The significance of base-exchange reactions in lipid metabolism.
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Biomedical subjects
Publications and source records attributed to G Trovarelli.
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It has been previously demonstrated that spontaneously hypertensive adult rats (SHR) develop severe hypertension and cerebrovascular lesions on drinking 1% NaCl from weaning and that the phospholipid metabolism in the whole brain is actively altered in these lesioned animals (SHR-NaCl) as compared to SHRs which drink only water and show only sporadic cerebrovascular lesions. We have now assayed the incorporation of labelled choline, ethanolamine, glycerol and arachidonic acid into the phospholipids from the cortex and hippocampus of SHR-water and SHR-NaCl at different time intervals from injection into the lateral ventricle of the brain. A noticeable decrease of both choline and arachidonate specific activity (SA) in the phospholipids was found in the cortex and hippocampus (where the effect is most evident) from SHR-NaCl. Based on the literature and the data obtained, we suggest that in SHR-NaCl brain areas a release of choline and fatty acid also occurs from choline glycerophospholipids as a consequence of the cerebrovascular lesions caused by NaCl treatment. Even if a relatively minor loss of the amount of the lipids studied is evident from our results as compared to their entire pool, this change may be quite important if it causes a modification of the lipidic bilayer in excitable membranes. In a parallel group of SHR-NaCl animals, treated with the nootropic drug oxiracetam, we observed that the metabolic utilization of the precursors was completely restored. These experimental data favour the hypothesis that oxiracetam is effective in stimulating the phospholipid metabolism rate at levels even higher than those of the SHR-water animals.
The linoleic acid metabolism was examined in the brain cortex of 4 month-old and 24 month-old rats. After the injection of [1-14C]-linoleate into the lateral ventricle of the brain the animals were sacrificed at 1,3 and 6 hours from the injection. The linoleate (18:2) incorporation into lipids, the presence of fatty acid peroxidation products, as well as the 18:2 transformation into elongated and desaturated derivatives were determined. Both an age-related reduction in linoleate incorporation rate into glycerophospholipids and a decrease in fatty acid turnover were found. Furthermore, in glycerophospholipids from 24 month-old rat brain cortex a higher level of hydroperoxide derivative of linoleate was found as compared to 4 month-old animals, and this damaged fatty acid is eliminated more slowly in aged rats than in adults. Finally, unlike 4 month-old animals, a stimulation of the transformation rate of linoleate into desaturation (6,9,12-C18:3) and elongation (8,11,14,C20:3) products was found in 24 month-old rat brain cortex. On the contrary, as far as arachidonic acid (one of the most important end products of the mechanism of linoleate modification) is concerned, the differences between aged and control animals were small, making it quite difficult to attribute a physiological meaning to this phenomenon.
It has been demonstrated that spontaneously hypertensive adult rats (SHR) develop severe hypertension and cerebrovascular lesions on drinking 1% NaCl from weaning. Phospholipid metabolism is actively altered in these severely lesioned animals (SHR-NaCl) as compared to SHRs which drink only water and showed only sporadic cerebrovascular lesions. We have tested the incorporation of water soluble phospholipid precursors into the corresponding phospholipid from different brain areas, by injecting either a mixture of labeled glycerol and choline or glycerol and ethanolamine into the lateral ventricle of the brain of adult (4 months old) and senescent (12 months old) SHR-NaCl. The results were compared to those obtained from 4 and 12 months old Wistar normotensive rats. When adult normotensive rats were compared with adult hypertensive rats (4-SHR-NaCl) incorporation was found to decrease in some areas according to the precursors injected. Similar results were obtained from 12 month old normotensive Wistar rats that, however, showed a decrease in phospholipid biosynthesis in all the area tested. Interestingly, no significant differences of incorporation rate were found between 12 month old normotensive and 12 month old hypertensive rats.
Brain ischemia was produced in gerbils by contemporary occlusion of both carotid arteries. Definite changes of the energy state in brain demonstrated that carotid occlusion was effective. At short time intervals from occlusion the free fatty acid content, their distribution, and their concentration and specific activity in arachidonate were determined in brain. A noticeable increase of the arachidonate pool and that of other free fatty acids was detected at very early times from occlusion. Specific activity by arachidonate increased after 30-60 seconds from ligation. By examining arachidonate distribution and specific activity in neutral and polar lipids of brain, it is concluded that phosphatidylcholine and phosphatidylinositol represent the more important source for the release of arachidonate during ischemia. Enzymic-mediated phenomena produced free arachidonate from lipids by a mechanism yielding diglycerides further transformed into fatty acids and by lipid degradation through phospholipase A activity.
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The capillary endothelium of cerebral microvessels plays an important role in homeostasis within the central nervous system. The flux of fluids and solutes takes place through the lipid matrix of plasma membranes and the maintenance of their structural composition is necessary for cell membrane permeability and the cellular transport systems. Few studies have been carried out to clarify the relationships between brain microvessel lipid metabolism and the role of this metabolism. We have studied a particular aspect of the phospholipid metabolism, i.e. their hydrosoluble head group exchange with free choline, ethanolamine or serine, in capillaries from brains of both 4 month- old and 24 month-old rats. The results obtained indicate that microvessels possess the biochemical machinery of base-exchange reaction, whose activity rate appears unaffected by age.
The calcium-stimulated incorporation of ethanolamine, L-serine and choline into rabbit synaptosomal phospholipids in vitro has been investigated. The synaptosomal membranes were prelabelled in vitro in their choline-, ethanolamine- or serine-phosphoglycerides by base-exchange and then chasing experiments were performed by displacing the lipid-bound base by ethanolamine, choline of L-serine labelled with a different isotope. The results indicate that membrane phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine are substrates for the exchange with all the three mentioned bases. A very small phospholipid pool (0.5-2% of the total available pool) is active in the calcium-dependent exchange between membrane phosphatidylcholine or phosphatidylethanolamine and free bases, whereas the pool of exchanging phosphatidylserine molecules is sensibly larger (2-9%). The possible physiological importance of these small phospholipid pools, involved in base-exchange reaction at brain membrane level, is discussed.