title: Palmitate induces ER stress and autophagy in H9c2 cells: implications for apoptosis and adiponectin resistance.
abstract: The association between obesity and heart failure is well documented and recent studies have indicated that understanding the physiological role of autophagy will be of great significance. Cardiomyocyte apoptosis is one component of cardiac remodeling which leads to heart failure and in this study we used palmitate-treated H9c2 cells as an in vitro model of lipotoxicity to investigate the role of autophagy in cell death. Temporal analysis revealed that palmitate (100 μM) treatment induced a gradual increase of intracellular lipid accumulation as well as apoptotic cell death. Palmitate induced autophagic flux, determined via increased LC3-II formation and p62 degradation as well as by detecting reduced colocalization of GFP with RFP in cells overexpressing tandem fluorescent GFP/RFP-LC3. The increased level of autophagy indicated by these measures were confirmed using transmission electron microscopy (TEM). Upon inhibiting autophagy using bafilomycin we observed an increased level of palmitate-induced cell death assessed by Annexin V/PI staining, detection of active caspase-3 and MTT cell viability assay. Interestingly, using TEM and p-PERK or p-eIF2α detection we observed increased endoplasmic reticulum (ER) stress in response to palmitate. Autophagy was induced as an adaptive response against ER stress since it was sensitive to ER stress inhibition. Palmitate-induced ER stress also induced adiponectin resistance, assessed via AMPK phosphorylation, via reducing APPL1 expression. This effect was independent of palmitate-induced autophagy. In summary, our data indicate that palmitate induces autophagy subsequent to ER stress and that this confers a prosurvival effect against lipotoxicity-induced cell death. Palmitate-induced ER stress also led to adiponecin resistance.
authors.affiliation: Department of Biology, York University, Toronto, Canada.
authors.forename: Min
authors.initials: M
authors.lastname: Park
authors.forename: Anna
authors.initials: A
authors.lastname: Sabetski
authors.forename: Yee
authors.initials: Y
authors.lastname: Kwan Chan
authors.forename: Subat
authors.initials: S
authors.lastname: Turdi
authors.forename: Gary
authors.initials: G
authors.lastname: Sweeney
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000096:biosynthesis; D052242:Adiponectin / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects; D006333:Heart Failure / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D010168:Palmitates / Q000008:administration & dosage*
chemical_list: C464801:APPL1 protein, human; D048868:Adaptor Proteins, Signal Transducing; D052242:Adiponectin; D010168:Palmitates