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1.
The aim of the present work was to study by ESR-spin-probe technique the effect of the natural antioxidant alpha-tocopherol (alpha-tp) in vitro on the structural parameters (microviscosity, order parameter) of endoplasmic reticulum membranes of the mice liver cells starting from the concentration of 10(-3) mol/l and down to the dilution of 10(-25) mol/l. The stable nitroxyl radicals 16-doxylstearic acid (with the deep localization depth of 20 A) and 5-doxylstearic acid (with the surface localization depth of 8 A) were used as spin probes. It has been shown that alpha-tp causes the increase in microviscosity of the deep lipid bilayer regions and in rigidity of the surface ones at the certain concentrations. The concentration curves obtained have the polymodal shapes being typical of the effects of substances at ultra low doses. Using 16-doxylstearic acid it is detected the increase in the number of thermoinduced structural transitions and appearance of much more high-cooperative ones, as well as the increase in their effective activation energy with the rise of temperature at the supplement of different alpha-tp doses.  相似文献   

2.
The effect of the natural antioxidant alpha-tocopherol in a broad concentration range (10(-4) - 10(-25) M) on the viscosity characteristics and thermally induced structural transitions of a lipid bilayer of plasma membranes of murine hepatocytes in vitro has been studied. Changes in the rigidity of surface (approximately Abb) of the lipid bilayer were measured on a Bruker EMX EPR spectrometer (Germany) by the method of spin probes. Stable nitroxyl radicals of 5- and 16-doxylstearic acid, localized at different depth in the membrane served as spin probes. It was shown that the concentration dependence of the effect of alpha-tocopherol is linear and polymodal with three statistically significant increases in three ranges of its concentration: (1) in the range of traditional physiological concentrations 10(-4)-10(-9) M, (2) in the range of superlow doses 10(-9) - 10(-17) M, and (3) in the range of "imaginary" concentrations 10(-17) - 10(-25) M. The mechanisms of action of alpha-tocopherol in each of the three ranges are discussed. When studying the temperature dependences of viscous characteristics, a new thermally induced structural transition in the range of "physiological" temperatures 309-313 K for those alpha-tocopherol concentrations (including superlow ones) to which the maxima on the dose dependence curves at constant temperature of 293 K corresponded.  相似文献   

3.
The influence of tocopherol and its analogue (oxychroman) on the microviscosity of mitochondrial lipids was studied, using spin labels. The viscosity of the lipid bilayer was shown to enhance with the increase in the antioxidant content in the membrane. Small concentrations of alpha-tocopherol (10(-5)-10(-6) mol/l) were shown to increase, while large concentrations (10(-3)-10(-4) mol/l) decreased the fluidity of the lipid bilayer. The influence of alpha-tocopherol on fluidity of the lipid bilayer depending on its concentration could be realized in two ways: by direct influence on the lipid bilayer and via reception. It was shown that alterations in the viscosity of the lipid bilayer depend on chroman cycle of tocopherols, while the temperature of structural transfer and effective energy of activation depend on the lateral phytyl chain.  相似文献   

4.

In this study, the effects of different concentrations of antioxidant resveratrol on microviscosity of mitochondrial membranes isolated from pea leaves were investigated. It was shown that resveratrol in physiological (5 × 10–6 M) and ultralow (5 × 10–14 M) doses changed the microviscosity and altered the structure of the membrane lipid bilayer. This antioxidant is thought to affect the activity of membrane proteins due to altered structures of lipid molecules that surround them and, in turn, influence mitochondrial activity and function. No change was found in the range between physiological and ultralow doses, which is a “dead” zone. Moreover, resveratrol in physiological concentrations was found to shift thermally induced structural transition to the lower temperature region; therefore, it can interfere with the normal regulation of natural processes

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5.
Peptides are known to have the ability of modulating the activity of important regulatory cellular systems. One of them--thyroliberin, i.e. thyreotropin-releasing hormone (TRH), causes changes in the membrane structure and morphology of rat erythrocytes, as well as activates retractive activity of lymphatic vessels in ultra low concentrations (10(-10) to 10(-16) mol/l). In this study we used an electron spin resonance (ESR) method to explore the effect of TRH in a wide range of concentrations (10(-4) to 10(-18) mol/l) on thermo-induced structural transitions and microviscosity of lipid bilayer of the endoplasmic reticulum membrane of mice (C57 bI) liver cells. Two stable free radicals were used as paramagnetic probes: 2,2,6,6-tetramethil-4-capryolyl-1-oxyl and 16-doxyl-stearic acid, that are localized in superficial and deep layers of the membrane respectively. TRH caused a statistically significant change (p < 0.001) in microviscosity of the membrane surface layer. The largest effect (up to 30% decrease) was observed at TRH concentrations of 10(-10) and 10(-16) mol/l. It was also demonstrated that an addition of 10(-4), 10(-10) and 10(-16) mol/l of TRH decreases effective activation energy and temperature (by several degrees) of the thermo-induced structural transitions. The observed changes in the parameters of the membrane surface layer induced by TRH may be essential for its physiological activity, because of the obtained negative correlation (r = 0.99; p < 0.001) between the membrane microviscosity and frequency of lymphatic vessels' contraction. Complex changes in the structure of deep hydrophobic layer of the membrane caused by TRH were observed in this study as well. Higher concentrations of TRH (10(-4) and 10(-10) mol/l) produced results that were similar to the effect of TRH on the superficial lipid layer of the membrane, whereas the effect of ultra low TRH concentration (10(-16) mol/l) was reversed for microviscosity, number and activation energy of structural transitions in contrast with the case of surface layer. The results of this study suggest presence of a nonspecific factor in the effect of TRH on structural characteristics of the lipid component of biological membranes. It is possible, that the change of structural properties of biological membranes may be a part of the mechanism of TRH action at ultra low concentrations.  相似文献   

6.
The effect of lead acetate on the physical state of membrane lipids in human erythrocytes in vitro was studied using the lipophilic fluorescence probe 1,6-diphenyl-1,3,5-hexatriene and spin probes 16-doxyl-stearate and iminoxyl palmitic acid. It was shown that 2-10 microM lead acetate causes an increase in both intensity and polarization of fluorescence of 1,6-diphenyl-1,3,5-hexatriene, indicating changes in the microviscosity of the lipid bilayer of erythrocyte membranes. Judging from the parameters of EPR spectra of 16-doxyl stearate and iminoxyl palmitic acid incorporated into erythrocyte membranes, 2-10 microM lead acetate increases the heterogeneity of the lipid bilayer in surface and deep hydrophobic layers of the erythrocyte membrane.  相似文献   

7.
Using fluorescent and EPR spin probing techniques, the effects of phospholipases A2, C and D on rat brain synaptosomal membranes were investigated. It was shown that treatment of synaptosomal membranes with phospholipases A2, C and D results in their depolarization and increase of their surface negative charge. In case of phospholipases A2 and C, these changes are also accompanied by a decrease of the microviscosity of the synaptosomal membrane lipid bilayer. alpha-Tocopherol protects synaptosomal membranes against the damaging action of phospholipases. The stabilization of synaptosomes by vitamin E consists in the reconstitution of the transmembrane potential and in an increased microviscosity of phospholipase-treated membranes. The stabilizing effect of alpha-tocopherol is due to the binding of phospholipid hydrolysis products rather than to the inhibition of phospholipases. The observed stabilization of synaptosomal membranes by alpha-tocopherol is interpreted as a feasible mechanism of biological effects of vitamin E on biological membranes.  相似文献   

8.
Mitochondria were isolated from sugar beet (Beta vulgaris L) taproots and incubated in the presence of low concentrations of Melafen (2 × 10?9 and 4 × 10?12 M). This treatment of mitochondrial membranes induced an appreciable decrease in microviscosity of superficial lipids in the lipid bilayer and a parallel increase in microviscosity of the deeply immersed lipid regions adjacent to membrane proteins. Melafen had no effect on fluorescence of lipid peroxidation products in membranes of freshly prepared mitochondria but declined this fluorescence to control values in artificially aged mitochondria. Melafen raised the maximum rates for oxidation of NAD-dependent substrates, elevated the efficiency of oxidative phosphorylation, and activated electron transport in the terminal (cytochrome oxidase) step of mitochondrial respiratory chain, which implies the activation of energy metabolism within the cell. The acceleration of electron transport through the terminal step of mitochondrial respiratory chain was apparently accompanied by retardation of lipid peroxidation, which prevented impairment of mitochondrial membranes under stress conditions. A proposal is put forward that some properties of Melafen are favorable for adaptogenesis because its effects on mitochondrial energy metabolism depended on the functional state of mitochondria.  相似文献   

9.
The effect of synthetic anti-oxidant potassium phenosan (PP, potassium salt of β-(4-hydroxy-3,5-ditretbutil-phenyl)-propionic acid) on the structural state of the surface (8 Å) and deep (20–22 Å) lipid regions of plasma membranes of mice liver cells was studied by spin probes method in vitro in a wide range of concentrations (10?5–10?21 M). Two stable free radicals, 5- and 16-doxyl-stearic acids (C5 and C16), were used as spin probes. The nonlinear polymodal dose-effect dependences were obtained for parameters that characterize the microviscosity of the lipid bilayer (τc) in the site of localization of the probe C16, and the order parameter (S), which characterizes the stiffness of the surface layers of lipids in the site of localization of the probe C5. Statistically a reliable increase was observed for parameter τc after addition of PP at concentrations 10?5–10?7 M and 10?18–10?19 M, and for parameter S after addition of PP at concentrations 10?6–10?7 M and 10?13–10?15 M. Peaks on both dose-effect curves were separated by the intervals of concentrations where PP had no effect on the studied physico-chemical characteristics of biomembranes. For PP concentrations which caused maximal changes in τc and S, we investigated thermal dependence of these parameters and determined the thermally induced structural transitions. Comparing with control, ultra-low doses of PP (10?13–10?15 M) and (10?18–10?19 M) caused an appearance of additional thermally induced structural transition in the surface and deep regions of plasma membrane lipids. The possible role of the interaction of PP molecules with specific binding sites on plasma membranes and formation of nanoparticles of PP in very dilute aqueous solutions are discussed.  相似文献   

10.
Interaction of alpha-tocopherol and 1,4-dihydropyridine with endoplasmic reticulum membranes and model systems, human serum albumin and phospholipid bilayer, was studied using the microcalorimetry and fluorescent probes procedures. Dependence of microviscosity changes in the endoplasmic reticulum membranes on the place of antioxidants localization (protein structures or phospholipid phase) was shown. Increase of membrane structuralization under the influence of 1,4-dihydropyridines blocked their antioxidant action in spontaneous and induced lipid peroxidation.  相似文献   

11.
H Hauser  N Gains  G Semenza  M Spiess 《Biochemistry》1982,21(22):5621-5628
The temperature dependence of the packing (order) and fluidity (microviscosity) of rabbit small, intestinal brush border vesicle membranes and of liposomes made from their extracted lipids has been investigated by using a variety of lipid spin probes. The lipids in the brush border membrane are present essentially as a bilayer. Compared to other mammalian membranes, the brush border membrane appears to be characterized by a relatively high packing order as well as microviscosity. At body temperature, the lipid molecules undergo rapid, anisotropic motion, which is essentially a fast rotation about an axis approximately perpendicular to the bilayer normal. Both the order (motional anisotropy) and the microviscosity increase with decreasing temperature and with increasing distance from the center of the bilayer. Qualitatively similar motional or fluidity gradients have been reported for other mammalian and bacterial membranes. The liposomes made from the extracted lipids have a somewhat lower packing order and a slightly higher fluidity than brush border vesicle membranes. The differences are, however, small indicating that the packing and the fluidity (microviscosity) of the membrane are primarily determined by the lipid composition. Membrane-associated proteins and cytoskeleton cannot play a dominant role in determining the order and fluidity of the lipid bilayer. Discontinuities are observed in the temperature dependence of various spectral parameters, the order parameter S, the rotational correlation time tau, and 2,2,6,6-tetramethylpiperidinyloxy partitioning. They are assigned to phase transitions and/or phase separations of the membrane lipids. These discontinuities occur at about 30, 20, and 13 degrees C for 5-doxyl-, 12-doxyl-, and 16-doxylstearic acid, respectively. The apparent transition temperature depends on the location of the spin probe along the bilayer normal, being higher the closer the probe is to the membrane surface. This indicates the possibility that chain melting is progressive and spreads with increasing temperature from the center of the membrane outward.  相似文献   

12.
Physico-chemical parameters of membranes of skeletal muscles' sarcoplasmic reticulum in antioxidant insufficiency, which was modelled by excluding alpha-tocopherol from the animals ration, and after treatment with phenol antioxidant ionol were studied. It was shown that activation of lipid peroxidation in vitamin E insufficiency results in a significant lowering of microviscosity of lipid bilayer membranes of sarcoplasmic reticulum. Using polarography significant changes in membrane protein conformation were revealed, which were characterized by lowering of integrity and by disorganization of protein globules. Treatment of animals with antioxidant insufficiency with ionol led to certain normalization of changes of physico-chemical characteristics of the learned membrane structures caused by lipid peroxidation.  相似文献   

13.
Cell plasma membranes and proteoliposomes reconstituted from solubilized plasma membranes of thymocytes and Ehrlich ascites carcinoma cells have been studied by fluorescent methods. It has been shown that proteoliposomes are characterized by greater polarization and rigidity of microsurroundings in membrane proteins and greater microviscosity of membrane lipids. Proteoliposomes from thymocyte membranes contain less membrane proteins and express lower polarity of the lipid bilayer than proteoliposomes from Ehrlich ascites cells.  相似文献   

14.
The action of 12-O-tetradecanoyl-13-acetate (TPA) in vitro in a wide range of concentration from 10(-3) mol/l down to ultra-low doses 10(-23) mol/l and dilution 10(-24) mol/l on the microsome membranes isolated from tumor--Ehrlich ascite carcinoma (EAC) has been studied by ESR-method using two spin probes: 5- and 16-doxyl stearates (5- and 16-DS) localized in the different regions of lipid bilayer. From the ESR spectra obtained it was calculated the following parameters: an order of the long axis 5-DS (S) related to order of the fatty acids chains in the lipid bilayer; two rotation correlation times (Tc1 and Tc2) of 16-DC to estimate a microviscosity value and structural-sensitive ones. It was found the stage changes of all these parameters (increase and decrease) as compared with control level (the membranes untreated by TPA) depending on TPA concentration into the range of 10(-3)-10(-24) mol/l; in particular, the most significant shape changes of structural-sensitive parameters have been observed at TPA doses below 10(-16) mol/l. It is concluded that tumor membranes are very sensitive to TPA action in vitro in a wide range of concentration included ultra-low doses.  相似文献   

15.
The microviscosity of artificial lipid membranes and natural membranes was measured by the fluorescence polarization technique employing perylene as the probe. Lipid dispersions composed of brain gangliosides exhibited greater microviscosity than phosphatidylserine (268 cP vs 173 cP, at 25 degrees C). Incorporation of cholesterol (30-50%) increased the microviscosity of lipid phases by 200-500 cP. Cholesterol's effect on membrane fluidity was completely reversed by digitonin but not by amphotericin B. Incorporation of membrane proteins into lipid vesicles gave varying results. Cytochrome b5 did not alter membrane fluidity. However, myelin proteolipid produced an apparent increase in microviscosity, but this effect might be due to partitioning of perylene between lipid and protein binding sites since tha latter have a higher fluorescence anisotropy than the lipid. The local anesthetics tetracain and butacaine increased the fluidity of lipid dispersions, natural membranes and intact ascites tumor cell membranes. The effect of anesthetics appears to be due to an increased disordering of lipid structure. The fluidity of natural membranes at 25 degrees C varied as follows: polymorphonuclear leukocytes, 335 cP; bovine brain myelin, 270 cP; human erythrocyte, 180 cP; rat liver microsomes, 95 cP; rat liver mitochondria, 90 cP. In most cases the microviscosity of natural membranes reflects their cholesterol: phospholipid ratio. The natural variations in fluidity of cellular membranes probably reflect important functional requirements. Similarly, the effects of some drugs which alter membrane permeability may be the result of their effects on membrane fluidity.  相似文献   

16.
Fluorescent probes 1,6-diphenyl-1,3,5-hexatriene (DPH) and pyrene were employed in studying the effect of aminazine and triftazin versus that of imipramine on microviscosity of rat brain cortex synaptosomal membranes. Unlike imipramine, the neuroleptics decrease microviscosity of membrane's lipid bilayer. All drugs decrease fluorescence of endogenous tryptophan, but fail to change fluorescence of L-tryptophan in the solution. It is concluded that neuroleptics induce conformational perturbations in membrane-bound proteins modifying microviscosity of lipid bilayer whereas imipramine changes the surface electric charge of lipid bilayer of synaptosomal membranes.  相似文献   

17.
It was shown that low doses of phenozan (10(-17), 10(-14), 10(-11), 10(-5) mol/kg) caused changes in kinetic parameters Vmax, Km of LDG and microviscosity in various ranges of microsomal membranes of brain cells. It is suggested that the effect of phenozan on the structure of bilayer membrane is important for binding LDG with membranes.  相似文献   

18.
Using the high resolution 1H-NMR spectroscopy and spin-probes the influence of alpha-tocopherol on lipid bilayer microviscosity has been studied. It has been established that alpha-tocopherol shows the cholesterol-like action on the physical state of lipid bilayer: alpha-tocopherol increase microviscosity of unsaturated bilayers and decrease microviscosity of saturated bilayers. The character of alpha-tocopherol action is determined by the fatty acidic lipid composition but does not depend on the polar group structure of phospholipid molecule as cholesterol-like action of alpha-tocopherol is found itself in liposomes prepared both from phosphatidylcholine and phosphatidylethanolamine. Analog of alpha-tocopherol without phytol chain 2,2,5,7,8-penthamethyl-6-oxychroman does not show the cholesterol-like action as it is not able to disorder the saturated bilayers.  相似文献   

19.
Effects of low (from 4 × 10?12 to 2 × 10?7 M) doses of the organophosphorus plant growth regulator Melaphen on structural characteristics of plant and animal cellular membranes were compared with special reference to changes in the microviscosity of free membrane lipid bilayers and annular lipids bound to protein clusters. It was found that effective concentrations of Melaphen were not only different for animal and plant membranes, but also discrete and equal to 2 × 10?7 or 4 × 10?12 M depending on the membrane origin and the nature of membrane lipid components. In parallel experiments, effects of Melaphen on the rate of lipid peroxidation (LPO) in biological membranes were studied under conditions of external cold stress. The intensity of LPO was decreased at all Melaphen concentrations able to modulate the microviscosities of free and annular membrane lipids. It is concluded that effects of low and ultra-low Melaphen concentrations on structural and functional states of biological membranes of plant and animal origin are mediated by its interaction with signaling receptors of cellular membranes and cell organelles of both plant and animal origin.  相似文献   

20.
The degree of microviscosity, gh, (fluidity/rigidity behavior) of membrane lipids of normal and transformed mammalian fibroblasts obtained from mice, hamsters and rats was quantitatively monitored by fluorescence polarization, P, analysis of the fluorescent probe 1,6-diphenyl 1,3,5-hexatriene (DPH) when embedded in lipid regions of cellular membranes of intact viable cells. Analysis of membrane microviscosity of six different cell populations and of individual cells in each cell population have indicated that the membrane microviscosity of all cell types, both normal and transformed fibroblasts, changes as a function of the cell density in the growing cultures. The membrane microviscosity was found to be low (high lipid fluidity) in sparse conditions but high (high lipid rigidity) in dense conditions. The induced changes in membrane microviscosity are practically reversible for all cell types and a complete reversion can be obtained within a few hours after changing the cell density conditions from sparse to dense and vice versa.Comparative studies with normal and transformed fibroblasts have shown that transformed fibroblasts have a more rigid lipid layer in their cellular membranes than normal or untransformed fibroblasts. The difference in membrane microviscosity between transformed and normal fibroblasts is higher in confluent conditions as compared with subconfluent cultures. These differences in the degree of fluidity of membrane lipids that are controlled by possible differences in the cellto-cell contact in normal and transformed fibroblasts may play a major role in determining the growth behavior of normal and malignant cells that are growing as a solid tissue and may have a direct effect on the control mechanisms that determine the presence or absence of the “density dependent inhibition” of growth.  相似文献   

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