RESUMO
The increment of eryptosis in lead-exposed workers has been associated with oxidative stress, having as the main mediator [Ca2+]i. However, other molecules could participate as signals, such as PLA2 and SMase, which have been proposed to increase PGE2 and ceramides, both involved in the increment of PS externalization due to osmotic stress. To study the role of these enzymes in lead intoxication, we studied 30 lead exposed workers and 27 non-lead exposed individuals. We found, compared to non-exposed subjects, lead intoxication characterized by high blood lead concentration (medianâ¯=â¯39.1⯵g/dL), and low δ-ALAD activity (medianâ¯=â¯348â¯nmol of porphobilinogen/h/mL); oxidative stress with high lipid peroxidation (medianâ¯=â¯1.31â¯nmol of malondialdehyde/mL) and low TAC (medianâ¯=â¯370â¯mM Trolox equivalents); a higher enzymatic activity of PLA2 (medianâ¯=â¯518 AFU/mg) and SMase (medianâ¯=â¯706 AFU/mg) and higher eryptosis (medianâ¯=â¯0.92% PS externalization). Correlation and conditional probability analyses permit to associate oxidative stress and eryptosis with high PLA2 activity. However, high SMase activity was only associated with PLA2 activity. The role of these enzymes in the signal path to eryptosis induced by oxidative stress in lead-exposed workers is discussed.
Assuntos
Poluentes Ambientais/efeitos adversos , Eriptose/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Intoxicação por Chumbo/etiologia , Chumbo/efeitos adversos , Exposição Ocupacional/efeitos adversos , Estresse Oxidativo/efeitos dos fármacos , Fosfolipases A2/sangue , Esfingomielina Fosfodiesterase/sangue , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Poluentes Ambientais/sangue , Eritrócitos/enzimologia , Eritrócitos/patologia , Humanos , Chumbo/sangue , Intoxicação por Chumbo/sangue , Intoxicação por Chumbo/enzimologia , Intoxicação por Chumbo/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Pessoa de Meia-Idade , Sintase do Porfobilinogênio/sangue , Medição de Risco , Transdução de Sinais , Adulto JovemRESUMO
OBJECTIVES: Expression of phospholipids and related molecules could provide panels of multiple biomarkers searching for the signature of Alzheimer's disease (AD). The aim of the present study was to quantify ten phospholipids and simultaneously determine phospholipase A2 (PLA2) activity in blood of mild cognitive impairment (MCI) and AD patients. METHODS: Thirty-four AD, 20 MCI and 25 controls were enrolled. The phospholipids where analysed using the AbsoluteIDQ® p180 Kit. PLA2 activities were accessed in platelets by a radio-enzymatic assay. RESULTS: The study failed to fix the ten phospholipids as a panel to predict AD; the levels of PCaaC36:6, PCaaC40:6 and C16:1-OH were lower in MCI than in controls (P = 0.041, P = 0.012, P = 0.044 respectively). PCaaC40:2 levels were lower in MCI than in AD (P = 0.041). The converters MCI-AD showed at baseline lower levels of PCaaC40:2 (P = 0.050) and PCaaC40:6 (P = 0.037) than controls. iPLA2 activity was reduced in AD and MCI than in controls (P < 0.001). We found positive correlation in the control group between PCaaC38:6 and tPLA2 (r = 0.680; P = 0.001) and sPLA2 (r = 0.601; P = 0.004); PCaaC40:1 and iPLA2 (r = 0.503; P = 0.020); PCaaC40:6 and tPLA2 (r = 0.532; P = 0.013) and sPLA2 (r = 0.523; P = 0.015). CONCLUSIONS: Lipids metabolites in plasma might indirectly indicate changes in neuronal membrane and this deregulation can outline the transition between healthy and diseased brains.
Assuntos
Doença de Alzheimer/enzimologia , Disfunção Cognitiva/enzimologia , Fosfolipases A2/sangue , Fosfolipídeos/sangue , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/sangue , Biomarcadores/sangue , Plaquetas/enzimologia , Estudos de Casos e Controles , Disfunção Cognitiva/sangue , Feminino , Humanos , Metabolismo dos Lipídeos , Masculino , Testes NeuropsicológicosRESUMO
Fluidity is an important neuronal membrane property and it is influenced by the concentration of polyunsaturated fatty acids (PUFAs) in membrane phospholipids. Phospholipase A(2) (PLA(2)) is a key enzyme in membrane phospholipid metabolism, generating free PUFAs. In Alzheimer disease (AD), reduced PLA(2) activity, specifically of calcium-dependent cytosolic PLA(2) (cPLA(2)) and calcium-independent intracellular PLA(2) (iPLA(2)), and phospholipid metabolism was reported in the frontal cortex and hippocampus. This study investigated the effects of in vivo infusion of the dual cPLA(2) and iPLA(2) inhibitor MAFP into rat brain on PLA(2) activity and membrane fluidity parameters in the postmortem frontal cortex and dorsal hippocampus. PLA(2) activity was measured by radioenzymatic assay and membrane fluidity was determined by fluorescence anisotropy technique using three different probes: DPH, TMA-DPH, and pyrene. MAFP significantly inhibited PLA(2) activity, reduced the flexibility of fatty acyl chains (indicated by increased DPH anisotropy), increased the fluidity in the lipid-water interface (indicated by decreased TMA-DPH anisotropy), and increased the lipid lateral diffusion in the hydrocarbon core (represented by pyrene excimer formation) of membranes in both brain areas. The findings suggest that reduced cPLA(2) and iPLA(2) activities in AD brain might contribute to the cognitive impairment, in part, through alterations in membrane fluidity parameters.
Assuntos
Encéfalo/fisiologia , Fluidez de Membrana/fisiologia , Fosfolipases A2/metabolismo , Doença de Alzheimer/metabolismo , Animais , Ácidos Araquidônicos/farmacologia , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Avaliação Pré-Clínica de Medicamentos , Inibidores Enzimáticos/farmacologia , Humanos , Masculino , Fluidez de Membrana/efeitos dos fármacos , Organofosfonatos/farmacologia , Inibidores de Fosfolipase A2 , Fosfolipases A2/sangue , Ratos , Ratos WistarRESUMO
Plasma phospholipases A(2) (PLA(2)) hydrolyze phospholipids of circulating lipoproteins or deposited in arteries producing bioactive lipids believed to contribute to the atherosclerotic inflammatory response. PLA(2)(s) are elevated in obesity and type 2 diabetes (T2D) but it is not clear which of these conditions is the cause since they frequently coexist. This study attempts to evaluate if high plasma PLA(2)(s) activities and markers of their effects in lipoproteins are associated with obesity or T2D diabetes, or with both. Total PLA(2) and Ca(2+)-dependent and -independent activities, lipids, lipoproteins, apoAI, and apoB apolipoproteins and affinity of apoB-lipoproteins for arterial proteoglycans were measured, as well as Inflammation markers. These parameters were evaluated in plasma samples of four groups: (i) apparently healthy controls with normal BMI (nBMI), (ii) obese subjects with no T2D, (iii) patients with T2D but with nBMI, and (iv) obese patients with T2D. PLA(2) activities were measured in the presence and absence of Ca(2+) and in the presence of specific inhibitors. Obese subjects, with or without T2D, had high activities of total PLA(2) and of Ca(2+)-dependent and Ca(2+)-independent enzymes. The activities were correlated with inflammation markers in obese subjects with and without diabetes and with alterations of low-density lipoproteins (LDLs) that increased their affinity for arterial proteoglycans. Ca(2+)-dependent secretory (sPLA(2)) enzymes were the main responsible of the obesity-associated high activity. We speculate that augmented PLA(2)(s) activity that increases affinity of circulating LDL for arterial intima proteoglycans could be another atherogenic component of obesity.
Assuntos
LDL-Colesterol/sangue , Diabetes Mellitus Tipo 2/sangue , Mediadores da Inflamação/sangue , Obesidade/sangue , Fosfolipases A2/sangue , Proteoglicanas/sangue , Adulto , Idoso , Biomarcadores/sangue , Cálcio/metabolismo , Diabetes Mellitus Tipo 2/complicações , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Obesidade/complicações , Valores de ReferênciaRESUMO
BACKGROUND AND AIMS: Many studies suggest that the different steps of the atherosclerotic process may be mediated by metalloproteases (MMPs). MMP-9 and MMP-2, which are highly expressed in the vulnerable regions of the atherosclerotic plaques, have been suggested to be causally involved in plaque rupture. In another manner linked with LDL, lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) hydrolyzes phospholipids generating proinflammatory and proatherogenic products. Our aim was to evaluate plasma activity of MMP-2 and 9, as well as Lp-PLA(2), in subjects with coronary artery stenosis in comparison with controls and to correlate these activities with lipoprotein profile and general biomarkers of inflammation. METHODS: Forty two subjects who had undergone coronary angiography were divided into two groups: patients with coronary vessels with at least 45% stenosis (CAD [coronary artery disease], n = 24) and patients without angiographically detectable coronary artery disease (controls, n = 18). Plasma activity of MMP-2 and MMP-9 was measured and correlated with markers of systemic inflammation (hs-CRP), subendothelial inflammation (Lp-PLA(2)) and lipoprotein profile. RESULTS: Plasma activity of both MMPs was consistently higher in patients than in controls (p <0.01). Pro-MMP-2 (r = 0.34, p <0.01) and MMP-9 (r = 0.51, p <0.02) activities correlated with apoprotein B. Pro-MMP-2 correlated with hs-CRP (r = 0.47, p <0.01) and inversely with HDL cholesterol (r = -0.35, p <0.02). No differences were observed in Lp-PLA(2) between patients and controls (15.2 +/- 4.0 vs. 15.4 +/- 4.5 micromol/mL/h, p = NS, respectively), and no correlation was observed with MMPs. CONCLUSIONS: MMP activity was higher in CAD than in controls. The correlation observed between pro-MMP-2 and high-sensitive C-reactive protein (hs-CRP) may be due to specific systemic inflammatory processes. No correlation was observed between Lp-PLA(2) and MMPs.