Mitochondrial dysfunction during in vitro hepatocyte steatosis is reversed by omega-3 fatty acid-induced up-regulation of mitofusin 2.
We examined the effects and mechanisms of omega-3 polyunsaturated fatty acid (PUFA) administration on mitochondrial morphology and function in an in vitro steatotic hepatocyte model created using HepG2 cells. Reverse transcriptase polymerase chain reaction and Western blot analyses were performed to determine the expression levels of mitofusin 2 (Mfn2), and immunofluorescent MitoTracker Mitochondrion-Selective Probes were used to detect changes in mitochondrial morphology. Adenosine triphosphate (ATP) synthesis and reactive oxygen species (ROS) production were measured to assess mitochondrial function. Mitofusin 2 expression was significantly suppressed (P < .05), ATP levels were decreased (P < .05), ROS production was increased (P < .05), and the normal tubular network of mitochondria was fragmented into short rods or spheres. Model cells were incubated with eicosapentaenoic acid or docosahexaenoic acid at a final concentration of 50 μmol/L for 1 hour. Both eicosapentaenoic acid and docosahexaenoic acid increased the expression of Mfn2 (P < .01) and caused an increase in the length of mitochondrial tubules. The omega-3 PUFAs also increased the levels of ATP (P < .05) and decreased the ROS production (P < .05). However, these changes were not seen in Mfn2-depleted steatotic HepG2 cells, created by RNA interference before incubation with the omega-3 PUFAs. This study demonstrated that, in steatotic hepatocytes, omega-3 PUFAs may change mitochondrial morphology and have beneficial effects on recovery of mitochondrial function by increasing the expression of Mfn2.
Zhang
Yong
Y
Department of General Surgery, Union Hospital, Huazhong University of Science and Technology, China, 430022.
Jiang
Longchang
L
Hu
Wenjun
W
Zheng
Qichang
Q
Xiang
Wenpei
W
D000255:Adenosine Triphosphate / Q000378:metabolism; D001391:Azo Compounds; D015153:Blotting, Western; D002478:Cells, Cultured; D004396:Coloring Agents; D015525:Fatty Acids, Omega-3 / Q000627:therapeutic use*; D005234:Fatty Liver / Q000188:drug therapy* / Q000473:pathology*; D005470:Fluorometry; D020558:GTP Phosphohydrolases; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008565:Membrane Proteins / Q000096:biosynthesis*; D008856:Microscopy, Fluorescence; D008930:Mitochondria, Liver / Q000187:drug effects / Q000473:pathology* / Q000648:ultrastructure; D024101:Mitochondrial Proteins / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects
D001391:Azo Compounds; D004396:Coloring Agents; D015525:Fatty Acids, Omega-3; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D000255:Adenosine Triphosphate; D020558:GTP Phosphohydrolases; C470018:MFN2 protein, human; C011049:oil red O