Wire myograph is a device for the in vitro investigation of both, active and passive properties of arteries. Arteries from a variety of animal species, pathological states, and vascular beds were investigated using this method. We focus on the normalization procedure which is aimed to standardize experimental settings and, in part, to simulate physiological conditions. During normalization, it is determined the internal circumference of a vessel stretched to a tension that corresponds to the transmural pressure of 100 mm Hg (IC100). Once it is determined, the internal circumference is traditionally set to (0.9 ⋅ IC100). However, this constant 0.9, called also the normalization factor (NF), was experimentally determined for rat small mesenteric arteries only. Therefore, the aim of our work was to show the influence of different NFs on the passive tension and reactivity of both, rat femoral arteries (FA) an d the first branches of superior mesenteric arteries (MA). We found out that the maximal active wall tension of the FA was achieved at the NF value of 1.1, and that of the MA at 0.9. Considering the values of the active wall tension we suggest that higher reactivity and better signal-to- noise ratio in FA can be achieved when the NF is set at least to 1.0., P. Slezák, I. Waczulíková, P. Bališ, A. Púzserová., and Obsahuje bibliografii
Paraoxonase 1 (PON1) seems to have a relevant role in detoxifying processes and in atherosclerosis. The aim of this study was to determine PON1 activity, the total antioxidant capacity, as well as entire lipid profile in children for screening of possible risk of atherosclerosis development. Serum PON1 arylesterase/paraoxonase activities were determined spectrophotometrically. The total antioxidant capacity of the serum was measured by TEAC method. Parameters of lipid profile were analyzed by routine laboratory methods. It has been shown that PON1 arylesterase/ paraoxonase activities were very similar to values found in adults. In children, no significant correlation between PON1 arylesterase activity and HDL was observed. PON1 paraoxonase activity correlated only with atherogenic index. PON1 arylesterase activity was significantly higher in girls than in boys. The antioxidant capacity was inversely related to the body mass index. In this study, PON1 activity was determined in healthy children aged 11 to 12 years and we found a similarity in PON1 activities of children and adults. Moreover, the results of our study support the hypothesis that higher body weight of children may contribute to a greater risk for development of atherosclerosis in which oxidative stress plays a role., K. Sumegová, Z. Nagyová, I. Waczulíková, I. Žitňanová, Z. Ďuračková., and Obsahuje bibliografii a bibliografické odkazy
Diabetes is a recognized risk factor of heart disease. The abnormalities related to a decreased heart performance probably arise at cellular and molecular levels already in the asymptomatic phase of diabetes. However, the early alterations initiating a sequence of events that culminates in the clinical signs have not been fully elucidated yet. This review deals with some biophysical methods applied to investigation of left ventricular myocytes in rats with streptozotocin diabetes, as well as our most important findings concerning diabetes-induced cell changes which cannot be captured by other techniques. The observed decrease in sarcolemmal membrane fluidity is causatively associated with increased glycation and glycoxidation. On the other hand, an increase in the mitochondrial membrane fluidity may be attributed to augmented energy transduction through the membranes. We reported for the first time concurrent measurements of membrane potential and dynamics, and respiratory chain activities in rat heart mitochondria, as well as calcium transients in the myocytes from diabetic hearts together with the assessed quantitative relationships among these variables. We were able to detect some significant alterations that may underlie myocyte dysfunction and subsequent remodeling of the heart. We suppose that not all these changes reflect mechanisms leading to pathology; some may represent adaptive and compensatory responses to diabetes., I. Waczulíková ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
A 2×2 factorial design was used to evaluate possible preservation
of mitochondrial functions in two cardioprotective experimental models, remote ischemic preconditioning and streptozotocin-induced diabetes mellitus, and their interaction during ischemia/reperfusion injury (I/R) of the heart. Male Wistar rats were randomly allocated into four groups: control (C), streptozotocin-induced diabetic (DM), preconditioned
(RPC) and preconditioned streptozotocin-induced diabetic (DM+RPC).
RPC was conducted by 3 cycles of 5-min hind-limb ischemia and 5-min reperfusion. DM was induced by a single dose of 65mg/kg streptozotocin. Isolated hearts were exposed to ischemia/reperfusion test according to Langendorff. Thereafter mitochondria were isolated and the mitochondrial respiration was measured. Additionally, the ATP synthase activity measurements on the same preparations were done. Animals of all groups subjected to I/Rexhibited a decreased state 3 respiration with the least change noted in DM+RPC group associated with no significant changes in state 2 respiration. In RPC, DM and DM+RPC group, no significant
changes in the activity of ATP synthase were observed after I/R
injury. These results suggest that the endogenous protective mechanisms of RPC and DM do preserve the mitochondrial function in heart when they act in combination.
The aim of present study was to investigate functional and physical alterations in membranes of heart mitochondria that are associated with remodeling of these organelles in acute phase of streptozotocin-induced diabetes and to elucidate the role of these changes in adaptation of the heart to acute streptozotocin-induced diabetes (evaluated 8 days after single dose streptozotocin application to male Wistar rats). Action of free radicals on the respiratory chain of diabetic-heart mitochondria was manifested by 17 % increase (p<0.05) in oxidized form of the coenzyme Q10 and resulted in a decrease of states S3 and S4 respiration, the respiratory control index, rate of phosphorylation (all p<0.01) and the mitochondrial transmembrane potential (p<0.05), but the ADP/O ratio decreased only moderately (p>0.05). On the contrary, membrane fluidity and the total mitochondrial Mg2+-ATPase activity increased (both p<0.05). In diabetic heart mitochondria, linear regression analysis revealed a reciprocal relationship between the increase in membrane fluidity and decrease in trans-membrane potential (p<0.05, r = 0.67). Changes in membrane fluidity, transmembrane potential, Mg2+-ATPase activity and the almost preserved ADP/O ratio appear as the manifestation of endogenous protective mechanisms participating in the functional remodeling of mitochondria which contributes to adaptation of the heart to diabetes., M. Ferko, D. Habodászová, I. Waczulíková, J. Mujkošová, J. Kucharská, L. Šikurová, B. Ziegelhöffer, J. Styk, A. Ziegelhöffer., and Obsahuje bibliografii a bibliografické odkazy
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
Statins, inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, are effective drugs in the treatment of hypercholesterolemia, however, their undesirable actions are not fully known. We investigated the effects of atorvastatin on the oxidative phosphorylation and membrane fluidity in liver mitochondria, and also on the coenzyme Q (CoQ) content in the mitochondria, liver tissue, and plasma of rats on a standard (C) and hypercholesterolemic (HCh) diet. Atorvastatin was administered at either low (10 mg⋅kg-1) or high dose (80 mg⋅kg-1) for four weeks. The high dose of the drug decreased the concentrations of total cholesterol and triacylglycerols in the plasma and liver of rats on a HCh diet. Administration of atorvastatin was associated with decreased oxygen uptake (state 3), and oxidative phosphorylation rate in the mitochondria of both C and HCh rats. Further, the drug influenced mitochondrial membrane fluidity and dose-dependently reduced concentrations of oxidized and reduced forms of CoQ in the mitochondria. Our findings point to an association between in vivo administration of atorvastatin and impaired bioenergetics in the liver mitochondria of rats, regardless of diet, in conjunction with simultaneous depletion of oxidized and reduced CoQ forms from the mitochondria. This fact may play a significant role in the development of statin-induced hepatopathy., O. Uličná ...[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