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PubMed1985

Effects of long-chain cis-unsaturated fatty acids and their alcohol analogs on aggregation of bovine platelets and their relation with membrane fluidity change.

The effects of long-chain cis-unsaturated fatty acids with different alkyl chain lengths and different numbers of double bonds on aggregation of bovine platelets and membrane fluidity were investigated. All the cis-unsaturated fatty acids tested inhibited aggregation and at the same time increased membrane fluidity in accordance with their inhibitory effects. The saturated fatty acids and trans-unsaturated fatty acid tested for comparison had much lower or no effects on aggregation and membrane fluidity. The inhibitory effects of mono cis-unsaturated fatty acids increased with increase of their alkyl chain length. cis-Unsaturated fatty acids with two or more double bonds had more inhibitory effects than mono-unsaturated fatty acids. The position of the double bonds had less influence than the number of double bonds. We also examined the effects of cis-unsaturated fatty acids on membrane fluidity with diphenylhexatriene and anthroyloxy derivatives of fatty acids as probes and observed increased fluidity to be considerable in the membrane. The alcohol analogs of cis-unsaturated fatty acids also inhibited aggregation and increased membrane perturbation. These results suggest that the inhibition of platelet aggregation by cis-unsaturated compounds is due to perturbation of the lipid layer.

Adenosine Diphosphate↗
PubMedDate not supplied

Plasma membrane fluidity and polarity of polymorphonuclear leukocytes from children with type I diabetes mellitus.

Polymorphonuclear leukocytes (PMN) from diabetic subjects have been found to be abnormal in various functional activities. These activities are mediated by the plasma membrane. This study was designed to evaluate plasma membrane fluidity and polarity in children with type I diabetes mellitus using fluorescence spectroscopy. PMN membrane fluidity and polarity were assessed in a group of 32 diabetic children. Membrane fluidity was investigated by measuring steady-state fluorescence anisotropy and fluorescence decay of 1-[4-trimethylammonium-phenyl]-6-phenyl- 1,3,5-hexatriene (TMA-DPH), whereas membrane polarity was studied by measuring the steady-state fluorescence emission and excitation spectra of 2-dimethylamino[6-lauroyl]-naphthalene (Laurdan). TMA-DPH and Laurdan are known to be incorporated at the hydrophobic-hydrophilic interface of the bilayer. Our data show a significant increase in steady-state fluorescence anisotropy in diabetic PMN that reflects a decrease in membrane fluidity, and a decrease in TMA-DPH lifetime distribution indicating a decrease in membrane heterogeneity. Laurdan shows a blue shift of the fluorescence emission and a red shift of the excitation spectra in diabetic PMN with respect to the control group, indicating a decrease in membrane polarity. The results demonstrate a decrease in the phospholipid order at the membrane surface and a decrease in membrane polarity in diabetic PMN. These alterations in the physico-chemical properties of the plasma membrane could be the basis of the modifications in functional activities of PMN. The changes in the plasma membrane of PMN could be the result of metabolic and chemical modification associated with type I diabetes.

Adolescent↗
PubMed1987

Influence of polyunsaturated fatty acid supplementation and membrane fluidity on ozone and nitrogen dioxide sensitivity of rat alveolar macrophages.

The phospholipid polyunsaturated fatty acid (PUFA) content and the membrane fluidity of rat alveolar macrophages were modified dose-dependently and in different ways. This was done to study the importance of both membrane characteristics for the cellular sensitivity toward ozone and nitrogen dioxide. Cells preincubated with arachidonic acid (20:4) complexed to bovine serum albumin (BSA) demonstrated an increased in vitro sensitivity versus ozone and nitrogen dioxide. The phenomenon was only observed at the highest 20:4 concentrations tested, whereas the membrane fluidity of the 20:4-treated cells already showed a maximum increase at lower preincubation concentrations. Hence it could be concluded that the increased ozone and nitrogen dioxide sensitivity of PUFA-enriched cells is not caused by their increased membrane fluidity, resulting in an increased accessibility of sensitive cellular fatty acid moieties or amino acid residues. This conclusion receives further support from other observations. These results strongly support the involvement of lipid oxidation in the mechanism(s) of toxic action of both ozone and nitrogen dioxide in an intact cell system.

Animals↗
PubMed2005

A nuclear receptor-mediated choleretic action of fibrates is associated with enhanced canalicular membrane fluidity and transporter activity mediating bile acid-independent bile secretion.

Fibrates are commonly used lipid-lowering agents that act via PPARalpha, a member of the nuclear hormone receptor superfamily. The mechanism(s) of fibrate-induced changes in the hepatic canalicular membrane and bile lipids are still unknown. Therefore, the aim of this study was to investigate the influence of fibrates on hepatic lipid metabolism and to assess the hepatocellular cytoprotective effect on hepatocyte canalicular membrane. Male ICR mice were fed standard chow with or without bezafibrate (100 mg/kg) for 6 days. The expression of canalicular membrane transporters (Mdr2 and Mrp2) was evaluated by RT-PCR and Western blotting. Canalicular membrane fluidity was also investigated. Canalicular membrane fluidity was markedly increased by fibrates. The expression of mdr 2 and mrp2 mRNA and protein showed a significant increase in fibrate-treated mice. These results suggested that fibrates improve liver function by enhancing bile secretion. The mechanism of the choleretic action of fibrate therapy might involve the enhancement of bile acid-independent bile secretion, since increased expression of Mdr2 and Mrp2 was found in fibrate-treated animals. These changes were very likely mediated by PPARalpha, and the increase of canalicular membrane fluidity may have been partly associated with enhancement of this transporter activity.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed1989

Reversal of ethinylestradiol-induced cholestasis by epomediol in rat. The role of liver plasma-membrane fluidity.

Epomediol (EPO) is a synthetic terpenoid compound shown to be active in increasing bile flow and some enzymatic activities of liver plasma membranes in the rat. The possible effect of EPO treatment in the ethinyl-estradiol (EE) induced cholestasis in the rat was investigated by measuring the hepatic transport of sulfobromophthalein (BSP) (plasma clearance and biliary secretion) and bile flow. Liver plasma membrane fluidity was also determined by the steady state fluorescence polarization (P) of diphenylhexatriene (DPH). EE administration (5 mg/kg s.c. for 5 days) was followed by a significant, comparable reduction (P less than 0.001) in BSP plasma clearance and biliary excretion and in bile flow. Intraperitoneal administration of EPO (100 mg/kg) to EE-treated rats restored both parameters of BSP transport, as well as bile flow, to control values. Liver plasma membrane fluidity was markedly (P less than 0.01) decreased by EE administration with a concomitant reduction (P less than 0.01) in Na+/K+-ATPase activity. EPO administration significantly increased membrane fluidity to values higher either to cholestatic (P less than 0.05) or control (P less than 0.05) animals. On the contrary, EPO did not influence Na+/K+-ATPase activity in either EE-treated or control animals. These data indicate that EPO fully reverses the impairments of BSP transport and bile flow induced by EE, possibly by reversing the decrease in liver plasma membrane fluidity induced by the synthetic estrogen. On the contrary, the EE-mediated decrease in Na+/K+-ATPase activity was not reversed by EPO.

Animals↗
PubMed1996

Fatty acid modification of membrane fluidity in Chinese hamster ovary (TR715-19) cells.

Dietary saturated fatty acids, especially lauric (12:0), myristic (14:0) and palmitic (16:0) acids, which are hypercholesterolemic, influence cell membrane fatty acid composition and affect LDL receptor function. When membrane phospholipid fatty acids in Chinese hamster ovary cells, containing the human LDL receptor, were modified (Hannah J. S. et al., 1995 Metabolism 44, 1428-1434), LDL receptor function was affected, but correlations with DPH-determined membrane fluidity were weak. The role of fluidity in various membrane domains with respect to the LDL receptor is examined here. Membrane fluidity was assessed by measuring steady-state fluorescence polarization of diphenylhexatriene (DPH) and its polar propionic acid (DPH-PA) and trimethylammonium (TMA-DPH) derivatives from 38 to 4 degrees C in fatty acid modified Chinese hamster ovary cells. Fatty acid changes modulated mid-bilayer fluidity as determined with DPH, but fluidity in phospholipid headgroup domains, assessed with DPH-PA and TMA-DPH, was independent of fatty acyl composition. The DPH fluidity was related to membrane unsaturation (P < 0.02), oleate contents (P < 0.009) in particular, but inversely related (P < 0.0002) to the longer chain (> or = 20 C atoms) unsaturated fatty acids with from four to six double bonds. The LDL binding was independent of fluidity, but there were weak relations between LDL internalization and DPH-PA anisotropy and between LDL degradation and TMA-DPH anisotropy. It was concluded that LDL binding was not related to mid-bilayer fluidity, but the results with the polar probes suggest a role of fluidity in modulating vertical displacement of the LDL/LDL receptor complex across the plasma membrane.

Animals↗
PubMed2000

[The influence of epidermal growth factor, neurotensin on cytosolic calcium and membrane fluidity in carbon tetrachloride-injured primary cultured hepatocytes].

OBJECTIVE: To study the influence of epidermal growth factor (EGF), neurotensin (NT) on cytosolic calcium and membrane fluidity in carbon tetrachloride-injured primary cultured hepatocytes. METHODS: A micro-model of carbon tetrachloride -induced primary cultured hepatocytes injury was established. EGF and NT were added to the cells 1 h ahead of carbon tetrachloride given. Twenty-four hours later, cytosolic calcium and membrane fluidity in hepatocytes were detected with fluorescence probe of Fura-2/AM and 1, 6-dihenyl-1, 3, 5-hexatriene (DPH). RESULTS: EGF and NT could significantly antagonize the increase of cytosolic Ca(2+) and decrease of membrane fluidity in carbon tetrachloride-injured hepatocytes. CONCLUSION: EGF and NT have a function of maintaining membrane fluidity and steady-status of cytosolic calcium of hepatocytes.

Animals↗
PubMed1995

Cytosolic Ca2+ content and membrane fluidity of platelets and polymorphonuclear leucocytes in diabetes mellitus.

Considering the role played by platelets and leucocytes in diabetic disease and keeping in mind the strong correlation between functional and metabolic aspects that characterizes this clinical condition, we evaluated, in two groups of diabetics, respectively the platelet and polymorphonuclear (PMN) cytosolic Ca2+ content (employing the fluorescent probe Fura 2-AM) and membrane fluidity (using the fluorescent probe TMA-DPH and considering the fluorescence polarization degree, inversely related to the membrane fluidity). From the obtained results, it is evident that the platelet cytosolic Ca2+ content does not distinguish normals from diabetics of type 1 and 2; the platelet membrane fluidity instead does not discriminate normals from diabetics, but differentiates diabetics of type 1 and 2 (type 1 = 0.284 +/- 0.015; type 2 = 0.314 +/- 0.018; p < 0.001). PMN cytosolic Ca2+ content and membrane fluidity do not discriminate normals from diabetics. In the two groups of diabetics none of the platelet and PMN parameters (cytosolic Ca2+ content and membrane fluidity) are related to the glycometabolic pattern.

Adolescent↗
PubMed1997

Calmodulin-related changes in microsomal membrane fluidity during liver regeneration.

Based on our previous findings on the modifying effect of calmodulin (CaM) on the physiochemical properties of biomembrane, we have investigated the possible relationship between intracellular CaM content and endoplasmic reticulum (ER) membrane fluidity and function during liver regeneration. The degree of ER membrane fluidity was estimated by fluorescence polarization analysis with the 1,6-diphenyl-1,3,5-hexatriene probe. Microsomal guanylate cyclase (GC) was used as a functional parameter. The kinetics of the increase in the ER membrane fluidity during liver regeneration was strictly parallel to the CaM surge and was matched by an increase in GC activity. The stimulative effect of splenectomy on liver regeneration and its inhibition by Walker-256 tumor, inferred from the corresponding alterations of CaM levels, were mirrored by the modulation in GC activity. The fluidizing effect of CaM on ER membrane was concluded from the drop in thermotropic transition temperature from 28.3 +/- 1.6 degrees C in control membranes to 17.8 +/- 1.1 degrees C membranes from regenerating livers and to 19.8 +/- 1.2 degrees C in control membranes treated with CaM. Arrhenius plots of GC activity exhibited a transition temperature of 25.5 +/- 1.25 degrees C in controls, which shifted to 20.5 +/- 0.9 degrees C in ER membranes from regenerating livers and to 21.7 +/- 1.1 degrees C in control membranes treated with CaM. The Hill coefficient for the allosteric activation of the GC by Mn.GTP decreased from 1.49 +/- 0.16 in controls to 0.93 +/- 0.085 in membranes from regenerating cells and to 0.86 +/- 0.073 in CaM-treated membranes. Both effects of CaM were consistent with a fluidity increase in the enzyme's lipid microenvironment. The results of the present study suggest that an early key event in liver regeneration may be the CaM-induced modulation of ER membrane fluidity and function.

Animals↗
PubMed1999

Association of age-related decrease in platelet membrane fluidity with platelet lipid peroxide.

The influence of age on platelet lipid peroxide (LPO), platelet membrane fluidity and the composition of fatty acid was investigated in female Wistar rats widely ranging in age from 14 to 720 days old. LPO levels were significantly higher (p<0.05) in the platelets of upper age groups than in those of lower age groups, showing a significantly positive correlation with age (r=0.84, p<0.0001). Membrane fluidity, assessed by 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence polarization, was significantly reduced with age. The composition of fatty acid demonstrated an age-related elevation (p<0.05) in the unsaturation index. The rises in the LPO levels revealed a significantly positive correlation with DPH-polarization (r=0.73, p<0.0001). Thus our results suggested that the age-related deterioration of platelet membrane fluidity, despite a significant elevation in the unsaturation index, was due to the age-related higher basal levels of LPO in platelets.

Aging↗
PubMed1991

[Changes in the plasma membrane fluidity of granulosa cells during the process of luteinization].

To elucidate the membrane fluidity of granulosa cells in luteinization process, porcine granulosa cells (GCs) were cultured in the serum free medium under the stimulation of human chorionic gonadotropin (hCG). Their cell membrane anisotropy, membrane lipid composition and progesterone synthesis were measured. The GCs plasma membrane anisotropy, which is reverse to fluidity, was measured with the steady-state spectrofluorometer using a hydrophobic fluorescent probe 1, 6-diphenyl-1, 3, 5-hexatriene (DPH). Since the membrane fluidity is generally affected by concentrations of cholesterol and phospholipids, phospholipid composition and the degree of unsaturation of acyl chain, concentrations of unesterified cholesterol and phospholipids, and phospholipid composition in the plasma membranes of GCs were determined. The results obtained were as follows. 1) The value of the anisotropy (membrane fluidity) of the GCs stimulated by hCG decreased (increased) significantly as compared to that without hCG addition. 2) The concentration of cholesterol in plasma membrane fraction decreased significantly by hCG stimulation. 3) In contrast, the concentration and composition of phospholipids showed no significant changes. 4) Progesterone concentration in culture medium was significantly increased by hCG stimulation at 30 mlU/ml and 300 mlU/ml throughout the time course of this study, compared with control. These results indicate the increase of membrane fluidity of the GCs luteinization induced by hCG is due to a decrease of cholesterol/phospholipids ratio. Because transfer of cholesterol into culture medium was not observed, it seems that the cholesterol of the GCs plasma membranes was utilized for progesterone synthesis at least in the serum free medium (GIT medium) during the process of luteinization.

Animals↗
PubMed2003

Changes in plasma membrane fluidity lower the sensitivity of S. cerevisiae to killer toxin K1.

The possible correlation between plasma membrane fluidity changes induced by modified cultivation conditions and cell sensitivity to the killer toxin K1 of Saccharomyces cerevisiae were investigated. Cells grown under standard conditions exhibited high toxin sensitivity. Both a membrane fluidity drop and fluidity rise brought about markedly reduced sensitivity to the toxin. These results do not fit the hypothesis of physiological relevance of direct toxin-lipid interaction, suggesting that the essential event in killer toxin action is interaction with membrane protein(s) that can be negatively influenced by any changes of membrane fluidity.

Anti-Infective Agents, Local↗
PubMed1979

The decreased membrane fluidity of in vivo aged, human erythrocytes. A spin label study.

The decreased membrane fluidity of the in vivo aged, human erythrocytes is found, by monitoring the electron paramagnetic resonance (EPR) spectra of fatty acid spin labels incorporated into the membrane. In addition, the decreased cell sizes and the decreased cholesterol and phospholipids contents, without significant changes of the quantity of the membrane proteins, also the decrease of ATP and 2,3-diphosphoglycerate and the increase of ADP and AMP, in the aged cells, were observed. Further the functional impairments of the aged cells, i.e. the increased oxygen affinity and the decreased deformability, were shown. On the basis of these quantitative data, the alteration of the protein-lipid organization, due to decreased lipid/protein ratio, the modified protein-lipid interaction and/or the influences of the diminished ATP content, is suggested to contribute towards the decreased membrane fluidity of the in vivo aged erythrocytes.

Adenine Nucleotides↗
PubMed2003

Choleretic action of diosgenin is based upon the increases in canalicular membrane fluidity and transporter activity mediating bile acid independent bile flow.

It is known that diosgenin alters the lipid composition of hepatic plasma membranes as well as bile lipids. We recently reported that changes in the lipid composition of the canalicular membrane bilayer were associated with alterations of membrane fluidity. Therefore, the present study was performed to determine the effect of diosgenin on bile secretion, focusing on canalicular membrane composition, membrane fluidity, transporter expression, and transporter activity. METHODS: SD rats were fed a diet with or without diosgenin. Bile duct cannulation was done and the bile acid pool was depleted, followed by infusion of sodium taurocholate for 120 min to collect bile samples. Bile flow and the biliary output of cholesterol (Ch), phospholipids, bile salt, and total glutathione (GSH+GSSG) were estimated. Liver canalicular membrane vesicles (CMV) were prepared for the assessment of lipid composition, ATP-dependent transporters, and membrane fluidity. RESULTS: Bile flow, especially bile acid-independent fraction, was increased significantly by diosgenin. Biliary output of Ch and total glutathione were significantly increased by diosgenin. CMV fluidity and Mrp2 expression were increased by diosgenin. CONCLUSION: The fact that diosgenin increased bile flow and biliary cholesterol and glutathione output indicates a choleretic action along with enhancement of solute secretion. The increase of CMV fluidity and Mrp2 suggests that the choleretic action of diosgenin was based upon both the direct stimulation of transporter expression and indirect transporter activation by an increase of membrane fluidity.

Journal Article↗
PubMed1991

[Studies on correlation between liver drug metabolizing enzyme activities and microsomal membrane fluidity in phenobarbital treated rats].

By the use of ANS(1-anilinonaphthalene-8-sulfonate) and DPH(1, 6-diphenyl-1, 3, 5-hexatriene) as fluorescent probes, correlation between liver microsomal membrane fluidity and drug metabolizing enzyme activity has been studied in rats. phenobarbital(PB) ip treatment caused an increase in P-450 content, cytochrome C reductase.aminopyrine N-demethylase(AMD) and glutathione S-transferase(GST) activities by 78, 66, 270 and 52%, respectively. However, there was a simultaneous decrease in microsomal membrane fluorescent intensity and microviscosity, i.e. an increase in membrane fluidity. There is a positive linear correlation between microsomal membrane fluidity and cytochrome C reductase and AMD activities (r = 0.798, r = 0.781, respectively, P less than 0.05). This result suggests that there may be some relationship between microsomal membrane fluidity and drug-metabolizing enzymatic activities in PB-treated rats.

Aminopyrine N-Demethylase↗
PubMed1999

Impairment of the erythrocyte membrane fluidity in survivors of acute myocardial infarction. A prospective study.

Erythrocytes have to constantly adapt themselves to the varying circulatory system shear stress forces and capillaries diameter. Membrane lipid and protein content have an important role in determining the erythrocyte shape and are main determinants of the membrane solid and fluid behaviour which enables the erythrocyte to respond to the outer environment modifications. Membrane fluidity is an inverse index of membrane microviscosity. The aim of the present work is to evaluate prospectively in three periods of time (discharge, after 6 months and one year later) in survivors of an acute myocardial infarction (AMI) the erythrocyte membrane fluidity (outer and inner bilayer) and establish a relation with the cardiovascular events or need of coronary revascularization during a two year clinical follow up. Sixty survivors of acute myocardial infarction were recruited during 1994-96 and were prospectively studied in three periods (discharge, 6 months and after one year), and were compared with a control group (n = 36). Membrane lipid fluidity was determined by means of fluorescence polarisation with two probes: 1,6-diphenyl-1,2,5-hexatriene (DPH) and 1,4-trimethylamine 6-phenyl hexa-1,3,5-triene (TMA-DPH), for the characterisation of the hydrophobic and external polar region, respectively. The hydrophobic region was more rigidified (p < 0.01) in the erythrocytes from AMI patients, in relation to the control group. During the time of the study there was a progressive erythrocyte membrane rigidification (DPH p < 0.001; TMA-DPH p < 0.001). We found no relation between erythrocyte membrane fluidity and the coronary risk factors, cardiovascular events or the need of coronary revascularization during the clinical follow-up. In conclusion, after the myocardial infarction erythrocyte membrane of AMI survivors becomes more rigid with time, which could contribute to the decreased erythrocyte deformability and the increased blood viscosity previously described in this group of patients.

Aged↗
PubMed1983

Cholesteryl hemisuccinate alters membrane fluidity, angiotensin receptors, and responses in adrenal glomerulosa cells.

We examined the effects of cholesteryl hemisuccinate on membrane fluidity and angiotensin II (AII) actions in bovine adrenal glomerulosa cells. Incubating cells with cholesteryl hemisuccinate decreased membrane fluidity and markedly inhibited AII binding. The effect on binding was characterized by a decrease in AII receptor number. The effects of AII on phosphatidyl inositol turnover and calcium fluxes, proposed intermediaries of AII actions on aldosterone secretion, were less impaired than AII binding by cholesteryl hemisccinate. AII stimulation of aldosterone secretion was preserved despite the decrease in AII binding after cholesteryl hemisuccinate treatment. These results indicate that AII binding can be dissociated from its effects on aldosteronogenesis by a reagent that alters membrane fluidity.

Adrenal Cortex↗
PubMed2006

Decreased protein tyrosine phosphorylation and membrane fluidity in spermatozoa from infertile men with varicocele.

Varicocele is a prevalent pathology among infertile men. The mechanisms linking this condition to infertility, however, are poorly understood. Our previous work showed a relationship between sperm functional quality and the ability of spermatozoa to respond to capacitating conditions with increased membrane fluidity and protein tyrosine phosphorylation. Given the reported association between varicocele, oxidative stress, and sperm dysfunction, we hypothesized that spermatozoa from infertile patients with varicocele might have a combined defect at the level of membrane fluidity and protein tyrosine phosphorylation. Semen samples from infertile patients with and without grade II/III left varicocele were evaluated for motion parameters (computer-assisted semen analysis [CASA]), hyperactivation (CASA), incidence and intensity of protein tyrosine phosphorylation (phosphotyrosine immunofluorescence and western blotting), and membrane fluidity (Laurdan fluorometry), before and after a capacitating incubation (6 hr at 37 degrees C in Ham's F10/BSA, 5% CO(2)). Spermatozoa from varicocele samples presented a decreased response to the capacitating challenge, showing significantly lower motility, hyperactivation, incidence and intensity of tyrosine phosphorylation, and membrane fluidity. The findings reported in this article indicate that the sperm dysfunction associated to infertile varicocele coexists with decreased sperm plasma membrane fluidity and tyrosine phosphorylation. These deficiencies represent potential new pathophysiological mechanisms underlying varicocele-related infertility.

Adult↗