dinoprost and Carbon-Tetrachloride-Poisoning

Prostaglandin F2alpha (PGF2alpha), a foremost stable vasoactive cyclooxygenase (COX)-catalyzed prostaglandin, regulates a number of key physiological functions such as luteolysis, ovarian function, luteal maintenance of pregnancy, and parturition as a constitutive part of ongoing reproductive processes of the body. It has recently been implicated in the regulation of intricate pathophysiological processes, such as acute and chronic inflammation, cardiovascular and rheumatic diseases. Since the discovery of a second isoform of COXs, it has been shown that PGF2alpha can be formed in vivo from arachidonic acid through both isoforms of COXs, namely cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). Being synthesized in various parts of the body, it metabolizes instantly to a number of rather inactive metabolites mainly in the lungs, liver, kidney, and efficiently excretes into the urine. 15-Keto-dihydro-PGF2alpha, a major stable metabolite of PGF2alpha that reflects in vivo PGF2alpha biosynthesis, is found in larger quantities than its parent compound in the circulation and urine in basal physiological conditions, with short-lived pulses during luteolysis, induced termination of pregnancy and parturition, and is increased in tissues and various body fluids during acute, sub-chronic, and severe chronic inflammation. Further, the close relationship of PGF2alpha with a number of risk factors for atherosclerosis indicates its major role in inflammation pathology. This review addresses multiple aspects of PGF2alpha in addition to its emerging role in physiology to inflammation.

Topics: Animals; Atherosclerosis; Body Fluids; Carbon Tetrachloride Poisoning; Carotid Arteries; Cyclooxygenase 1; Cyclooxygenase 2; Diabetes Complications; Dinoprost; Female; Humans; Inflammation; Linoleic Acids, Conjugated; Luteolysis; Myocardial Reperfusion Injury; Obesity; Pregnancy; Risk Factors; Shock, Septic; Smoking

The effect of 15-Mt-PGF2 alpha on CCl4-induced injury of primary cultured hepatocytes was studied. 15-Mt-PGF2 alpha treatment (2 mg/L) significantly decreased CCl4 (10 mmol/L)-induced damages of primary cultured rat heptocytes as indicated by decreases GPT and GOT leakage and LPO production. 15-Mt-PGF2 alpha significantly promoted 3H-uridine incorporation into RNA and [3H]-thymidine incorporation into DNA of the rat hepatocytes. Cytopathology study showed that 15-Mt-PGF2 alpha attenuated damages of mitochondria, endoplasmic reticulum and ribosome caused by CCl4. 15-Mt-PGF2 alpha appeared to maintain the stability of rat hepatocytes by inhibiting lipid peroxidation. These results indicated that 15-Mt-PGF2 alpha has notable protective effect on primary cultured rat hepatocytes against CCl4-induced damage by reducing lipid peroxidation and promoting synthesis of RNA and DNA.

Topics: Animals; Carbon Tetrachloride Poisoning; Cells, Cultured; Chemical and Drug Induced Liver Injury; Dinoprost; Female; Male; Rats; Rats, Sprague-Dawley

Although numerous methods have been developed for the detection of lipid peroxidation, it is generally recognized that most of these lack specificity and/or sensitivity, particularly when applied to in vivo situations. We have reported recently that a series of prostaglandin F2-like compounds, termed F2-isoprostanes, are formed in vivo from the free radical catalyzed peroxidation of arachidonic acid and appear to be a useful marker of oxidant stress. Because of formation of other products of lipid peroxidation, such as alkanes and malondialdehyde (MDA), are affected by oxygen tension, which may influence their usefulness as markers of oxidant stress, we carried out a systematic study of the generation of F2-isoprostanes at various oxygen concentrations and compared these changes with the generation of MDA. The disappearance of the F2-isoprostane precursor, arachidonic acid, was used as a reference measure. Rat liver microsomes were peroxidized using an iron-ascorbate system. The incubations were carried out in sealed flasks at 37 degrees under N2 and various concentrations of O2 up to 100%. F2-isoprostanes were quantified by mass spectrometry and MDA by the thiobarbituric acid reaction. Microsomal fatty acids were measured by gas chromatography. Both MDA and F2-isoprostane formation increased in a time-dependent manner up to 15 min. Their formation correlated with a loss of polyunsaturated fatty acid and with an increase in O2 tension up to 21% O2. At oxygen tensions above 21%, MDA generation continued to increase, while F2-isoprostane generation and arachidonic acid loss did not. Levels of MDA and F2-isoprostanes increased a maximum of 65 and 9.4 times baseline values, respectively. These studies, therefore, define factors that influence the formation of F2-isoprostanes in an in vitro model of lipid peroxidation. Further, they demonstrate that higher O2 tensions do not block formation of F2-isoprostanes and validate their usefulness for assessing lipid peroxidation under high, as well as low, oxygen tension.

Topics: Animals; Arachidonic Acid; Carbon Tetrachloride Poisoning; Dinoprost; Dose-Response Relationship, Drug; Lipid Peroxidation; Male; Malondialdehyde; Microsomes, Liver; Oxygen; Rats; Rats, Sprague-Dawley

We recently reported the discovery of a series of bioactive prostaglandin F2-like compounds (F2-isoprostanes) that are produced in vivo by free radical-catalyzed peroxidation of arachidonic acid independent of the cyclooxygenase enzyme. Inasmuch as phospholipids readily undergo peroxidation, we examined the possibility that F2-isoprostanes may be formed in situ on phospholipids. Initial support for this hypothesis was obtained by the finding that levels of free F2-isoprostanes measured after hydrolysis of lipids extracted from livers of rats treated with CCl4 to induce lipid peroxidation were more than 100-fold higher than levels in untreated animals. Further, increased levels of lipid-associated F2-isoprostanes in livers of CCl4-treated rats preceded the appearance of free compounds in the circulation, suggesting that the free compounds arose from hydrolysis of peroxidized lipids. This concept was supported by demonstrating that free F2-isoprostanes were released after incubation of lipid extracts with bee venom phospholipase A2 in vitro. When these lipid extracts were analyzed by HPLC, fractions that yielded large quantities of free F2-isoprostanes after hydrolysis eluted at a much more polar retention volume than nonoxidized phosphatidylcholine. Analysis of these polar lipids by fast atom bombardment mass spectrometry established that they were F2-isoprostane-containing species of phosphatidylcholine. Thus, unlike cyclooxygenase-derived prostanoids, F2-isoprostanes are initially formed in situ on phospholipids, from which they are subsequently released preformed, presumably by phospholipases. Molecular modeling of F2-isoprostane-containing phospholipids reveals them to be remarkably distorted molecules. Thus, the formation of these phospholipid species in lipid bilayers may contribute in an important way to alterations in fluidity and integrity of cellular membranes, well-known sequelae of oxidant injury.

Topics: Animals; Carbon Tetrachloride; Carbon Tetrachloride Poisoning; Chromatography, High Pressure Liquid; Dinoprost; Free Radicals; Gas Chromatography-Mass Spectrometry; Lipid Peroxidation; Liver; Microsomes, Liver; Models, Molecular; Molecular Conformation; Phospholipids; Rats; Spectrometry, Mass, Fast Atom Bombardment