Cerebral Phosphoinositide, Triacylglycerol, and Energy Metabolism in Reversible Ischemia: Origin and Fate of Free Fatty Acids

Abstract: Levels of phosphatidylinositol 4,5‐bisphosphate (PIP 2 ), phosphatidylinositol 4‐phosphate (PIP), phosphatidylinositol (PI), phosphatidic acid, diacylglycerol (DAG), triacylglycerol (TAG), and free fatty acids (FFAs), as well as their fatty acid composition, were determined in rat forebrain during ischemia and postischemic recirculation. Cerebral energy state and electroencephalograms (EEGs) were also studied. Fifteen minutes of ischemia resulted in a decrease in PIP 2 and PIP contents but not in PI content, concurrent with an enlargement of the FFA and DAG pools. The latter were enriched in stearate and arachidonate. Prolongation of ischemia did not produce further changes in content of any of the inositol phosphalipids, but the increase in levels of FFAs and DAG continued. At the end of 45 min of ischemia, levels of both PIP 2 and PIP decreased by 45–50%, and the total phosphoinositide content (PIP 2 + PIP + PI) decreased by 21%, whereas levels of FFAs and DAG increased to 14– and 3.6‐fold of control levels, respectively. During ischemia, the TAG‐palmitate level decreased, but the TAG‐arachidonate level increased; the tissue energy state deteriorated severely; and the EEG was suppressed. A 30‐min recirculation period after 15 or 45 min of ischemia led to increases in PIP 2 , PIP, and total phosphoinositide contents, whereas levels of FFAs and DAG promptly decreased toward control values. The TAG‐arachidonate level peaked and the TAG‐palmitate level returned to a low control value during early recirculation. The ischemic changes in tissue lipids were completely reversed within 3 h of recirculation after both periods of ischemia. Adenylates were fully phosphorylated with as little as 30 min of reflow. The EEG activity partially recovered during reflow after 15 min of ischemia, whereas it remained depressed after prolonged ischemia. Thus, phosphodiesteric cleavage of PIP 2 and PIP followed by deacylation of DAG is likely to contribute to the production of FFAs in early ischemia. Deacylation of undetermined lipids plays a role for the increment in levels of FFAs in the later period of ischemia. The rapid postischemic increase in levels of PIP 2 and PIP indicates active synthesis not only from existing PI, but probably also by means of accumulated FFAs and DAG. These results indicate that the impaired resynthesis of inositol phospholipids cannot be a cause of the poor EEG activity after prolonged ischemia. Degradation and resynthesis of polyphosphoinositides and formation of TAG‐arachidonate may be important for modulation of free arachidonic acid levels in the brain during temporary ischemia.