Decrease In Membrane Microviscosity Research Articles

Tinospora Cordifolia and Arabinogalactan in combination modulates benzo(a)pyrene-induced genotoxicity during lung carcinogenesis

The present study explored the effects of combination of Tinospora cordifolia and Arabinogalactan on surface membrane dynamics and programmed cell deaths in rat model of lung cancer. The rats were divided into different groups namely normal control, benzo(a)pyrene (BP) treated, BP + Tinospora cordifolia (TC)-treated, BP + Arabinogalactan (A)-treated and BP + TC + A-treated groups. Significant changes were observed in the membrane dynamics of rats treated with BP. The carcinogen treatment demonstrated a marked decrease in membrane microviscosity. Also, excimer/monomer ratio and fluidity parameters of BP treated rats showed significant rise. On the other hand, combination of Tinospora cordifolia and Arabinogalactan improvised surface membrane dynamics. Moreover, micronuclei formation along with protein expression of bcl-2 showed significant increase in the lungs of BP treated rats. The combined treatment of Tinospora cordifolia and Arabinogalactan moderated the micronuclei formation in BP treated rats. Also, the combined treatment regulated the protein expressions of bcl-2 in BP-treated rats. As a result, marked improvement was noticed in apoptosis of BP treated cells treated with combination treatment. This study concludes that the Tinospora cordifolia and Arabinogalactan in combination improve the surface membrane dynamics and apoptosis in BP-treated rats.

Potential of Ablation Therapy during Hepatocellular Carcinoma

Background: This study explored the potential of ablation therapy on membrane fluidity changes in diethylnitrosamine (DEN) induced hepatic cancer.Methods: Male Wistar rats were segregated into four groups viz., normal control, DEN treated, ablation therapy treated, and DEN ablation therapy treated. We assessed the viscosities as well as fluidity parameters in isolated brush border membranes using the membrane extrinsic fluorophore pyrene.Results: DEN treatment successfully induced hepatic cancer in the livers of rats and ablation therapy worked well in terms of therapy. DEN treatment resulted in a significant rise in lipid peroxidation (LPO) and significant decrease in the, reduced glutathione levels (GSH). A significant decrease was also noticed in the activities of glutathione reductase (GR), glutathione transferase (GST), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) following DEN treatment. On the other hand, ablation therapy treatment to DEN-treated rats resulted in a significant decrease in the LPO levels but caused a significant rise in the levels of GSH. Moreover, the activities of GR, GST, SOD, CAT, and GPx showed significant improvement after ablation therapy treatment. The results further demonstrated a marked decrease in membrane microviscosity following DEN treatment. On the other hand, a significant increase was noticed in both excimer/monomer ratio and fluidity parameter in DEN-treated rats. However, membrane microviscosity and the fluidity alterations were significantly restored back to near normal with ablation therapy treatment.Conclusions: The study, therefore, concluded that ablation therapy holds good therapeutic potential against DEN-induced hepatic cancer.

Azadircta indica as a modulator of membrane stability parameters and surface changes during 1,2 dimethylhydrazine-induced colorectal carcinogenesis

ObjectiveThe aim of the present study was to study the modulatory potential of Azadirta indica on colonic surface abnormalities and membrane fluidity changes following 1,2 dimethylhydrazine-induced [DMH] colon carcinogenesis. Materials and methodsBrush border membranes [BBM] were isolated from the colon of rats and the viscosity as well as fluidity parameters were assessed by using the membrane extrinsic fluorophore pyrene. ResultsDMH treatment resulted in a significant increase in lipid peroxidation [LPO]. Reduced glutathione levels [GSH] and the activities of glutathione reductase [GR], glutathione transferase [GST], superoxide dismutase [SOD], catalase [CAT] and glutathione peroxidase [GPx] were found to be significantly decreased following DMH treatment. On the other hand, supplementation with AI, DMH-treated rats resulted in a significant decrease in the levels of lipid peroxidation but caused a significant increase in the levels of GSH as well in the activities of GR, GST, SOD, CAT and GPx. The results further demonstrated a marked decrease in membrane microviscosity following DMH treatment. On the other hand, a significant increase was observed in the excimer/monomer ratio and fluidity parameter of DMH-treated rats when compared to normal control rats. However, the alterations in membrane microviscosity and the fluidity parameters were significantly restored following A. indica treatment. Further, histological as well as colon surface alterations were also observed following DMH treatment, which however were greatly prevented upon AI co-administration. ConclusionsThe study, therefore, concludes that A. indica proves to be useful in modulating the colonic surface abnormalities and membrane stability following DMH-induced colon carcinogenesis.

Selenium as a modulator of membrane stability parameters and surface changes during the initiation phase of 1,2-dimethylhydrazine induced colorectal carcinogenesis

The present study evaluated the modulatory potential of selenium on colonic surface abnormalities and membrane fluidity changes following 1,2-dimethylhydrazine (DMH) induced colon carcinogenesis. Rats were segregated into four groups viz., normal control, DMH treated, selenium treated, and DMH + selenium treated. Initiation of molecular events leading to colon carcinogenesis was started following weekly subcutaneous injections of DMH (30 mg/Kg body weight) for 10 weeks. Selenium in the form of sodium selenite was supplemented to rats at a dose level of 1 PPM in drinking water, ad libitum for the entire duration of the study. Brush border membranes were isolated from the colon of rats and the viscosity as well as fluidity parameters were assessed using the membrane extrinsic fluorophore pyrene. DMH treatment resulted in a significant increase in lipid peroxidation. Reduced glutathione levels (GSH) and the activities of glutathione reductase (GR), glutathione transferase (GST), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were found to be significantly decreased following DMH treatment. On the other hand, supplementation with selenium to DMH treated rats resulted in a significant decrease in the levels of lipid peroxidation but caused a significant increase in the levels of GSH as well in the activities of GR, GST, SOD, CAT, and GPx. The results further, demonstrated a marked decrease in membrane microviscosity following DMH treatment. On the other hand, a significant increase was observed in the excimer/monomer ratio and fluidity parameter of DMH treated rats when compared to normal control rats. However, the alterations in membrane microviscosity and the fluidity parameters were significantly restored following selenium treatment. Further, histological as well as colon surface alterations were also observed following DMH treatment, which however were greatly prevented upon selenium co-administration. The study, therefore, concludes that selenium proves as a useful in modulating the colonic surface abnormalities and membrane stability following DMH induced colon carcinogenesis.

Membrane Fluidity and Surface Changes During Initiation of 1,2 Dimethylhydrazine-Induced Colon Carcinogenesis: Protection by Zinc

The present study evaluated the modulatory effects of zinc on colonic membrane fluidity and surface abnormalities following 1,2 dimethylhydrazine (DMH)-induced colon carcinogenesis. Rats were segregated into four groups: normal control, DMH treated, zinc treated, DMH + zinc treated. Colon carcinogenesis was initiated through weekly subcutaneous injections of DMH (30 mg/kg body weight) for 8 weeks. Zinc (in the form of zinc sulphate) was supplemented to rats at a dose level of 227 mg/L in drinking water, ad libitum, for the entire duration of the study. Brush border membranes (BBM) were isolated from the colon of rats and the fluidity parameters were assessed by steady-state fluorescence polarization technique using the membrane extrinsic fluorophore 1,6-diphenyl-1,3,5-hexatriene (DPH). The translational diffusion was measured by using the excimer formation of pyrene incorporated in the membrane. The results demonstrated a significant increase in the polarization and anisotropy, accompanied by an increase in order parameter in the membrane preparations from the colon of DMH-injected rats. Further, studies with pyrene fluorophore indicated a marked decrease in membrane microviscosity following DMH treatment. However, the alterations in membrane fluorescence polarization and the fluidity parameters were completely restored following zinc treatment. Drastic alterations in colon surface were noticed after 8 weeks of DMH treatment. However, zinc treatment to DMH-treated rats greatly restored normalcy in the colonic surface. The study concludes that zinc has a strong membrane stabilizing effect and thus has a positive beneficial effect against chemically induced colonic preneoplastic progression in rats.

Sugar coated liposomal flavonoid: A unique formulation in combating carbontetrachloride induced hepatic oxidative damage

Rats were administered a single dose of plant origin phenolic antioxidant Quercetin (QC) in free, liposome encapsulated and galactosylated liposome encapsulated forms 2 h prior to hepatotoxic dose of carbontetrachloride (CCl4, 40% v/v in olive oil, 1 ml/kg b.wt). Among those three different forms of QC tested, only galactosylated liposomal QC provided significant protection against CCl4 induced hepatic oxidative damage. After 24 h of injection (S.C.) hepatic cells of rats were found susceptible to CCl4 induced oxidative damage and it was monitored by the increased amount of conjugated diene in hepatic membrane. The two-fold increase in conjugated diene by the induction of CCl4 was decreased upto normal level by galactosylated liposomal QC pre-treatment.Carbontetrachloride induced membrane damage in hepatic cells and it was judged by the blood serum pathological and liver tissue histopathological examination. Membrane damage by the induction of CCl4 was further evaluated by the decreased level of plasma membrane (PM) bound enzyme Na+/K+ ATPase activity and it was increased only by the pre-treatment of galactosylated liposomal QC.Carbontetrachloride induced a substantial decrease both in enzymatic and molecular endogenous antioxidant levels in hepatic cells.The depression in antioxidant system in hepatic cells was completely prevented by a single dose of galactosylated lipsosomal QC prior to CCl4 treatment.Liver uptake of QC was estimated after 2 h of the flavonoid injection (8.9 μmol/kg body weight) (free or liposomal forms) and 85% of the injected QC was found in liver in the case of galactosylated liposomal QC. Whereas only 25% of the injected dose was detected in liver when an identical amount of free QC was injected.Carbontetrachloride also induced an alteration in membrane fluidity and it was evaluated by a decrease in membrane micro-viscosity. Free QC pre-treatment resulted in no protection against CCl4 induced increase in hepatic membrane fluidity, whereas galactosylated liposomal QC exerted a significant protection against the increase. Results of this study revealed that QC in galactosylated liposome could exert a significant protection against CCl4 induced hepatocellular injury.

Anoxia-Reoxygenation in Cultured Endothelial Cells: Effects of Fructose Diphosphate and Captopril

Cultured rat aortic endothelial cells were morphologically and immunologically characterized before incubation under anoxic conditions for 120 minutes. Cell samples were reoxygenated for 10, 30, and 60 minutes as a model of anoxia-reperfusion injury. The effects of anoxia-reoxygenation were evaluated by measurements of membrane microviscosity, intracellular Ca2+ content, release of 51Cr, and uptake of trypan blue. Membrane microviscosity decreased from 2.03 ± 0.17 poise before anoxia to 1.72 ± 0.22 poise after 120 minutes of anoxia, with a further decrease to 1.54 ± 0.29 poise after 60 minutes of reoxygenation. Release of 51Cr correlated negatively with the decrease in membrane microviscosity and rose from 7.14% ± 0.4% to 12.16% ± 2.79% after anoxia and to 27.17% ± 2.59% after 60 minutes of reoxygenation. Intracellular Ca2+ content and uptake of trypan blue showed no noticeable change during anoxia but they increased significantly during reoxygenation. Addition of fructose-1,6-diphosphate to the anoxic incubation medium partly prevented the change in microviscosity and significantly reduced the release of 51Cr and the uptake of Ca2+ and trypan blue. Captopril exerted similar but less potent effects to those of fructose-1,6-diphosphate.

Anaesthetic potency of inhalation agents is independent of membrane microviscosity.

The decrease in membrane microviscosity of erythrocyte ghosts in the presence of clinically relevant concentrations of seven inhalation anaesthetic agents was studied using fluorescence polarization anisotropy of the membrane incorporated fluorescent probes 1,6-diphenyl-1,3,5-hexatriene and 1-[4-trimethylammoniumphenyl]-6-phenyl-1,3,5-hexatriene. All anaesthetic agents produced a dose-dependent decrease in anisotropy of both probes, indicating decreased membrane microviscosity. The reduction in anisotropy measured at the minimum alveolar concentration (ED50) for anaesthesia was related inversely to the anaesthetic potency of the agent and was directly proportional to the hypothetical concentration of agent in the membrane calculated from lipid-water partition coefficients. These findings do not support the hypothesis that volatile anaesthetic agents act by increasing membrane microviscosity of the bulk lipid bilayer to produce anaesthesia.

A subacute treatment of L-methionine induces an increase in the number of [ 3H]spiperone binding sites in the striatum of the rat

A subacute treatment, 500 mg/kg I.P. twice daily during 5 days, by L-methionine provoked an increase in the B max of [ 3H]-spiperone binding in the striatum of the rat. This increase was associated to a decrease in membrane microviscosity. However in these conditions no changes were found in the [ 3H]-DHA, [ 3H]QNB bindings or in the brain dopamine sensitive adenylate cyclase activity. L-methionine treatment reduced the accumulation of Dopa after NSD 1015 and antagonized the decrease in striatal acetylcholine provoked by haloperidol. Thus L-methionine might be a new potential drug for Parkinson's disease treatment.

Effect of diabetes and insulin replacement on the lipid properties of hepatic smooth endoplasmic reticulum.

This study is a characterization of the lipid properties of the smooth and rough endoplasmic reticulum (SER, RER) of liver from streptozotocin-induced diabetic rats. A significant decrease in membrane microviscosity was observed in the SER but not the RER of diabetic rats when compared to that of normal controls. This decrease in SER membrane microviscosity correlated with a decrease in cholesterol/phospholipid ratio of these membranes that could be accounted for solely by a change in the membrane cholesterol content. Changes in phospholipid fatty acyl chain composition were also observed in the SER membranes but these changes were small when compared to the large change in cholesterol content observed. Insulin treatment for only one day did not significantly alter the microviscosity of the SER but after 2, 4 and 6 days of treatment both membrane microviscosity and membrane cholesterol content were restored to values similar to those for normal animals. No significant changes in the RER lipid composition were observed. It is well known that increases in glucose-6-Pase (G-6-Pase) activity of liver ER membranes are associated with diabetic onset. An increase in the specific activity of G-6-Pase was observed in both SER and RER membrane preparations, although the observed increase in the SER membrane is higher. The changes in the G-6-Pase activity of the SER membranes were correlated with the alterations in the microviscosity and lipid composition of these membranes. It is postulated that lipid properties of the SER membranes may contribute to the regulation of G-6-Pase activity in that membrane.

Changes in phospholipid methyltransferases and membrane microviscosity during induction of rat liver microsomal cytochrome P-450 by phenobarbital and 3-methylcholanthrene.

Rat liver microsomes contained two methyltransferases which converted phosphatidylethanolamine (PE) to phosphatidylcholine (PC). The first methyltransferase converted PE to phosphatidyl-N-methylethanolamine (PME) and the second methyltransferase converted PME to PC. Previous work has shown that increased PME synthesis decreases membrane microviscosity. Therefore, changes in the rat liver microsomal cytochrome P-450, phospholipid methyltransferases and membrane microviscosity after induction by phenobarbital and 3-methylcholanthrene were studied. Phenobarbital and 3-methylcholanthrene increased cytochrome P-450 levels 2- to 3-fold. At low SAM concentration, the proportion of PME among the total phospholipids formed increased significantly, and at a high SAM concentration, the proportion of PC among the total phospholipids formed decreased significantly in microsomes of treated rats. Treatment of rats with phenobarbital and 3-methylcholanthrene also decreased microviscosities of the microsomal membranes and liposomes which were prepared from phospholipids extracted from the microsomes. In synthetic liposomes containing PE, PME and PC, microviscosity decreased when the proportion of PME was increased or the proportion of PC was decreased. These results suggest that the membrane fluidity increases with phenobarbital and 3-methylcholanthrene treatment, and changes in phospholipid methyltransferases may contribute to the process of enzyme induction. During induction with phenobarbital, all three factors known to increase membrane fluidity (linoleic acid content, the formation of phosphatidyl-N-methylethanolamine, and decreases in the cholesterol/phospholipid ratio) contribute to the decrease in microviscosity. During induction with 3-methylcholanthrene, alterations in phospholipid methylation is possibly the primary cause of the decrease in membrane microviscosity.

Dynamic lipid changes in rapidly proliferating hepatic smooth endoplasmic reticulum during acute dexamethasone treatment of adrenalectomized rats.

The regulation by the cell of subcellular membrane components is dependent on a highly complex balance of nutritional, hormonal and metabolic events. We have characterized the lipid components of the endoplasmic reticulum (ER) of the liver of adrenalectomized (ADX) rats and the response of these membrane components to glucocorticoid administration. Membrane microviscosity as measured by fluorescence depolarization of 1,6-diphenylhexatriene (DPH) was measured and correlated with lipid composition and content of the membranes. In the ADX rat, a significant increase in membrane microviscosity of the smooth endoplasmic reticulum (SER) was observed and this was accompanied by an increase in the cholesterol content/mg protein and a decrease in the phospholipid content/mg protein. A change in the fatty acyl chain composition is observed with a significant increase in the mole percentage of arachidonic acid (20:4) and an accompanying decrease in saturated fatty acids. Within 2-6 hr of dexamethasone administration, a decrease in membrane microviscosity is observed that returns this value to one similar to that for normal control animals. Both the cholesterol and the phospholipid contents/mg protein are likewise restored to levels similar to that for control animals beginning at the 2-hr time point. The arachidonic acid and saturated fatty acid content of the constituent phospholipids do not begin to return to values similar to those for control animals until 6 hr after dexamethasone administration. From these experiments, we can conclude that glucocorticoids play a significant regulatory role in determining the lipid properties of rat hepatic microsomal membranes.

Prostaglandin I2 modifies both prolactin binding capacity and fluidity of mouse liver membranes.

The objective of this study was to determine if prostaglandins alter the fluidity of hepatic membranes and if this, in turn, modifies their ability to bind prolactin. Liver homogenates of adult C3H female mice incubated with 1-1000 nM prostaglandin A1, A2, B2, F1 alpha, 6-keto-F1 alpha, E1, E2, I2, or thromboxane B2 provided the 100,000 X g membrane pellets for subsequent ovine prolactin binding and membrane fluidity studies. Only membrane preparations treated with prostaglandin I2 showed an increase in specific binding of ovine prolactin. The effect (40-50%) was maximal at 100 nM prostaglandin I2 after 30 min of incubation and was due to an increase in the number of receptor sites. Under the same conditions, prostaglandin I2 induced a 17% decrease in membrane microviscosity. These data suggest that specific prostaglandins may modulate the number of prolactin receptors in vivo by modifying the fluidity of the lipid bilayer and the subsequent ease with which receptors can assume active configurations within the matrix.

Changes in the membrane microviscosity of mouse red blood cells infected with Plasmodium berghei detected using n-(9-anthroyloxy) fatty acid fluorescent probes.

A set of n-(9-anthroyloxy) fatty acids (n = 2, 6, 9, 12, 16) have been used as fluorescent probes to examine the lipid environment at different depths in the outer membrane of normal mouse erythrocytes and red blood cells from Plasmodium berghei-infected blood. Fluorescent polarization experiments with normal mouse erythrocytes have demonstrated a typical gradient in microviscosity from the surface to the centre of the bilayer as a consequence of the motional properties of the C-atoms of the phospholipid acyl chains. The fluorescent probes rotate faster in the membrane of purified pluriparasitized cells (greater than 90% purity) than with the remaining fraction of red blood cells from infected blood (20--40% immature, infected red cells, and uninfected red cells), or normal mouse erythrocytes. This increase in fluidity with heavily infected cells occurs predominantly at the centre of the lipid bilayer, rather than at the membrane surface. A comparison of the polarization values of intact and lysed infected cells indicates that the fluorescent fatty acids preferentially label the plasma membrane rather than the internal membranes of infected cells. The results suggest that P. berghei infection causes a change in the composition and/or organization of the outer membrane of pluriparasitized cells which produces a decrease in membrane microviscosity.

X-Ray-Induced Structural Changes in Erythrocyte Membranes Studied by Use of Fluorescent Probes

The structural changes in erythrocyte membranes caused by X irradiation were investigated with the aid of fluorescent probes. It was found that the fluorescence intensity of 1-anilino-8-naphthalene sulfonate (ANS) combined with X-irradiated membranes decreased as a function of the dose. The effect could be observed even when 100 R was given. The lowering of the quantum yield of ANS fluorescence in the irradiated membranes seemed to be the primary cause of the fluorescence decrease, as the amount of bound ANS was found to be only slightly decreased. The fluorescence decrease was not accompanied by a significant shift of the emission spectrum to a longer wavelength. In addition, the fluorescence polarization of bound ANS decreased in X-irradiated membranes, suggesting that the decrease in quantum yield of bound ANS might be due to the decrease in membrane microviscosity. Measurement of the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene incorporated into the membranes indicated that it also decre...

Membrane fluidity, capping of cell-surface antigens and immune response in mouse leukaemia cells

Transplantation of primary GRSL cells in the ascitic form led to a decrease in membrane microviscosity as measured by the fluorescence polarization technique. The transplanted GRSL ascitic cells showed a markedly lower ability to form caps with respect to both virus-related (MLr, GIX) and normal (H-2.7(G), H-2.8(K) and TL1.2) cell-surface antigens and their appropriate antisera in the indirect membrane immunofluorescence tests, than did primary GRSL cells, transplanted GRSL cells growing in solid form, and thymocytes, which all exhibited significantly higher membrane microviscosities. Transplantation of primary GRSL cells into syngeneic mice pre-irradiated with 400 rad did not lead to a fall in membrane microviscosity. It is suggested that the host immune response in intact mice leads to a selective survival of ascitic tumour cells with low membrane microviscosity.

Increased membrane fluidity precedes fusion of muscle cells.

EMBRYONIC skeletal muscle cell cultures are highly suitable for demonstrating the dynamic role of the plasma membrane in cellular differentiation. A primary event in the overt development of muscle in culture is fusion of the plasma membranes of neighbouring mononucleated myoblasts to form multinucleated myotubes1,2. Previous studies of muscle fusion have focused on ultrastructural3 and electrophysiological4 aspects, the requirement for Ca2+ (ref. 5), and the importance of fusion for the subsequent biosynthesis of muscle-specific proteins5. But, understanding of this process has been hampered by the absence of reliable markers for the initiation of myoblast fusion4. In addition, the importance of the lipid properties of the cell membrane in regulation of the onset of muscle fusion has not been unambiguously demonstrated in spite of the well-established role of lipids in other cases of membrane fusion6. We examined the possibility that fusion of myoblasts is regulated by changes in the lipid fluidity of the cell membrane, as measured by the rotational diffusion of a fluorescent probe. We report here that, as a primary step in myogenesis in culture, mononucleated myoblasts undergo a sharp decrease in membrane microviscosity, and shortly afterwards fuse rapidly to form multinucleated myotubes. The microviscosity remains at a minimal value during the period of rapid cell fusion. The subsequent post-fusional differentiation of the muscle cell is accompanied by the regeneration of membrane rigidity.