کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5948304 1172378 2012 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Palmitate induces RIP1-dependent necrosis in RAW 264.7 cells
موضوعات مرتبط
علوم پزشکی و سلامت پزشکی و دندانپزشکی کاردیولوژی و پزشکی قلب و عروق
پیش نمایش صفحه اول مقاله
Palmitate induces RIP1-dependent necrosis in RAW 264.7 cells
چکیده انگلیسی

ObjectiveThe kinase receptor-interacting protein (RIP) 1, a serine/threonine protein kinase, is a key signaling molecule for necrosis. The possible involvement of RIP1 in palmitate-induced macrophage death and its underlying molecular mechanism was investigated in this study.MethodsCell viability was measured by an MTT reduction assay. The type of cell death was determined by staining with annexin V, propidium iodide (PI) and the APOPercentage dye, and by examining cell morphology using transmission electron microscopy. The down-regulation of RIP1 was performed by siRNA transfection. Intracellular reactive oxygen species (ROS) were measured by staining with H2DCF-DA.ResultsPalmitate largely induced necrosis in RAW 264.7 cells, whereas C2-ceramide induced apoptosis. Palmitate-induced necrosis was inhibited by Necrostatin-1, an inhibitor of RIP1, and by RIP1 siRNA transfection, whereas ordinary cell death was not inhibited by z-VAD-fmk. In addition, the presence of palmitate caused a significant increase in intracellular ROS levels compared to control cells. Pre-treatment with Tempol, a cell permeable ROS scavenger, and MnTBAP, an inhibitor of mitochondrial oxidative stress, protected cells from palmitate-induced cell death. Furthermore, the down-regulation of RIP1 by siRNA transfection significantly decreased palmitate-induced ROS generation compared to control cells.ConclusionThe findings reported herein indicate that palmitate induces necrotic cell death via RIP1-dependent ROS generation in RAW 264.7 cells. These findings may provide a new mechanism that explains the link between elevated levels of free fatty acids (FFAs) and macrophage death.

► Palmitate-induced necrosis in RAW 264.7 cells. ► Palmitate-induced necrosis was inhibited by Necrostatin-1 and by RIP1 silencing. ► Palmitate caused a significant increase in intracellular ROS levels. ► The RIP1 silencing decreased palmitate-induced ROS generation.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Atherosclerosis - Volume 225, Issue 2, December 2012, Pages 315-321
نویسندگان
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