Hipolito TRUE posted an update 2 weeks, 1 day ago
Fatty acids can regulate flux through energy metabolic pathways, and may thereby regulate cell survival. The LY2109761 inquirer survival and proliferation of cells with high glycolytic rates tends to be positively correlated with glycolysis. In other cell types a process referred to as the glucosefatty acid cycle, or the Randle cycle, has been observed, where increased fatty acid oxidation can inhibit glucose oxidation and glycolysis. However, in the heart it has frequently been reported that elevating fatty acid oxidation results in uncoupling of glycolysis from glucose oxidation, due to a greater inhibition of glucose oxidation than glycolysis. In agreement with this, inhibiting fatty acid oxidation via MCD inhibition results in pulmonary artery smooth muscle cell apoptosis and decreased proliferation. This is probably detrimental to these cells because decreasing palmitate oxidation likely results in an improved coupling of glycolysis to glucose oxidation. Therefore, this link between fatty acid oxidation and glucose metabolism could explain why fatty acid oxidation seems to regulate cell proliferation and survival. An alternative explanation for the effects of fatty acids on cell survival is that fatty acid oxidation could be beneficial independent of its effects on glycolysis. It has been suggested that under conditions where glycolysis is reduced fatty acid oxidation can be used by cancer cells for energy production. We therefore decided to determine whether fatty acids inhibit human BMMSC glucose metabolism and induce BMMSC death via modulation of glucose and fatty acid energy metabolism. Acute exposure to palmitate and/or oleate did not affect glycolysis or fatty acid oxidation rates. However, combined acute exposure to palmitate and oleate did reduce glucose oxidation. These results indicate that the Randle Cycle exists at least to some extent in human BMMSCs. Following chronic treatment with palmitate and/or oleate we observed that only palmitate exposure reduced palmitate oxidation rates. Interestingly, combined treatment with oleate, which prevented palmitate-induced death, prevented this reduction in palmitate oxidation. Neither palmitate or oleate affected the expression of proteins involved in oxidative metabolism or glycolysis that we measured. These palmitate oxidation results agree with a previous report showing that 20 hr exposure of neonatal cardiac myocytes to palmitate induced apoptosis and decreased palmitate oxidation rates. This suggests that palmitate induces BMMSC death via inhibition of palmitate oxidation and that oleate is protective because it prevents palmitate oxidation from decreasing. This is supported by a previous study in BMMSCs in which AICAR prevented palmitate-induced death. However, we did not observe a change in phosphorylation of ACC, an indicator of AMPK activity and an important pathway by which AMPK increases fatty acid oxidation. This may not actually be that surprising, since AMPK activation can decrease proliferation but as we show here oleate protects against the drop in proliferation induced by palmitate. It is still a possibility, however, that a reduction in glycolysis may be involved in palmitate-induced BMMSC death, but changes in glycolysis that occur in response to 24 hr treatment with palmitate and/or oleate were masked by switching all groups to the same buffer during the measurement of glycolysis rates. Oleate had a dramatic effect of preventing palmitate-induced BMMSC death. This may have occurred secondary to inhibiting palmitate uptake. Acutely, oleate and palmitate reduced each other’s uptake. However, after 24 hr of exposure to palmitate and/or oleate, palmitate uptake was not different between groups. It is still possible, however, that oleate did in fact reduce palmitate uptake at 24 hr but it was an acute effect and therefore was not measured. Therefore, oleate may be at least partially protecting against palmitate-induced cell death by reducing intracellular palmitate levels by decreasing palmitate uptake. Another potential mechanism for palmitate-induced cell death in the BMMSCs is the potential involvement of ceramides. Elevated levels of ceramides are able to induce death in a number of different cell types. The fact that saturated fatty acids, which are ceramide substrates, induced BMMSC death while oleate, an unsaturated fatty acid which is not a ceramide substrate, does not induce BMMSC death suggests that ceramides could be involved in saturated fatty acid-induced BMMSC death. In addition, chronic exposure to palmitate reduces fatty acid oxidation, which could result in a redirection of palmitate into ceramides.