concentrations (0-100 mM) had significant increases in thiobarbituric acid-reactive substances (TBARS) and conjugated dienes with respect to control HDL. These results suggest that lipid peroxidation accompanies HDL glycation in vitro. The susceptibility to lipid peroxidation was higher in HDL isolated from subjects with low HDL-paraxonase activity with respect to subjects with higher HDL-PON activity. The lipid compositional changes were associated with modifications of apoprotein conformation as shown by the red-shifted position of the maximum emission of tryptophan in treated HDL. The decrease in the Gp (generalized polarization) value and the red-shifted position of the maximum emission of Laurdan incorporated in treated HDL demonstrate modifications of order and polarity with respect to control HDL. The negative correlation established between the Gp value and TBARS demonstrates that the modifications in molecular order are likely related to the increase in lipid peroxidation products. The activity of paraoxonase was significantly decreased in HDL incubated at 37 degrees C; a greater decrease occurred in the presence of 50 mM and 100 mM glucose. This study demonstrates modifications of lipid composition, apoprotein conformation and physicochemical properties of HDL incubated in the presence of glucose. These modifications affect the activity of HDL-associated paraoxonase. The physicochemical properties of lipoproteins play a regulatory role in lipoprotein function. The modification of order and polarity of glycated HDL and the alterations in paraoxonase activity could potentially contribute to the accelerated atherosclerosis in diabetic patients.
Assuntos
2-Naftilamina/análogos & derivados , Esterases/metabolismo , Glucose/metabolismo , Lipoproteínas HDL/metabolismo , 2-Naftilamina/química , Arildialquilfosfatase , Corantes Fluorescentes/química , Humanos , Lauratos/química , Peroxidação de Lipídeos/fisiologia , Lipoproteínas HDL/química , Microscopia de Fluorescência , Substâncias Reativas com Ácido Tiobarbitúrico/química , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
The aim of this study was to inquire the antioxidant status in plasma and lipoproteins isolated from normal subjects possessing different ApoE genotypes. For this purpose we investigated blood samples from 106 healthy blood donors: the distribution of ApoE alleles (E2/E2 = 0.9%, E2/E3 = 10.4%, E2/E4 = 2.8%, E3/E3 = 71.7%, E3/E4 = 12.3% and E4/E4 1.9% with 1, 11, 3, 76, 13, and 2 subjects respectively for each genotype) was in agreement with previous data. Almost no differences were found in the concentrations of both coenzyme Q10 (CoQ10) and vitamin E for the different genotypes. Concentration of CoQ10 in isolated lipoproteins was also similar, in the different genotypes, when referred to cholesterol; CoQ10 in LDL was higher for the E3/E3 subjects when referred to protein. Neither CoQ10 nor vitamin E correlated with paraoxonase (PON) activity or cholesteryl-ester hydroperoxides (CHP). Furthermore, there was no correlation between the same lipophilic antioxidants and CHP levels. The only E2 homozygous subject found had high levels of PON and low levels of CHP; the two E4/E4 subjects had low PON activity together with low levels of CHP.
Assuntos
Antioxidantes/análise , Apolipoproteínas E/genética , Ubiquinona/análogos & derivados , Ubiquinona/sangue , Alelos , Apolipoproteína E2 , Apolipoproteína E3 , Apolipoproteína E4 , Apolipoproteínas E/sangue , Arildialquilfosfatase/sangue , Doadores de Sangue , Ésteres do Colesterol/sangue , Coenzimas , Genótipo , Humanos , Peróxido de Hidrogênio/sangue , Lipoproteínas HDL/sangue , Lipoproteínas LDL/sangue , Vitamina E/sangueRESUMO
The objective of the present paper is to evaluate the post-prandial response to some varieties of gluten free (GF) pasta that are commonly consumed in Italy. The glycaemic responses were compared with a glucose standard in healthy subjects and gluten-free diet celiac subjects. Subjects were served portions of the test foods and a standard food (glucose), on separate occasions, each containing 50 g available carbohydrates. Capillary blood glucose was measured from finger-prick samples in fasted subjects and at 15, 30, 45, 60, 90 and 120 minutes after the consumption of each test food. For each type of pasta, the glycaemic index (GI) was calculated by expressing the incremental area under the blood glucose curve as a percentage of each subject's average incremental area under the blood glucose curve (AUC) for the standard food. Gluten free pasta exhibited a range of GI values from 46 to 66. The glycaemic load (GL) and glycaemic profile (GP) were also calculated. A higher GI value was observed in pasta containing rice flour as the main ingredient. Lower values were observed in pasta obtained using corn or a mixture of corn and rice flour as the main ingredients. The results were confirmed in celiac subjects. The information presented in this paper may be useful in helping celiac people to select low-GI pasta.
Assuntos
Glucose/administração & dosagem , Glutens/administração & dosagem , Período Pós-Prandial , Adulto , Glicemia/metabolismo , Estudos de Casos e Controles , Doença Celíaca/dietoterapia , Dieta Livre de Glúten , Feminino , Farinha/análise , Índice Glicêmico , Voluntários Saudáveis , Humanos , Itália , Masculino , Refeições , Oryza/químicaRESUMO
OBJECTIVE: Obesity and/or psychopathological disorders of parents represent risk factors for childhood obesity. The aim of the study was to investigate the link between obesity in pregnancy and oxidative stress. METHODS: Venous blood was collected from 37 women at the eighth month of gestation (19 obese and 28 normal weight). Cord blood was obtained at birth from newborns of obese mothers and controls. Cord blood and maternal blood was used to separate plasma to be used for the evaluation of leptin, oxidized LDL and paraoxonase (PON1) activity. RESULTS: Higher levels of leptin were observed both in maternal blood and cord blood of children of obese women compared to normal-weight women. The data also showed lower levels of PON1 activity in plasma of obese women and in the cord blood of their children. Furthermore, a positive correlation was established between levels of PON1 activity in maternal blood and cord blood, suggesting a relationship between PON1 in maternal plasma and fetal cord blood. CONCLUSIONS: Essential obesity in pregnancy is associated with hyperleptinemia. PON1 exerts an antioxidant role; therefore, our results demonstrated that obesity exposes to an increased susceptibility to oxidative damage in both mothers and newborns.
Assuntos
Arildialquilfosfatase/sangue , Leptina/sangue , Obesidade/sangue , Complicações na Gravidez/sangue , Adulto , Estudos de Casos e Controles , Feminino , Sangue Fetal/enzimologia , Humanos , Estresse Oxidativo , GravidezRESUMO
BACKGROUND: Homocysteine (Hcy) is an independent risk factor for cardiovascular disease (CVD). Individuals with Type 1 and Type 2 diabetes are more susceptible to the effects of homocysteine than non-diabetic subjects. The interaction between homocysteine-thiolactone (Hcy-thiolactone), a reactive product of Hcy, and low-density lipoproteins (LDL) induces the formation of homocystamide-LDL adducts (Hcy-LDL) and it has been suggested that homocysteinylation could increase atherogenicity of lipoproteins. AIM: The aim of the study was to compare the effect of in vitro homocysteinylation of LDL isolated from healthy control subjects (C-LDL) and from Type 1 diabetic patients (DM-LDL) and to investigate the effect of homocysteinylated LDL (Hcy-C-LDL and Hcy-DM-LDL) on peroxynitrite production of endothelial cells. METHODS: The in vitro homocysteinylation of LDL isolated from control (n = 12) and DM subjects (n = 12) was carried out by incubating lipoproteins with Hcy-thiolactone. The reaction was verified by quantifying the increase in sulphydryl groups (-SH groups) in Hcy-LDL with respect to control LDL. Control and homocysteinylated LDL were incubated with human aortic endothelial cells (HAEC) in culture. Peroxynitrite production in cells treated in different experimental conditions was assayed by a fluorimetric method. RESULTS: The increase in -SH groups after incubation with homocysteine was greater in LDL from diabetic subjects compared with LDL from control subjects (P < 0.001). In addition, peroxynitrite production from HAEC incubated with Hcy-LDL from diabetic patients was greater than after incubation with Hcy-LDL from control subjects and untreated LDL from diabetic patients (P < 0.001). CONCLUSIONS: These results show that LDL from diabetic patients is more susceptible to in vitro homocysteinylation than LDL from non-diabetic individuals and demonstrate that the compositional changes in Hcy-LDL from diabetic subjects have cytotoxic effects on human endothelial cells.
Assuntos
Aterosclerose/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Células Endoteliais/metabolismo , Lipoproteínas LDL/metabolismo , Ácido Peroxinitroso/biossíntese , Adulto , Aorta/metabolismo , Aterosclerose/complicações , Diabetes Mellitus Tipo 1/complicações , Homocisteína/análogos & derivados , Homocisteína/farmacologia , Humanos , Lipoproteínas LDL/efeitos dos fármacos , Masculino , Protetores contra Radiação/farmacologiaRESUMO
Paraoxonase, an enzyme associated with high density lipoproteins (HDL), plays an important role in the anti-oxidant and anti-inflammatory properties exerted by HDL. Increasing evidence supports a role of free radicals and oxidative stress in the inflammatory processes and in the pathogenesis of multiple sclerosis (MS). The aim of this study was to further investigate the relationship between oxidative damage and MS; therefore we compared the paraoxonase activity and levels of cholesteryl ester hydroperoxides (CE-OOH), as marker of lipid peroxidation, in plasma isolated from healthy subjects (n = 89) and from MS patients (n = 24) in the early stage disability (EDSS<3.5). Our results demonstrated for the first time that the activity of paraoxonase in the plasma of MS subjects was significantly lower with respect to controls (P <0.001). Moreover, our results showed a significant increase in the levels of CE-OOH in plasma from MS subjects (P<0.001). CE-OOH are biologically active substances derived from the oxidation of cholesteryl ester localized in the hydrophobic core of plasma lipoproteins (HDL, LDL). Therefore, our study demonstrates alterations of lipoprotein peroxidation in MS and provides further evidence that oxidative stress and impairment of the anti-oxidant system may play a role in MS.
Assuntos
Arildialquilfosfatase/metabolismo , Peroxidação de Lipídeos , Peróxidos Lipídicos/sangue , Esclerose Múltipla/metabolismo , Adulto , Biomarcadores/sangue , Ésteres do Colesterol/sangue , Feminino , Humanos , Lipídeos/sangue , Masculino , Esclerose Múltipla/imunologia , Neurite (Inflamação)/imunologia , Neurite (Inflamação)/metabolismo , Estresse OxidativoRESUMO
BACKGROUND AND AIM: Several studies have shown that non-enzymatic glycation and oxidative damage play an important role in the pathogenesis of neurological diseases. Increased levels of advanced glycation end-products (AGEs) and of lipid peroxidation products have been observed in the brain, in the cerebrospinal fluid (CSF) and in the plasma of subjects affected by Alzheimer's disease (AD). The aim of this study was to investigate the effect of non-enzymatic glycation on aluminium-induced lipid peroxidation and on the stimulatory effect exerted by aluminium on iron-triggered oxidation of high density lipoproteins (HDL) isolated from human plasma. METHODS AND RESULTS: Aluminium (10-200 microM) and iron (20 microM) induced a significant increase in lipid hydroperoxides in HDL compared to untreated HDL. Therefore, our results confirm that aluminum and iron exert an oxidant effect on HDL. Moreover, aluminium exerted a stimulatory effect on iron-induced lipid peroxidation of HDL, in agreement with our previous studies. The aluminum/iron-induced increase in lipid hydroperoxides was significantly higher in HDL incubated for different time periods (24-72 hours) in the presence of 50 mM glucose (Gly-HDL) compared to HDL incubated alone. These results demonstrate that Gly-HDL is more susceptible to aluminium and iron-oxidative treatment with respect to control HDL. CONCLUSION: We suggest that aluminium and iron-induced oxidative damage on HDL could be involved in the development of neurological diseases and that glycation of HDL could represent an additional risk factor for these human diseases.