Cannabidiol and Cannabigerol Modify the Composition and Physicochemical Properties of Keratinocyte Membranes Exposed to UVA.

The action of UVA radiation (both that derived from solar radiation and that used in the treatment of skin diseases) modifies the function and composition of keratinocyte membranes. Therefore, this study aimed to assess the effects of phytocannabinoids (CBD and CBG), used singly and in combination, on the contents of phospholipids, ceramides, lipid rafts and sialic acid in keratinocyte membranes exposed to UVA radiation, together with their structure and functionality. The phytocannabinoids, especially in combination (CBD+CBG), partially prevented increased levels of phosphatidylinositols and sialic acid from occurring and sphingomyelinase activity after the UVA exposure of keratinocytes. This was accompanied by a reduction in the formation of lipid rafts and malondialdehyde, which correlated with the parameters responsible for the integrity and functionality of the keratinocyte membrane (membrane fluidity and permeability and the activity of transmembrane transporters), compared to UVA-irradiated cells. This suggests that the simultaneous use of two phytocannabinoids may have a protective effect on healthy cells, without significantly reducing the therapeutic effect of UV radiation used to treat skin diseases such as psoriasis.

Antimicrobial Activity of Quercetin, Naringenin and Catechin: Flavonoids Inhibit Staphylococcus aureus-Induced Hemolysis and Modify Membranes of Bacteria and Erythrocytes.

Search for novel antimicrobial agents, including plant-derived flavonoids, and evaluation of the mechanisms of their antibacterial activities are pivotal objectives. The goal of this study was to compare the antihemolytic activity of flavonoids, quercetin, naringenin and catechin against sheep erythrocyte lysis induced by α-hemolysin (αHL) produced by the Staphylococcus aureus strain NCTC 5655. We also sought to investigate the membrane-modifying action of the flavonoids. Lipophilic quercetin, but not naringenin or catechin, effectively inhibited the hemolytic activity of αHL at concentrations (IC50 = 65 ± 5 µM) below minimal inhibitory concentration values for S. aureus growth. Quercetin increased the registered bacterial cell diameter, enhanced the fluidity of the inner and surface regions of bacterial cell membranes and raised the rigidity of the hydrophobic region and the fluidity of the surface region of erythrocyte membranes. Our findings provide evidence that the antibacterial activities of the flavonoids resulted from a disorder in the structural organization of bacterial cell membranes, and the antihemolytic effect of quercetin was related to the effect of the flavonoid on the organization of the erythrocyte membrane, which, in turn, increases the resistance of the target cells (erythrocytes) to αHL and inhibits αHL-induced osmotic hemolysis due to prevention of toxin incorporation into the target membrane. We confirmed that cell membrane disorder could be one of the direct modes of antibacterial action of the flavonoids.

Changes in the Physicochemical Properties of Blood and Skin Cell Membranes as a Result of Psoriasis Vulgaris and Psoriatic Arthritis Development.

Psoriasis is accompanied by disturbed redox homeostasis, with systemic and local oxidative stress promoting the modification of basic components of cellular membranes. Therefore, the aim of the study was to investigate the effect of development of psoriasis vulgaris and psoriatic arthritis on the composition and physicochemical properties of skin cell membranes (keratinocytes and fibroblasts) and blood cells (lymphocytes, granulocytes and erythrocytes). Both forms of psoriasis are characterized by decreased levels and changes in the localization of membrane phospholipids, and an increased level of sialic acid as well as the lipid peroxidation product (malondialdehyde), which resulted in an increase in the zeta potential of skin cells and blood cells, with granulocytes and lymphocytes affected more than erythrocytes. Using theoretical equations and the dependence of the cell membrane surface charge density as a function of pH, it was shown that patients with psoriatic arthritis have a greater increase in the concentration of negatively charged groups on the membrane surface and reduced the value of the association constant with H+ compared to patients with psoriasis vulgaris. Therefore, it can be suggested that the physicochemical parameters of membranes, skin and blood cells, especially lymphocytes, can be used to assess the severity of the disease.

Cannabidiol-Mediated Changes to the Phospholipid Profile of UVB-Irradiated Keratinocytes from Psoriatic Patients.

UVB phototherapy is treatment for psoriasis, which increases phospholipid oxidative modifications in the cell membrane of the skin. Therefore, we carried out lipidomic analysis on the keratinocytes of healthy individuals and patients with psoriasis irradiated with UVB and treated with cannabidiol (CBD), phytocannabinoid with antioxidant and anti-inflammatory properties. Our results showed that, in psoriatic keratinocytes phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidylserine (PS), and ether-linked phosphoethanolamine (PEo), were downregulated, while SM (d41:2) was upregulated. These changes were accompanied by an increase in negative zeta potential, which indicates translocation of PS to the outer layer of the membrane. CBD treatment of psoriatic keratinocytes led to downregulation of PC, PS, and upregulation of certain PEo and an SM species, SM (d42:2), and the zeta potential. However, UVB irradiation of psoriatic keratinocytes resulted in upregulation of PC, PC plasmalogens (PCp), PEo, and a decrease in the negative zeta potential. The exposure of UVB-irradiated cells to CBD led to a decrease in the level of SM (d42:2). Our results suggest that CBD induces pro-apoptotic mechanisms in psoriatic keratinocytes while simultaneously improving the antioxidant properties and preventing the loss of transepidermal water of keratinocytes of patients irradiated with UVB. Thus, CBD has potential therapeutic value in the treatment of psoriasis.

Effects of hypertension and FAAH inhibitor treatment of rats with primary and secondary hypertension considering the physicochemical properties of erythrocytes.

Hypertension is one of the most common cardiovascular diseases in the world and is associated with oxidative stress. The aim of this study was to examine the effect of the chronic administration of the fatty-acid amide hydrolase inhibitor (URB597-[3-(3-carbamoylphenyl)phenyl]N-cyclohexylcarbamate) to rats with primary (SHRs - spontaneously hypertensive rats) and secondary (DOCA-salt - 11-desoxycorticosterone acetate-salt-induced hypertension) hypertension on the composition and physicochemical properties of erythrocytes membrane. Because changes in membrane composition lead to modifications of electrical charge what may affect cell functions, the levels of following components were determined: four classes of membrane phospholipids (by HPLC - high-performance liquid chromatograph), sialic acid (by resorcinol method), lipid peroxidation product - malondialdehyde (by GCMS - gas chromatography-mass spectrometry). The reduced levels of phospholipids and sialic acid, as well as the increased levels of malonodialdehyde observed in the erythrocyte membrane of rats with primary and secondary hypertension led to a decrease in the negative electrical charge of the membrane. Long-term administration of URB597 to SHRs and DOCA-salt-treated rats partially prevented changes caused by hypertension. Using theoretical equations and the dependence of cell surface charge density as a function of pH, total surface concentrations of acid and base groups and their association constants have been determined. Considering the changes in physicochemical parameters of erythrocyte membranes, URB597 can be considered a potential protective factor for erythrocytes in situations of metabolic changes associated with oxidative stress.

Association equilibria of divalent ions on the surface of liposomes formed from phosphatidylcholine.

Divalent ions, in particular calcium ions, constitute important macroelements in living organisms. They are also found in cell membranes, i.e., ensuring their stabilization or participating in synaptic transmission of nerve impulses. The aim of this work is to describe the interactions of divalent ions, such as Ca2+, Ba2+, and Sr2+, in electrolytes with the functional groups on the surface of liposomes formed from phosphatidylcholine (PC). Microelectrophoresis is used to determine the surface charge density as a function of pH. The interactions between ions found in solution and the functional groups of PC are described with the use of a seven-equilibrium mathematical model. Using this model along with experimental data on the charge density of the membrane surface, the association constants characterizing this equilibrium are determined. These parameters are used to calculate the theoretical model curves. The validity of the proposed model is confirmed by comparing the theoretically calculated changes in charge density on the liposome surface with the experimental results.

Association of alkali metal cations with phosphatidylcholine liposomal membrane surface.

Interactions of alkali metal cations (Li+, Na+, K+, Cs+) with phosphatidylcholine (PC) liposomal membranes were investigated through experimental studies and theoretical considerations. Using a microelectrophoresis technique, charge densities of experimental membrane surfaces were measured as a function of the pH of electrolyte solutions. Equilibria between the PC liposomal membranes and monovalent ions were mathematically analyzed and described quantitatively through a previously proposed theoretical model. Association constants between functional groups of PC and the studied ions were determined and used to define theoretical curves of membrane surface charge density versus pH. Theoretical and experimental data were compared to verify the model. The PC membrane was found to have the highest affinity for lithium ions, among the ions tested.

Effects of UVB Radiation on the Physicochemical Properties of Fibroblasts and Keratinocytes.

The skin is the largest human organ, providing the first line of defense to protect the body from physical and environmental effects. The aim of this study was to determine the influence of short-wave ultraviolet (UVB) radiation on the membrane electrical properties, phospholipid content, and lipid peroxidation levels of fibroblasts and keratinocytes. Changes in cell function may affect the basal electrical surface properties of cell membranes. These changes can be detected using electrokinetic measurements. In this study, the surface charge densities of fibroblasts and keratinocytes were measured as a function of pH. A four-component equilibrium model was used to describe the interaction between the ions in solution and on cell membrane surfaces. Agreement was found between the experimental and theoretical charge variation curves of leukemia cells from pH 2.5 to pH 9. Phospholipid composition was determined qualitatively and quantitatively by HPLC, and lipid peroxidation was estimated by measuring the level of malondialdehyde. The acidic functional group concentrations and average association constants with hydroxyl ions were higher, and the average association constants with hydrogen ions were smaller in UVB-treated skin cell membranes compared to those in untreated cells. Moreover, our results showed that UVB radiation is associated with increased levels of phospholipids and lipid peroxidation products in fibroblasts and keratinocytes.

Characterization of human bladder cell membrane during cancer transformation.

Phenomena associated with changes in cell membranes are thought to play an important role in the cancer transformation. We hypothesized that the electrical charge of tumor cells can indirectly represent membrane-based changes that have occurred during cell transformation and may indicate tumor cell status. Here, we describe work showing that phospholipids, proteins content, and electric charge, are all altered in the cell membranes of pT2 stage/grade G3 bladder cancer. Qualitative and quantitative phospholipid composition and the presence of integral membrane proteins were identified using high-performance liquid chromatography. Protein composition was determined using selective hydrolysis of isolated bladder cell membrane proteins and peptide resolution. The surface charge density of human bladder cell membranes was determined using electrophoresis. Our results show that cancer transformation is associated with increased phospholipid levels and a decreased level of integral proteins. Moreover, the process of cancer transformation significantly enhanced changes in the surface charge density of the human bladder cell membrane. In conclusion, this study demonstrates that cell membrane structure and function are modified in bladder cancer cells and that further work in this area is warranted.

Effects of novel dinuclear cisplatinum(II) complexes on the electric properties of human breast cancer cells.

The aim of this study was to determine the influence of cisplatin and novel dinuclear platinum(II) complexes on the electrical properties of the membrane and the level of lipid peroxidation in the human breast cancer cell lines MDA-MB-231 and MCF-7. The basal electrical surface properties of cells are known. Changes in cell function may affect these surface properties, and those changes can be detected by electrokinetic measurements. The surface charge density of the breast cancer cell lines MDA-MB-231 and MCF-7 were measured as a function of pH. A four-component equilibrium model was used to describe the interaction between the solution ions and the breast cancer cell surface. The experimental and the theoretical charge variation curves of the breast cancer cells at pH 2.5-9 were in agreement. Measurements of the cellular malondialdehyde levels with high performance liquid chromatography were used to determine the extent of lipid peroxidation. The acid and base functional group concentrations and average association constants with hydroxyl ions were smaller in breast cancer cell membranes treated with cisplatin or novel dinuclear platinum(II) complexes compared with untreated cancer cells, and the average association constants with hydrogen ions were higher. The levels of lipid peroxidation products in breast cancer cells treated with cisplatin or novel dinuclear platinum(II) complexes were also higher than in untreated cancer cells.

Changes in electric properties of human breast cancer cells.

Studies of the electrical surface properties of biological cells have provided fundamental knowledge about the cell surface. The change in biological functions of cells may affect the surface properties and can be detected by electrokinetic measurements. The surface density of fibroblasts and breast cancer cells (MDA-MB-231 and MCF-7) as a function of pH was measured by electrophoresis. The interaction between solution ions and the breast cancer cell or fibroblast surface was described by a four-component equilibrium model. The agreement between the experimental and theoretical charge variation curves of the breast cancer cells and fibroblasts was good at pH 2.5-9. The extent of fibroblast and breast cancer cell lipid peroxidation was estimated by HPLC measurement of the malondialdehyde level. The acid (C(TA)) and basic (C(TB)) functional group concentrations and the average association constant with hydroxyl (K(BOH)) ions values of the breast cancer cell membranes were higher than in normal cells, while the average association constant with hydrogen (K(AH)) value was smaller. The level of lipid peroxidation products was higher in breast cancer cells than in normal cells.

Phospholipid composition and electric charge in healthy and cancerous parts of human kidneys.

Phospholipids are ubiquitous in nature and are essential for the lipid bilayer of cell membranes. Their structural and functional properties are pivotal for the survival of the cell. In this study the phospholipids of healthy and cancerous human renal tissues from the same patients are compared with special reference to the electric charge of the membrane. A simple and highly effective normal-phase method is described for analyzing phospholipids content. This work is focused on changes of phospholipids content (PtdIns, phosphatidylinositol; PtdSer, phosphatidylserine; PtdEtn, phosphatidylethanoloamine; PtdCho, phosphatidylcholine) in cell membranes of renal cancer of pT1 stage, G2 grade, without metastasis. Surface charge density of healthy and cancerous human renal tissues was measured by electrophoresis. The measurements were carried out at various pH of solution. Depending on the surface charge density as a function of pH, acidic (C(TA)) and basic (C(TB)) functional group concentrations and their average association constants with hydrogen (K(AH)) or hydroxyl (K(BOH)) ions were evaluated. The process of cancer transformation was accompanied by an increase in total amount of phospholipids as well as an increase in C(TA) and K(BOH), whereas K(AH) and C(TB) were decreased compared with unchanged tumor cells.

Differences in the phospholipid profile of melanocytes and melanoma cells irradiated with UVA and treated with cannabigerol and cannabidiol.

UV radiation inducing mutations in melanocytes might cause melanoma. As changes in lipid composition and metabolism are associated with many types of cancer including skin cancer, we aimed to evaluate the effects of two phytocannabinoids cannabidiol (CBD) and cannabigerol (CBG), on changes in phospholipid and ceramide (CER) profiles induced by UVA irradiation in human melanocytes and melanoma. UVA radiation caused a significant up-regulation PC, PI and SM species and decrease of CERs content in both types of cells, while up-regulation of PEo was only observed in melanocytes. Exposure of UVA-irradiated melanocytes or melanoma cells to CBD and/or CBG led to significant decrease in relative content of PC, PI and SM specie; however, this effect was more pronounced in cancer cells. Interestingly, only in UVA-irradiated melanocytes and not in melanoma, PEo content was lowered after CBD treatment, while CBG led to additional up-regulation of PEo species. CBD and CBG used together caused decrease of zeta potential, inhibiting PS externalization, and different changes in relative contents of CER and SM species of irradiated and non-irradiated melanoma cells. Obtained results are quite promising due to CBD and CBG abilities to partial reverse pro-cancerogenic changes in phospholipid and CER profiles induced by UVA.

Protective effect of blackcurrant on liver cell membrane of rats intoxicated with ethanol.

Chronic ethanol intoxication oxidative stress participates in the development of many diseases. Nutrition and the interaction of food nutrients with ethanol metabolism may modulate alcohol toxicity. One such compound is blackcurrant, which also has antioxidant abilities. We investigated the effect of blackcurrant as an antioxidant on the composition and electrical charge of liver cell membranes in ethanol-intoxicated rats. Qualitative and quantitative phospholipid composition and the presence of integral membrane proteins were determined by high-performance liquid chromatography. Electrophoresis was used to determine the surface charge density of the rat liver cell membranes. Ethanol intoxication is characterized by changes in cell metabolism that alter the structure and function of cell membrane components. Ethanol increased phospholipid levels and altered the level of integral proteins as determined by decreased phenylalanine, cysteine, and lysine. Ethanol significantly enhanced changes in the surface charge density of the liver cell membranes. Administration of blackcurrant to rats intoxicated with ethanol significantly protected lipids and proteins against oxidative modifications. It is possible that the beneficial effect of blackcurrant is connected with its abilities to scavenge free radicals and to chelate metal ions.

Hydrolysable tannins change physicochemical parameters of lipid nano-vesicles and reduce DPPH radical - Experimental studies and quantum chemical analysis.

Tannins belong to plant secondary metabolites exhibiting a wide range of biological activity. One of the important aspects of the realization of the biological effects of tannins is the interaction with lipids of cell membranes. In this work we studied the interaction of two hydrolysable tannins: 1,2,3,4,6-penta-O-galloyl-ß-d-glucose (PGG) and 1,2-di-O-galloyl-4,6-valoneoyl-ß-d-glucose (T1) which had the same number of both aromatic rings (5) and hydroxyl groups (15) but differing in flexibility due to the presence of valoneoyl group in the T1 molecule with DMPC (dimyristoylphosphatidylcholine) lipid nano-vesicles (liposomes). Tannins-liposomes interactions were investigated using fluorescence spectroscopy, differential scanning calorimetry, laser Doppler velocimetry, dynamic light scattering and Fourier Transform Infra-Red spectroscopy. It was shown that more flexible PGG molecules stronger decreased the microviscosity of the liposomal membranes and increased the values of negative zeta potential in comparison with the more rigid T1. Both compounds diminished the phase transition temperature of DMPC membranes, interacted with liposomes via PO groups of head of phospholipids and their hydrophobic regions. These tannins neutralized DPPH free radicals with the stoichiometry of the reaction equal 1:1. The effects of the studied compounds on liposomes were discussed in relation to tannin quantum chemical parameters calculated by molecular modeling.

The Differential Effect of Cannabidiol on the Composition and Physicochemical Properties of Keratinocyte and Fibroblast Membranes from Psoriatic Patients and Healthy People.

The development of psoriasis is accompanied by oxidative stress, which can modify the components of skin cells. Therefore, the aim of this study was to evaluate the effect of cannabidiol (CBD), an antioxidant and anti-inflammatory phytocannabinoid, on the composition and physicochemical properties of the membranes of healthy and psoriatic keratinocytes and fibroblasts exposed to ultraviolet A (UVA) and ultraviolet B (UVB) radiation. In psoriasis-altered cells, decreased levels of the main groups of phospholipids and increased levels of sialic acid and malondialdehyde (MDA), a lipid peroxidation product, as well as negative charge of cell membranes compared to non-diseased cells, were found. On the other hand, UVA/B radiation increased the levels of phospholipids and MDA in both groups of cells. Moreover, psoriatic cells were characterized by lower levels of sialic acid and negative charge of cell membranes, while non-diseased cells showed the opposite response. The CBD treatment intensified some of the changes (phospholipid content and membrane charge) caused by the radiation of psoriatic cells, while it prevented these changes in the cells of healthy people. The results of this study indicate that CBD can prevent structural and functional changes to the membranes of healthy skin cells during phototherapy for psoriasis.

Inhibition of interaction between Staphylococcus aureus α-hemolysin and erythrocytes membrane by hydrolysable tannins: structure-related activity study.

The objective of the study was a comparative analysis of the antihemolytic activity against two Staphylococcus aureus strains (8325-4 and NCTC 5655) as well as α-hemolysin and of the membrane modifying action of four hydrolysable tannins with different molecular mass and flexibility: 3,6-bis-O-di-O-galloyl-1,2,4-tri-O-galloyl-ß-D-glucose (T1), 1,2,3,4,5-penta-O-galloyl-ß-D-glucose (T2), 3-O-galloyl-1,2-valoneoyl-ß-D-glucose (T3) and 1,2-di-O-galloyl-4,6-valoneoyl-ß-D-glucose (T4). We showed that all the compounds studied manifested antihemolytic effects in the range of 5-50 µM concentrations. However, the degree of the reduction of hemolysis by the investigated tannins was not uniform. A valoneoyl group-containing compounds (T3 and T4) were less active. Inhibition of the hemolysis induced by α-hemolysin was also noticed on preincubated with the tannins and subsequently washed erythrocytes. In this case the efficiency again depended on the tannin structure and could be represented by the following order: T1 > T2 > T4 > T3. We also found a relationship between the degree of antihemolytic activity of the tannins studied and their capacity to increase the ordering parameter of the erythrocyte membrane outer layer and to change zeta potential. Overall, our study showed a potential of the T1 and T2 tannins as anti-virulence agents. The results of this study using tannins with different combinations of molecular mass and flexibility shed additional light on the role of tannin structure in activity manifestation.

Changes in physicochemical properties of kidney cells membrane as a consequence of hypertension and treatment of hypertensive rats with FAAH inhibitor.

Hypertension is a civilization disease leading to remodeling and damage of blood vessels, impaired renal function and premature death. The aim of this study was to compare the effect of chronic administration of URB597, the FAAH (fatty acid amide hydrolase) inhibitor, to rats with primary (SHRs) and secondary (DOCA-salt hypertensive rats) hypertension on electrical and physicochemical properties of kidney cells membranes. Changes in the electrical charge of the membrane may affect the cell functions. The electrical properties of the kidney cells (surface charge density, zeta potential) were measured by electrophoresis. Qualitative and quantitative composition of the membrane (phospholipids and proteins) was determined by HPLC and lipid peroxidation product (4-hydroxy-2E-hexenal; 4-HHE) level was examined by GCMSMS, while the sialic acid content was measured by resorcinol method. In rats with primary hypertension (SHR) and secondary hypertension (DOCA-salt), changes in electrical properties (increase of electric charge and zeta potential) and membrane composition (increase in sialic acid and protein concentration and decrease in phospholipid level) of kidney cells are observed in comparison to control animals. Greater changes were observed in DOCA-salt hypertensive rats. Changes in membrane properties caused by URB597 depend on the type of hypertension. The administration of URB597 to rats with primary hypertension partially prevents changes in the electrical properties (electrical charge, zeta potential) of the membrane caused by hypertension as well as in the sialic acid and proteins content. However, there is no reduction in oxidative stress, assessed by the level of 4-HHE, which may affect the metabolic function of the kidneys. URB597 administered to rats with DOCA salt does not prevent, but rather intensifies, changes caused by hypertension in the kidney. In conclusion, URB597 given to individuals with hypertension, particularly with secondary hypertension, enhancing some disturbances in electric and physicochemical properties of kidney cells observed in hypertension what may lead to additional kidney disorders. Therefore, further researches are necessary.

Effect of monovalent ion adsorption on the electric charge of phosphatidylcholine - decylamine liposomal membranes.

We examined the effect of adsorbed monovalent ions on the surface charge of phosphatidylcholine (PC) - decylamine (DA) liposomal membranes. Surface charge density values were determined from electrophoretic mobility measurements of lipid vesicles performed at various pH levels. The interaction between solution ions and the PC-DA liposomal surface was described by a six component equilibrium model. The previously determined association constants of the -PO((-)) and -N((+))(CH(3))(3) groups of PC with H(+), OH(-), Na(+) and Cl(-) ions (K (A1H), K (B1OH), K (A1Na), K (B1C1)) were used to calculate K (B2OH), and K (B2C1), the association constants of the -N((+))H(3) group of DA with OH(-) and Cl(-) ions, providing an experimental verification for the proposed model.

Changes in Phospholipid Composition Studied by HPLC and Electric Properties of Liver Cell Membrane of Ethanol-Poisoned Rats.

Ethanol introduced into the organism undergoes rapid metabolism to acetaldehyde and then to acetic acid. The process is accompanied by formation of reactive oxygen species (ROS), which damage mainly lipids of membrane cells. The effects of ROS can be neutralized by administering preparations with antioxidant properties. The natural preparations of this kind are teas.This paper reports data on the effect of green and black tea on the surface charge density, content of phospholipids, and level of lipid peroxidation products of liver cell membrane of rats chronically intoxicated with ethanol. Surface charge density of liver cells was measured by the electrophoresis method, whereas qualitative phospholipid composition was determined by the HPLC method.Ethanol administration caused an increase in the amount of all phospholipids, in surface charge density as well as in lipid peroxidation products. Ingestion of green and black tea with ethanol partially prevented these ethanol-induced changes, and the action of green tea was stronger than that of black tea.