Palmitic and stearic fatty acids induce caspase‐dependent and ‐independent cell death in nerve growth factor differentiated PC12 cells

Abstract

Apoptotic cell death has been proposed to play a role in the neuronal loss observed following traumatic injury in the CNS and PNS. The present study uses an in vitro tissue culture model to investigate whether free fatty acids (FFAs), at concentrations comparable to those found following traumatic brain injury, trigger cell death. Nerve growth factor (NGF)-differentiated PC12 cells exposed to oleic and arachidonic acids (2 : 1 ratio FFA/BSA) showed normal cell survival. However, when cells were exposed to stearic and palmitic acids, there was a dramatic loss of cell viability after 24 h of treatment. The cell death induced by stearic acid and palmitic acid was apoptotic as assessed by morphological analysis, and activation of caspase-8 and caspase-3-like activities.Western blotting showed that differentiated PC12 cells exposed to stearic and palmitic acids exhibited the signature apoptotic cleavage fragment of poly (ADP-ribose) polymerase (PARP). Interestingly, blockade of caspase activities with the pan-caspase inhibitor z-VAD-fmk failed to prevent the cell death observed induced by palmitic or stearic acid. RT-PCR and RNA blot experiments showed an up-regulation of the Fas receptor and ligand mRNA. These findings are consistent with our hypothesis that FFAs may play a role in the cell death associated with trauma in the CNS and PNS.

The Effect of Poloxamer-188 on Neuronal Cell Recovery from Mechanical Injury

“…These cells have been well characterized and have become a model for many studies related to identification and characterization of neuronal signaling pathways (Heneka et al, 1998), to study cell death (Dubreuil et al, 2003;Kitazawa et al, 2004;Ulloth et al, 2003;Yang et al, 2004), especially apoptosis, in neuronal cells and cytoskeletal structure and function (Kobayashi and Mundel, 1998;Nakayama et al, 2001;Okabe and Hirokawa, 1988;Tahir et al, 1992). Although no other types of cells were examined in the present study, cell viability studies and onset rate dependency of traumatic injury produced on cells with CCSD were similar to previous studies conducted with human derived neuronal cells (Ellis et al, 1995;LaPlaca and Thibault, 1998).…”

Free Fatty Acids Inhibit Serum Deprivation-induced Apoptosis through GPR120 in a Murine Enteroendocrine Cell Line STC-1

“…Saturated FFAs have been reported to have toxicity on various types of cells, such as breast cancer MDA-MB-231 cells (27,28), pheochromocytoma PC12 cells (29), Chinese hamster ovary cells (30), pancreatic beta cells (31), rat testicular Leydig cells (32), and rat neonatal cardiomyocytes (33). However, studies on FFA-induced effects are limited, and the mechanism of such effects remains largely unknown.…”

In situ assay of fatty acid β-oxidation by metabolite profiling following permeabilization of cell membranes

“…Although albumin is frequently used, methyl-␤ -cyclodextrin (MBCD) is an alternative vehicle for the solubilization of FAs ( 25,26 ). We found that MBCD applied as described previously ( 25 ) led to an improved reconstitution of defi ned acylcarnitine standards in solution compared with albumin.…”