Fish oil increases mitochondrial phospholipid unsaturation, upregulating reactive oxygen species and apoptosis in rat colonocytes.
We have shown that a combination of fish oil (high in n-3 fatty acids) with the butyrate-producing fiber pectin, upregulates apoptosis in colon cells exposed to the carcinogen azoxymethane, protecting against colon tumor development. We now hypothesize that n-3 fatty acids prime the colonocytes such that butyrate can initiate apoptosis. To test this, 30 Sprague-Dawley rats were provided with diets differing in the fatty acid composition (corn oil, fish oil or a purified fatty acid ethyl ester diet). Intact colon crypts were exposed ex vivo to butyrate, and analyzed for reactive oxygen species (ROS), mitochondrial membrane potential (MMP), translocation of cytochrome C to the cytosol, and caspase-3 activity (early events in apoptosis). The fatty acid composition of the three major mitochondrial phospholipids was also determined, and an unsaturation index calculated. The unsaturation index in cardiolipin was correlated with ROS levels (R = 0.99; P = 0.02). When colon crypts from fish oil and FAEE-fed rats were exposed to butyrate, MMP decreased (P = 0.041); and translocation of cytochrome C to the cytosol (P = 0.037) and caspase-3 activation increased (P = 0.032). The data suggest that fish oil may prime the colonocytes for butyrate-induced apoptosis by enhancing the unsaturation of mitochondrial phospholipids, especially cardiolipin, resulting in an increase in ROS and initiating apoptotic cascade.
Hong
Mee Young
MY
Faculty of Nutrition, Texas A&M University, College Station, TX 77843-2471, USA.
Chapkin
Robert S
RS
Barhoumi
Rola
R
Burghardt
Robert C
RC
Turner
Nancy D
ND
Henderson
Cara E
CE
Sanders
Lisa M
LM
Fan
Yang-Yi
YY
Davidson
Laurie A
LA
Murphy
Mary E
ME
Spinka
Christine M
CM
Carroll
Raymond J
RJ
Lupton
Joanne R
JR
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002087:Butyrates / Q000494:pharmacology; D002308:Cardiolipins / Q000737:chemistry / Q000378:metabolism; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D003106:Colon / Q000166:cytology* / Q000378:metabolism; D003314:Corn Oil / Q000494:pharmacology; D003574:Cytochrome c Group / Q000378:metabolism; D004042:Dietary Fats, Unsaturated / Q000494:pharmacology; D004789:Enzyme Activation; D004952:Esters / Q000494:pharmacology; D005227:Fatty Acids / Q000494:pharmacology; D005395:Fish Oils / Q000494:pharmacology*; D007413:Intestinal Mucosa / Q000166:cytology* / Q000378:metabolism; D007425:Intracellular Membranes / Q000187:drug effects* / Q000378:metabolism; D015227:Lipid Peroxidation; D008297:Male; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*
D002087:Butyrates; D002308:Cardiolipins; D003574:Cytochrome c Group; D004042:Dietary Fats, Unsaturated; D004952:Esters; D005227:Fatty Acids; D005395:Fish Oils; D008563:Membrane Lipids; D010743:Phospholipids; D017382:Reactive Oxygen Species; D003314:Corn Oil; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases