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Peroxisome proliferator-activated receptors (PPAR`s) serve as lipid sensors and when activated modify gene expression of proteins highly involved in the regulation of fatty acid metabolism. Recently, the accumulation of lipids in liver was shown to be depended on the excessive protein-mediated transmembrane transport of long chain fatty acids (LCFAs). The aim of the present study was to determine the in vivo effects of PPAR and activation at two levels: 1) on the expression of fatty acid transporters, 2) on the content and fatty acids saturation status of lipids in rats liver. PPAR agonist (WY 14,643) treatment upregulated the liver expression of FAT/CD36 (+20%, p<0.05) and did not significantly affect the content of FABPpm and FATP-1. Accordingly there was a significant increase in the content of phospholipid (+12%, p<0.05), diacylglycerol (+65%, p<0.05) and triacylglycerol (+46%, p<0.05) fractions followed PPAR activation. In contrast, pioglitazone (PPAR agonist) had no effect on the content of fatty acid transporters (FAT/CD36, FABPpm and FATP-1) as well as the content of liver lipid fractions with the exception for triacylglycerols, which have been reduced significantly (-89%, p<0.05). These findings suggest that in vivo PPAR and PPAR activation exert different effects on both the expression of fatty acid transporters and lipid content in rat’s liver.
The fraction of free fatty acids (FFA) is present in skeletal muscles. However, there is almost no data regarding regulation in the content of this intramuscular lipid pool. We took advantage of the isolated muscle preparation to examine whether: a) increasing exogenous concentration of FFA (500µM or 700µM, 30min) b) insulin (10.00 I.U./L, 30min), c) adrenalin (4.4 nM, 30min), or d) contractions (200ms, tetani, 1Hz, 30min), affect the FFA content inside myocytes. Incubation of soleus (S) and extensor digitorum longus (EDL) with increasing concentrations of exogenous FFA (from 500µM to 700µM) resulted in an increase in the total FFA fraction in both muscles studied (by 280.2% and 259.1%, respectively). In contracting muscles FFA pool was significantly reduced both in S (by 73.1%) and in EDL (by 31.1%). Neither stimulation by adrenalin nor insulin affected the total content of FFA fraction in the muscles examined. We conclude that a) increased availability of exogenous FFA at the sarcolemma level results in an increase in the size of intramuscular FFA fraction b) the intracellular FFA fraction is utilized by contracting muscles with regard to the fiber composition and to a greater extent in more oxidative muscles, c) FFA fraction remains stable upon stimulation by insulin or adrenalin.
IL-6 is a biologically active substance and is thought to contribute to the development of obesity. Recent findings suggest that susceptibility to intracellular lipid accumulation is to a large extent determined by changes in the expression of fatty acid transporters such as FAT/CD36, FABPpm and FATP-1. The aim of the present study was to determine the effect of IL-6 deficiency on the expression of fatty acid transporters, as well as, assess the concomitant changes in intracellular lipids. We found that Il-6 deficiency upregulated the myocardial expression of FAT/CD36 (+40%) and did not significantly affect the content of FABPpm and FATP-1 (+15% and +5% respectively). Although no change in the intramyocardial total lipid content was noted, there was a significant increase in the intracellular content of both free fatty acid (FFA), diacylglicerol (DG) and ceramide fractions (+45%, +37% and +48%, respectively) in hearts from IL-6 -/- mice. A trend for IL-6 deficiency to increase in saturated FA species in these fractions was also observed (+8%, +12% and +10%, respectively). In contrast, IL-6 deficiency has no effect on the content of monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) species in each intramyocardial lipid fractions examinated. These findings suggest that IL-6 deficiency results in 1) upregulation of myocardial content of FAT/CD36, 2) the increase in the content of biologically active lipid pools (FFA, DG and ceramide). This lipid accumulation with concomitant trend for increase in the saturation status of these lipid fractions may, at least in part, provide a factor related to the development of intramyocardial lipotoxicity, observed in obese individuals.
Ceramide (Cer) and sphingomyelin (SM) are members of sphingolipid (SL) family. Their concentrationsin the brain undergo substantial changes in many pathologies. They are also important players in diabetes-linked brain dysfunctions, in which increased content of ceramides can be toxic to neurons. The aim of the study was to evaluate selected parameters of sphingolipids and insulin pathways in prefrontal cortex (PC) and hippocampus (H) of rats with experimentally induced hyperglycemia. STZ-rat model of type 1 diabetes and high fat diet model of insulin resistance were used. Analyses of studied parameters were performed by GLC, IHC and Elisa. We found the augmented levels of ceramides in H and PC and only minor in striatum and cerebellum of rats with STZ-induced diabetes. Similar expressions of Cer were confirmed by IHC. Myriocin, an inhibitor of an enzyme of ceramide de novo synthesis pathway, reduced ceramide generation in hyperglycemic brains, particularly in PC, which was reflected in altered Cer synthase activities. In addition, we reported the fluctuations in sphingomyelin levels in investigated structures. The level of insulin did not change in H and PC of STZ-treated rats. An expression of insulin receptor and its phosphorylated form decreased in both structures, but was restored after myriocin administration. Similarly, Akt and phosphorylated Akt changed in these structures suggesting important role of de novo Cer synthesis in intracellular pathways of insulin. In the rat model of high fat diet, which leads to insulin resistance, the sphingolipid pattern (Cer and SM) was altered in H and PC as well. Metformin, the drug of choice in diabetes type 2, influenced the content of the above SLs in these structures, suggesting the additional central activity of antidiabetic treatment. We conclude that ceramide and SM may be important mediators of diabetes- accompanied brain dysfunction.
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