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.

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.

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.

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.

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.

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.

Effect of sweet grass (Hierochloe odorata) on the physico-chemical properties of liver cell membranes from rats intoxicated with ethanol.

Changes in the composition and physicochemical properties of liver cell membranes due to ethanol intoxication are due mainly to reactive oxygen species (ROS). The destructive action of free radicals can be neutralized by administration of antioxidants. The purpose of this study was to investigate the efficacy of sweet grass on the physicochemical and biochemical properties of the rat liver membrane altered by chronic ethanol intoxication. Qualitative and quantitative composition of phospholipids and proteins in the membrane were determined by HPLC. 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 as determined by electrophoresis. Administration of sweet grass to rats intoxicated with ethanol significantly protects lipids and proteins against oxidative modifications. Therefore, sweet grass protects against some of the deleterious membrane changes associated with ethanol exposure.

Protective effect of black tea on integral membrane proteins in rat liver.

Ethanol intoxication is accompanied by oxidative stress formation. Consequently, it leads to disturbances in cellular metabolism that can alter the structure and function of cell membrane components. Black tea displays antioxidant properties, protects membrane phospholipids and may protect integral membrane proteins. In the present study, we examined whether black tea induces changes in the liver integral membrane proteins of 12-months old rats chronically intoxicated with ethanol. To estimate qualitatively and quantitatively the levels of the liver integral membrane proteins, the proteins were selectively hydrolyzed by trypsin, the obtained peptides were resolved by HPLC and the levels of specific amino acids within the individual peptides were determined. All of the obtained peptides contained phenylalanine (Phe), cysteine (Cys) and lysine (Lys). Compared to the control group, rats in the ethanol intoxication group showed decreased liver levels of integral membrane proteins as well as fewer trypsin-hydrolyzed peptides and amino acids in the hydrolyzed peptides. Administration of black tea to ethanol-intoxicated rats partially protected proteins against the structural changes caused by ethanol. Black tea prevented decreases in the levels of cysteine (in about 90% of cases), lysine (in about 60% of cases), phenylalanine (in about 70% of cases) and examined peptides (in about 60% of cases). The liver protein level was higher (by about 18%) in rats who received black tea and ethanol than in those who received ethanol alone. In conclusion, black tea partially protects the composition and level of rat liver cell integral membrane proteins against changes caused by ethanol intoxication.

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.

Effect of L-carnitine on liver cell membranes in ethanol-intoxicated rats.

Ethanol intoxication is characterized by changes in cell metabolism which alter the structure and function of cell membrane components, including phospholipids and integral membrane proteins. The interaction of food nutrients with ethanol may modulate alcohol toxicity. One such compound is l-carnitine (l-3-hydroxy-4-N,N,N-trimethylaminobutyrate), which is also an antioxidant. Here we investigate l-carnitine as an antioxidant and assess its effect 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 (HPLC). Electrophoresis was used to determine the surface charge density of the rat liver cell membranes. Ethanol increased phospholipid levels and altered the level of integral proteins as determined by decreased phenylalanine (Phe), cysteine (Cys) and lysine (Lys). Ethanol significantly enhanced changes in the surface charge density of the liver cell membranes. l-Carnitine administration to ethanol-intoxicated rats significantly protects phospholipids and proteins against oxidative modifications. Therefore, the beneficial effect of l-carnitine may be connected to its ability to scavenge free radicals.

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.

Altered membrane free unsaturated fatty acid composition in human colorectal cancer tissue.

Polyunsaturated free fatty acids (PUFAs) participate in normal functioning of the cell, particularly in control intracellular cell signalling. As nutritional components they compose a human diet with an indirect promoting influence on tumourogenesis. The PUFAs level depends on the functional state of the membrane. This work is focused on changes only of free unsaturated fatty acids amount (AA - arachidonic acid, LA - linoleic acid, ALA - alpha-linolenic acid, palmitoleic acid (PA) and oleic acid) in cell membranes of colorectal cancer of pT3 stage, G2 grade without metastasis. Qualitative and quantitative composition of free unsaturated fatty acids in the membrane was determined by high-performance liquid chromatography. It was shown that the malignant transformation was accompanied by a decrease in amount of LA and ALA while arachidonic and oleic acids increased. It is of interest that free AA levels are elevated in colon cancer, as AA is the precursor to biologically active eicosanoids.

Changes in electric charge and phospholipids composition in human colorectal cancer cells.

Cancer cells perform their malicious activities through own cell membranes that screen and transmit inhibitory and stimulatory signals out of the cells and into them. This work is focused on changes of phospholipids content (PI-phosphatidylinositol, PS-phosphatidylserine, PE-phosphatidylethanolamine, PC-phosphatidylcholine) and electric charge that occur in cell membranes of colorectal cancer of pT 3 stage, various grades (G2, G3) and without/with metastasis. Qualitative and quantitative composition of phospholipids in the membrane was determined by HPLC (high-performance liquid chromatography). The surface charge density of colorectal cancer cell membranes was measured using electrophoresis. The measurements were carried out at various pH of solution. It was shown that the process of cancer transformation was accompanied by an increase in total amount of phospholipids as well as an increase in total positive charge at low pH and total negative charge at high pH. A malignant neoplasm cells with metastases are characterized by a higher PC/PE ratio than malignant neoplasm cells without metastases.

Protective effect of green tea on erythrocyte membrane of different age rats intoxicated with ethanol.

It is known that aging is characterized by changes in cell metabolism resulting in modification of the structure and function of cell membrane components which is mainly the consequence of reactive oxygen species action. These disturbances are also enhanced by different xenobiotics, e.g. ethanol. Therefore, the aim of this paper is to examine green tea influence on total antioxidant status (TAS) and on composition and electric charge of erythrocyte membrane phospholipids in ethanol intoxicated rats of various ages. Antioxidant abilities of erythrocytes were estimated by measuring TAS. Qualitative and quantitative composition of phospholipids in the membrane was determined by HPLC, while the extent of erythrocytes lipid peroxidation was estimated by HPLC measurement of malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) levels. Electrophoresis was used to determine the surface charge density of the rat erythrocyte membrane. It was shown that the process of aging was accompanied by a decrease in TAS and in the total amount of phospholipids as well as by enhancement of lipid peroxidation and increase in surface charge density of erythrocyte membrane. Ethanol administration caused, in term, decrease in TAS and increase in the level of all phospholipids and lipid peroxidation products. Ethanol as well significantly enhanced changes in surface charge density of erythrocyte membrane. The ingestion of green tea partially prevented decrease in erythrocyte antioxidant abilities observed during aging and ethanol intoxication. Moreover, long-term drinking of green tea protects the structure of the erythrocytes membrane disturbed during aging process and/or chronic ethanol intoxication.

Changes in electric charge and phospholipids composition in erythrocyte membrane of ethanol--poisoned rats after administration of teas.

Erythrocyte membrane components and electric charge can be modified by ethanol and by compounds formed as a result of its metabolism, particularly be reactive oxygen species (ROS). The effects of ROS can be neutralized by administering preparations with antioxidant properties. The natural preparations of this kind are teas. For this reason, it has been the purpose of this work to determine effect of green and black tea on membrane electric charge and phospholipids composition of an erythrocyte membrane from rats intoxicated with ethanol. Electrophoresis technique and HPLC have been applied to the above-mentioned studies. Ethanol administration caused an increase in erythrocyte surface charge density and phospholipids composition in the membrane. Administration of green and black tea with ethanol to the rats partially neutralizes the changes provoked by ethanol, and the action of green tea was stronger than that of black tea.

Changes in physico-chemical properties of human large intestine tumour cells membrane.

Tumour cells produce and excrete to blood many substances which are present in the cell itself in trace amounts only. Our work has been aimed at the determination of changes in electric charge and in phospholipid composition of large intestine normal mucosa and colorectal cancer cells. Surface charge density of tumour unaffected mucosa and of tissue sections from tumours, was measured by electrophoresis. The measurements were carried out at various pH of solution. Membrane isoelectric point was determined by measuring its electric charge in function of pH as well as total positive charge at low pH and total negative charge at high pH. Qualitative and quantitative composition of phospholipids in the membrane was determined by HPLC. Four phospholipid classes were identified: PI, PS, PE and PC and their surface concentrations were determined. The electric charge calculated from phospholipid concentrations is by three orders of magnitude higher than that determined electrophoretically. It indicates that the groups present in the membrane surface are involved in equilibria in which the charge is neutralized. The electric charge calculated from phospholipid concentrations is by three orders of magnitude higher than that determined electrophoretically. It indicates that the groups present in the membrane surface are involved in equilibria in which the charge is neutralized. Tumour changes provoke an increase in surface charge density of large intestine membrane, whereas the content of individual phospholipids increased or decreased depending on a patient.