In this study, we focused on an analysis of biguanides effects on mitochondrial enzyme activities, mitochondrial membrane potential and membrane permeabili ty transition pore function. We used phenformin, which is more efficient than metformin, and evaluated its effect on rat liver mitochondria and isolated hepatocytes. In contrast to prev iously published data, we found that phenformin, after a 5 min pr e-incubation, dose-dependently inhibits not only mitochondrial complex I but also complex II and IV activity in isolated mitochondria. The enzymes complexes inhibition is paralleled by the decreased respiratory control index and mitochondrial membrane potent ial. Direct measurements of mitochondrial swelling revealed that phenformin increases the resistance of the permeability transition pore to Ca 2+ ions. Our data might be in agreement with the hypothesis of Schäfer (1976) that binding of biguanides to membrane phospholipids alters membrane properties in a non-specific manner and, subsequently, different enzyme activities are modified via lipid phase. However, our measurements of anisotropy of fluorescence of hydrophobic membrane probe diphenylhexatriene have not shown a measurable effect of membrane fluidity with the 1 mM concentration of phenformin that strongly inhibited complex I activity. Our data therefore suggest that biguanides could be considered as agents with high efficacy but low specifity., Z. Drahota ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
The effect of low (1 mg/animal/day), medium (10 mg/animal/day) and high (100 mg/animal/day) intake of ascorbic acid on tissue lipid peroxidation (LPO) and the physical state of erythrocyte membranes was investigated in female guinea-pigs fed a vitamin E low diet. Animals were killed after 9-11 weeks and the blood, liver, lungs, kidneys and adrenals were analysed. The LPO was estimated by the determination of malondialdehyde with HPLC. The physical state of erythrocyte membranes was determined spectrofluorometrically and expressed as fluorescence polarization of membrane lipid specific probe 1,6-diphenyl-1,3,5-hexatriene. The LPO concentrations in the liver and adrenals of the group on a low vitamin C intake were significantly increased. A significant non-linear negative correlation between C vitamin levels and LPO concentrations was found in these tissues. The fluidity of erythrocyte membranes as a measure of their structural state was significantly lower in the group with a low intake of C vitamin. It is probable that the water-soluble antioxidants, such as vitamin C, act in the plasma as primary defense against oxidative stress if the radicals are formed initially in the aqueous phase of whole blood.
Membrane fluidity is a widely recognized biophysical variable that provides information about structural organization of the subcellular membranes exhibiting physical characteristics of liquid crystals. The term “fluidity” reflects in this case the tightness in packing of acyl parts of the membrane phospholipid molecules, a feature that may influence considerably the molecular mobility and via that also the sensitivity and reactivity of membranebound transporters, receptors and enzyme systems. Data presented in this review are aimed to demonstrate the substantial role of changes in membrane fluidity occurring in the processes associated with endogenous protection observed in cardiac sarcolemma and mitochondria in diverse pathologies, particularly in diabetes and hypertension., A. Ziegelhöffer, ... [et al.]., and Obsahuje seznam literatury
Acute streptozotocin diabetes mellitus (DM) as well as remote ischemic preconditioning (RPC) has shown a favorable effect on the postischemic-reperfusion function of the myocardium. Cardioprotective mechanisms offered by these experimental models involve the mitochondria with the changes in functional properties of membrane as the end-effector. The aim was to find out whether separate effects of RPC and DM would stimulate the mechanisms of cardioprotection to a maximal level or whether RPC and DM conditions would cooperate in stimulation of cardioprotection. Experiments were performed on male Wistar rats divided into groups: control, DM, RPC and DM treated by RPC (RPC+DM). RPC protocol of 3 cycles of 5-min hind limb ischemia followed by 5-min reperfusion was used. Ischemicreperfusion injury was induced by 30-min ischemia followed by 40-min reperfusion of the hearts in Langendorff mode. Mitochondria were isolated by differential centrifugation, infarct size assessed by staining with 1 % 2,3,5-triphenyltetrazolium chloride, mitochondrial membrane fluidity with a fluorescent probe DPH, CoQ9 and CoQ10 with HPLC. Results revealed that RPC as well as DM decreased the infarct size and preserved mitochondrial function by increasing the mitochondrial membrane fluidity. Both used models separately offered a sufficient protection against ischemic-reperfusion injury without an additive effect of their combination., M. Ferko, I. Kancirová, M. Jašová, I. Waczulíková, S. Čarnická, J. Kucharská, O. Uličná, O. Vančová, M. Muráriková, T. Ravingerová, A. Ziegelhöffer., and Obsahuje bibliografii
Our previous preliminary results pointed to possible seasonal variations in Mg2+-ATPase activity of rat heart mitochondria (MIT). It is not too surprising since seasonal differences were already reported in myocardial function, metabolism and ultrastructure of the intact as well as hemodynamically overloaded rabbit hearts and also in other tissues. The present study is aimed to elucidate whether seasonal differences observed in rat heart MIT Mg2+-ATPase activity will be accompanied with changes in membrane fluidity and in the content of conjugated dienes (CD) in the lipid bilayers of MIT membranes as well as whether the above seasonal differences will also be present in the diabetic heart. Our results revealed that values of Mg2+-ATPase activity in the winter/spring-period (W/S-P) exceeded significantly (p<0.05-0.001) those in the summer/autumn-period (S/A-P). Similar trend was also observed in hearts of animals with acute (8 days) streptozotocin diabetes. With the exception of values of CD in the S/A-P, all values of Mg2+-ATPase activities, membrane fluidity and CD concentrations in diabetic hearts exceeded those observed in the healthy hearts. Our results indicate that seasonal differences may play a decisive role in the evaluation of properties and function of rat heart MIT., J. Mujkošová, M. Ferko, P. Humeník, I. Waczulíková, A. Ziegelhöffer., and Obsahuje bibliografii a bibliografické odkazy
Lipid peroxidation of rat cerebral cortex membranes was induced by Fe2+/ADP and ascorbate. The rate of Na+/K+-ATPase inhibition was correlated with the increase of thiobarbituric acid-reactive substances (TBARS) and conjugated dienes (CD) and with membrane fluidity changes. Our data showed that membrane fluidity changes (evaluated by fluorescence steady-state anisotropy measurements) can participate in Na+/K+-ATPase inhibition during the initial period of lipid peroxidation process, whereas during the following period the enzyme inhibition correlates only with TBARS and CD production., H. Rauchová, Z. Drahota, J. Koudelová., and Obsahuje bibliografii
Seedlings of Cyamopsis tetragonoloba were grown on Petri dishes either in water or water plus 3 % PEG-6000 to induce water stress. The senescing cotyledons experiencing the stress exhibited loss in contents of leaf proteins and chlorophyll (Chl) and a decline in oxygen evolution. The effect of PEG treatment was more pronounced at moderate (MI) than low (LI) irradiance. The stress-induced loss in the activity of superoxide dismutase and increase in the thylakoid lipid peroxidation accompanied a change in the physical status of the bilayer membrane as demonstrated by an enhancement of room temperature Chl a fluorescence polarization and decrease in energy transfer efficiency in pigment assembly. This resulted in a sustained decrease in photosystem 2 activity blocking channels of energy utilization. The absorbed quanta, thus unutilized, were excess even at MI, leading to photoinhibitory response. and P. M. Deo, U. C. Biswal, B. Biswal.