• Archives of Pharmacal Research
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• v.13 no.2
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• pp.192-197
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• 1990

The microviscosites of the lipid bilayers of liposomal membranes of phospholipids were measured by the intermolecular excimer, formation method employing pyrene as a fluorescence probe, and the effects of n-alkanols and other local anesthetics on the microviscosity were investigated. The results showed that the n-alkanols and the ohter local anesthetics effectively lowered the microviscosity of the lipid bilayer of the dipalmitoyl phosphatidycholine liposomal membrane in proportion to the concentration of the additives. Moreover, there was a fairly good correlation between the ocal anesthetic activities and the microviscosity-lowering activities of these drugs. This results suggests that the nerve blocking activity of local anesthetics might have some relation with their activity fluidizing the lipid bilayer of biomembrane.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.3
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• pp.243-251
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• 2000

In order to provide a basis for studying the molecular mechanism of pharmacological action of local anesthetics and to develop a fluorescence spectroscopic method which can detect the microviscosity of native and model membranes using intramolecular excimerization of 1,3-di(l-pyrenyl)propane (Py-3-Py), we examined the effect of lidocaine HCl on the microviscosity of model membranes of phosphatidylcholine fraction extracted from synaptosomal plasma membrane vesicles (SPMVPC). The excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in liquid paraffin was a simple linear function of $T/{\eta}.$ Based on this calibration curve, the microviscosity values of the direct probe environment in SPMVPC model membranes ranged from $234.97{\pm}48.85$ cP at $4^{\circ}C$ to %19.21{\pm}1.11$ cP at $45^{\circ}C.$ At $37^{\circ}C,$ a value of $27.25{\pm}0.44$ cP was obtained. The lidocaine HCl decreased the microviscosity of SPMVPC model membranes in a concentration-dependent manner, with a significant decrease in microviscosity value by injecting the local anesthetic even at the concentration of 0.5 mM. These results indicate that the direct environment of Py-3-Py in the SPMVPC model membranes is significantly fluidized by the lidocaine HCl. Also, the present study explicitly shows that an interaction between local anesthetics and membrane lipids is of importance in the molecular mechanism of pharmacological action of lidocaine HCl.

• Archives of Pharmacal Research
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• v.20 no.1
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• pp.1-6
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• 1997

We determined the microviscosity of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex and liposomes of total lipids (SPMTL) and phospholipids (SPMPL) extracted from SPMV. Changes in the microviscosity induced by the range and rate of lateral diffusion were measured by the intramolecular excimerization of 1, 3-di(1-pyrenyl)propane (Py-3-Py). The microviscosity values of the direct probe environment in SPMV, SPMTL and SPMPL were 38.17, 31.11 and 27.64 cP, respectively, at$37^{\circ}C$and the activation energies $(E_a)$ of the excimer formation of Py-3-Py in SPMV, SPMTL and SPMPL were 8.236, 7.448 amd 7.025 kcal/mol, respectively. Probe location was measured by polarity and polarizability parameters of the probe Py-3-Py and probe analogues, pyrene, 1-pyrenenonanol and 1-pyrenemethyl-3${\beta}$-hydroxy-22, 23-bisnor-5-cholenate (PMC), incorporated into membranes or solubilized in reference solvents. There existed a good linear relationship between the first absorption peak of the $^1_a$ band and the polarizability parameter $(n^{2}-1)/(2n^{2}+1)$.The calculated refractive index values for SPMV, SPMTL and SPMPL were close to 1.50, which is higher than that of liquid paraffin (n=l.475). The probe location was also determined by using a polarity parameter $(f-1/2f^{I})$. Here f=$({\varepsilon}-1)/(2{\varepsilon}+1)$ is the dielectric constant function and $f^I=(n^2-1)/(2n^2+1)$ is the refractive index function. A correlation existed between the monomer fluorescence intensity ratio and the solvent polarity parameter. The probes incorporated in SPMV, SPMTL, and SPMPL report a polarity value close to that of 1-hexanol $({\varepsilon}=13.29)$. In conclusion, Py-3-Py is located completely inside the membrane, not in the very hydrophobic core, but displaced toward the polar head groups of phospholipid molecules, e.g., central methylene region of aliphatic chains of phospholipid molecules.

• The Korean Journal of Pharmacology
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• v.24 no.1
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• pp.43-52
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• 1988

To elucidate the mechanism of action of local anesthetics, the effects of local anethetics on the microenvironment of the lipid bilayers of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine brain and dimyristoylphosphatidylcholine (DMPC) multilamellar liposomes were investigated employing the intermolecular excimer fluorescence technique and differential scanning calorimetry (DSC). The relative intensities of excimer and monomer fluorescence of pyrene are a simple linear function of the viscosity of a homologous series of solvents. The microviscosity(${\eta}$)of the hydrocarbon region of SPMV was measured by this method and the value was $57.3{\pm}5.3\;cP$ at $37^{\circ}C$. In the presence of lidocaine-HCl and procaine-HCl, the values decreased to $46.5{\pm}5.1\;cP$ and $54.7{\pm}4.8\;cP$, respectvely. The differential scanning thermograms of DMPC multilamellar liposomes showed that local anesthetics significantly lowered the phase transition temperature, broadened the thermogram peaks, and reduced the size of the cooperative unit. These results indicate that local anesthetics have significant fluidizing effects on biomembranes and perturbation of membrane lipids may produce some, but not all, of their pharmacological actions.

• Journal of Photoscience
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• v.3 no.1
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• pp.33-38
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• 1996

The effect of vesicular L-$\alpha$-phosphatidylcholine on the rate of formation of all-trans-N-retinylidene-n-butylamine (3) and on the regeneration kinetics of bacteriorhodopsin pigment from retinal and bacterio-opsin have been studied. An estimate of the relative positions of retinal and n-butylamine in the vesicles has been made by fluoresence quenching experiments. Partition coefficient of retinal and microviscosity of the retinal-binding region have also been determined. The results are discussed in terms of the nature of chemical interaction between retinal and amine in a lipid environment.

• Bulletin of the Korean Chemical Society
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• v.17 no.2
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• pp.158-162
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• 1996

The steady-state emission and fluorescence lifetimes of 1,3-dipyrenylpropane were measured in silicate sol-gel and xerogel matrices. In sol solution, the fluorescence emission spectra of monomer and excimer resemble those in hydrocarbon solvents. In gel and xerogel condition, however, the fluorescence spectra exhibit significant change, largely confirming the intramolecular motions in gel pores are influenced by microviscosity. The rate constants for intramolecular excimer formation were obtained from the measured fluorescence lifetimes and the rate processes for excimer forming in silicate sol-gel are described by a simple kinetic scheme.

• Macromolecular Research
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• v.15 no.4
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• pp.385-391
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• 2007

Block copolymer micelles are generally formed via the self-assembly of amphiphilic block copolymers in an aqueous medium. The hydrophilic and hydrophobic blocks form shell and core micelles, respectively. The block copolymers of methoxy poly(ethylene glycol) (MPEG)-b-poly(2-diisopropylamino)ethyl methacrylate (PDPA) were synthesized via atom transfer radical polymerization, with the macro initiator synthesized by the coupling of 2-bromoisobutyryl bromide with MPEG in the presence of a triethyl amine base catalyst. The atom transfer radical polymerization of 2-diisopropylamino)ethyl methacrylate was performed in conjunction with an N,N,N',N",N"-pentamethyl-diethylenetriamine/copper bromide catalyst system, in DMF, at $70^{\circ}C$. The pH induced micellization/demicellization was studied using fluorescence, with a pyrene probe. Furthermore, the pH dependent micellization was confirmed using the microviscosity method, with a dipyme fluorescence probe. The pH dependant micelle size distribution was studied using dynamic light scattering. The characterization of the synthesized polymers was established using gel permeation chromatography and from the $^1H-nuclear$ magnetic resonance spectroscopy.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.8
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• pp.1220-1225
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• 2008

This study was conducted to investigate the protective effect of genistein on the antioxidative defence system and membrane fluidity in chick skeletal muscle cells after supplementation with 0, 20, 40, and $80{\mu}mol/L$ genistein in $50{\mu}mol/L$ $FeSO_4/H_2O_2$ treated cells for 24 h. Genistein supplementation recovered the decreased activity of total superoxide dismutase induced by $FeSO_4/H_2O_2$, significantly increased glutathione peroxidase activity (p<0.05) and decreased malondialdehyde production (p<0.05). The treatment of 80 mol/L genistein in $FeSO_4/H_2O_2$ treated cells decreased the secretion of creatine kinase (p<0.05). Fluorescence polarization values and microviscosities observed with $FeSO_4/H_2O_2$ treated cells were significantly higher than those observed with no $FeSO_4/H_2O_2$ treated cells. The addition of $80{\mu}mol/L$ genistein improved the increased fluorescence polarization value (p<0.05) caused by $FeSO_4/H_2O_2$ treatment. The microviscosity value was significantly decreased by adding genistein (p<0.05). In conclusion, genistein protected skeletal muscle cells from oxidative damage by improving antioxidative status and membrane fluidity.

• Applied Chemistry for Engineering
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• v.9 no.3
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• pp.457-461
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• 1998

The vesicle system of DPPC(dipalmitoylphosphaticylcholine)/Chol(Cholesterol) has been modified by incorporating various mole fractions of flourinated surfactant($C_8F_{17}(CH_2)_2OCO-CH_2CH(SO_3Na)COO(CH_2)_2C_8F_{17}$. Sodium bis(1H,1H,2H,2H-heptadecaflurododecyl)-2-sulfosuccinate, FS)/fluorinated fatty acid salt ($C_7F_{15}COONH_4$, ammoniumpentadecaflurooctyrate, FFS), and their physicochemical properties have been investigated in an attempt to enhance the stability of phospholipid vesicle system. The ${\zeta}$-potential measurement by use of Zetamaster sub-micron Particle Electrophoresis Analyzer (Malvern Co.) showed that a charged homogeneous DPPC/Chol/FS vesicle has been formed owing to the incorporated FFS effect on the membrane, playing a role as a cosurfactant in the bilayer between DPPC and FS components. With increase in the concentration of FFS, it was found that the particle size and also surface charge of the DPPC/Chol/FS vesicle decreased. The stability of DPPC/Chol/FS/FFS liposome was found to be enhanced significantly compared to that of DPPC/Chol/FS according to the dispersity change as a function of time. The release rate of dye molecule of Methylene Blue from the DPPC/Chol/FS/FFS vesicle was determined to be slower than that of DPPC/Chol/FS system, and it may be attributed to the increase in microviscosity of the hydrophobic region in the bilayer. The affinfinity of DPPC/Chol/FS/FFS vesicles to albumin was found to be slightly lowered compared to that of DPPC/Chol/FS. Based on these findings, it was confirmed that a more stable and homogeneous vesicle system of DPPC/Chol/FS could be prepared by addition of FFS, acting as a cosurfactant in the aggregate formation.