Abstract
The purpose of this study was to explore the mechanism of by which docosahexaenoic acid (DHA) inhibit the accumulation of adipose tissue lipid in grass carp (Ctenopharyngodon idella). We therefore designed two semi-purified diets, namely DHA-free (control) and DHA-supplemented, and fed them to grass carp (22.19 ± 1.76 g) for 3 and 6 weeks. DHA supplementation led to a significantly lower intraperitoneal fat index (IPFI) than that in the control group by reducing the number of adipocytes but significantly higher adipocyte size (P < 0.05). In the intraperitoneal adipose tissue, the DHA-fed group showed significantly higher peroxisome proliferator-activated receptor (PPAR)γ, CCAAT enhancer-binding protein (C/EBP)α, and sterol regulatory element-binding protein (SREBP)1c mRNA expression levels at both 3 and 6 weeks (P < 0.05). However, the ratio of the expression levels of B cell leukemia 2 (Bcl-2) and Bcl-2-associated X protein (Bax) was significantly lower in the DHA-fed group than in the control group (P < 0.05), and the protein expression levels of the apoptosis-related proteins caspase 3, caspase 8, and caspase 9 were also significantly higher (P < 0.05). Overall, although DHA promotes lipid synthesis, it is more likely that DHA could suppress the lipid accumulation in adipocytes of grass carp by inducing adipocyte apoptosis.
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Ahangar P, Sam MR, Nejati V, Habibian R (2016) Treatment of undifferentiated colorectal cancer cells with fish-oil derived docosahexaenoic acid triggers caspase-3 activation and apoptosis. Journal of Cancer Research & Therapeutics 12:798
SMR, Lazar MA (2000) Transcriptional control of adipogenesis. Annu Rev Nutr 20:535
Arner P, Spalding KL (2010) Fat cell turnover in humans. Biochem Biophys Res Commun 396(1):101–104
Aggoun Y (2007) Obesity, metabolic syndrome, and cardiovascular disease. Journal of Clinical Endocrinology & Metabolism 89:2595–2600
Arzel J, Lopez FXM, Métailler R, Stéphan G, Viau M, Gandemer G, Guillaume J (1994) Effect of dietary lipid on growth performance and body composition of brown trout (Salmo trutta ) reared in seawater. Aquaculture 123:361–375
Calder PC (2012) Mechanisms of action of (n-3) fatty acids. J Nutr 142:592S
Chaiyapechara S, Casten MT, Hardy RW, Dong FM (2003) Fish performance, fillet characteristics, and health assessment index of rainbow trout (Oncorhynchus mykiss) fed diets containing adequate and high concentrations of lipid and vitamin E. Aquaculture 219:715–738
Cowey CB, Cho CY (1993) Nutritional requirements of fish. Proc Nutr Soc 52:417
Dias J, Alvarez MJ, Diez A, Arzel J, Corraze G, Bautista JM, Kaushik SJ (1998) Regulation of hepatic lipogenesis by dietary protein/energy in juvenile European seabass (Dicentrarchus labrax). Aquaculture 161:169–186
Du ZY, Clouet P, Huang LM et al (2008) Utilization of different dietary lipid sources at high level in herbivorous grass carp (Ctenopharyngodon idella): mechanism related to hepatic fatty acid oxidation [J]. Aquac Nutr 14(1):77–92
Fajas L, Fruchart JC, Auwerx J (1998) Transcriptional control of adipogenesis. Curr Opin Cell Biol 10:165–173
Folch J, Lees M, Stanley GHS (1957) A simple method for the isolation and purification of total lipids from animal tissue. J Biol Chem 226:497–509
Frayn KN (2002) Adipose tissue as a buffer for daily lipid flux. Diabetologia 45:1201–1210
Frayn KN, Arner P, Ykijärvinen H (2006) Fatty acid metabolism in adipose tissue, muscle and liver in health and disease. Essays Biochem 42:89–103
Guo X, Liang XF, Fang L, Yuan X, Zhou Y, Zhang J, Li B (2015) Effects of dietary non-protein energy source levels on growth performance, body composition and lipid metabolism in herbivorous grass carp (Ctenopharyngodon idella Val.) Aquac Res 46:1197–1208
Hanada H, Morikawa K, Hirota K, Nonaka M, Umehara Y (2011) Induction of apoptosis and lipogenesis in human preadipocyte cell line by n-3 PUFAs. Cell Biol Int 35:51–59
Hernández-Cruz CM, Mesa-Rodríguez A, Betancor M, Haroun-Izquierdo A, Izquierdo M, Benítez-Santana T, Torrecillas S, Roo J (2015) Growth performance and gene expression in gilthead sea bream (Sparus aurata) fed microdiets with high docosahexaenoic acid and antioxidant levels. Aquac Nutr 21:881–891
Huang TS, Todorčević M, Ruyter B, Torstensen B (2010) Altered expression of CCAAT/enhancer binding protein and FABP11 genes during adipogenesis in vitro in Atlantic salmon (Salmo salar). Aquac Nutr 16:72–80
Jean OC, Lydia L, Cawood TJ, Anna K, Niamh N, Justin G, Aiden MC, Cliona OF, Donal OS (2010) The relationship of omental and subcutaneous adipocyte size to metabolic disease in severe obesity. PLoS One 5:e9997
Jeong S, Jing K, Kim N, Shin S, Kim S, Song KS, Heo JY, Park JH, Seo KS, Han J (2014) Docosahexaenoic acid-induced apoptosis is mediated by activation of mitogen-activated protein kinases in human cancer cells. BMC Cancer 14:481
Ji H, Cao YZ, Liu P, Su SS, Lin YQ, Cao FY, Oku H, Zhou JS, Ye YT (2009) Effect of dietary HUFA on the lipid metabolism in grass carp Ctenopharymgodon idellus. Acta Hydrobiol Sinica 33:881–889
Ji H, Li J, Liu P (2011) Regulation of growth performance and lipid metabolism by dietary n-3 highly unsaturated fatty acids in juvenile grass carp, Ctenopharyngodon idellus. Comparative Biochemistry & Physiology Part B 159:49–56
Jing K, Song KS, Shin S, Kim N, Jeong S, Oh HR, Park JH, Seo KS, Heo JY, Han J (2011) Docosahexaenoic acid induces autophagy through p53/AMPK/mTOR signaling and promotes apoptosis in human cancer cells harboring wild-type p53. Autophagy 7:1348–1358
Lee SM, Jeon IG, Lee JY (2002) Effects of digestible protein and lipid levels in practical diets on growth, protein utilization and body composition of juvenile rockfish ( Sebastes Schlegeli ). Aquaculture 211:227–239
Liu P, Ji H, Li C, Tian J, Wang Y, Yu P (2015) Ontogenetic development of adipose tissue in grass carp (Ctenopharyngodon idellus). Fish Physiol Biochem 41:1–12
Liu P, Li C, Huang J, Ji H (2014) Regulation of adipocytes lipolysis by n-3 HUFA in grass carp (Ctenopharyngodon idellus) in vitro and in vivo. Fish Physiol Biochem 40:1447–1460
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25:402–408
Lovell T, (1989) Nutrition and feeding of fish[M]. New York, Van Nostrand Reinhold
Ma JJ, Shao QJ, Xu ZR et al (2009) Effects of dietary n-3 HUFA on growth performance and lipid metabolism in juvenile black sea bream Sparus macrocephalus[J]. J Fish China 33(4):639–649
Metcalfe LC, Schmitz AL, (1961) A new method for saponification and methylation of fatty acids[J]. Anal Chem 33:363-372
Kanazawa A (1997) Effects of docosahexaenoic acid and phospholipids on stress tolerance of fish. Aquaculture 155:129–134
Mayer B, Oberbauer R (2003) Mitochondrial regulation of apoptosis [J]. Physiology 18(3):89–94
Notarnicola M, Messa C, Refolo MG, Tutino V, Miccolis A, Caruso MG (2011) Polyunsaturated fatty acids reduce fatty acid synthase and hydroxy-methyl-glutaryl CoA-reductase gene expression and promote apoptosis in HepG2 cell line. Lipids Health Dis 10:1
Osman OS, Selway JL, Stocker CJ, O’Dowd JF, Cawthorne MA, Arch JR, Jassim S, Langlands K (2013) A novel automated image analysis method for accurate adipocyte quantification. Adipocyte 2:160–164
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research 29:e45
Rosen ED, Hsu C-H, Wang X, Sakai S, Freeman MW, Gonzalez FJ, Spiegelman BM (2002) C/EBPα induces adipogenesis through PPARγ: a unified pathway. Genes Dev 16:22–26
Ruzickova J, Rossmeisl M, Prazak T, Flachs P, Sponarova J, Vecka M, Tvrzicka E, Bryhn M, Kopecky J (2004) Omega-3 PUFA of marine origin limit diet-induced obesity in mice by reducing cellularity of adipose tissue. Lipids 39:1177
Skender B, Hofmanová J, Slavík J, Jelínková I, Machala M, Moyer MP, Kozubík A, Hyršlová VA (2014) DHA-mediated enhancement of TRAIL-induced apoptosis in colon cancer cells is associated with engagement of mitochondria and specific alterations in sphingolipid metabolism. Biochim Biophys Acta 1841:1308–1317
Stowell SL, Iii DMG (1992) Effects of dietary pantethine and lipid levels on growth and body composition of channel catfish, Ictalurus punctatus. Aquaculture 108:177–188
Tian JJ, Lu RH, Ji H, Sun J, Li C, Liu P, Lei CX, Chen LQ, Du ZY (2015) Comparative analysis of the hepatopancreas transcriptome of grass carp (Ctenopharyngodon idellus) fed with lard oil and fish oil diets. Gene 565:192–200
Todorčević M, Hodson L (2016) The effect of marine derived n-3 fatty acids on adipose tissue metabolism and function. J Clin Med 5:3
Todorčević M, Kjaer MA, Djaković N, Vegusdal A, Torstensen BE, Ruyter B (2009) N-3 HUFAs affect fat deposition, susceptibility to oxidative stress, and apoptosis in Atlantic salmon visceral adipose tissue. Comparative Biochemistry & Physiology Part B Biochemistry & Molecular Biology 152:135–143
Takle H, McLeod A, Andersen O (2006) Cloning and characterization of the executioner caspases 3, 6, 7 and Hsp70 in hyperthermic Atlantic salmon (Salmo salar) embryos. Comp Biochem Physiol B: Biochem Mol Biol 144(2):188–198
Tsuzuki T, Kawakami Y, Nakagawa K, Miyazawa T (2006) Conjugated docosahexaenoic acid inhibits lipid accumulation in rats. Journal of Nutritional Biochemistry 17:518–524
Villalta M, Estévez A, Bransden MP, Bell JG (2005) The effect of graded concentrations of dietary DHA on growth, survival and tissue fatty acid profile of Senegal sole (Solea senegalensis) larvae during the Artemia feeding period. Aquaculture 249:353–365
Wan J, Xiao Z, Chao S, Xiong S, Gan X, Qiu X, Xu C, Ma Y, Tu X (2014) Pioglitazone modulates the proliferation and apoptosis of vascular smooth muscle cells via peroxisome proliferators-activated receptor-gamma. Diabetology & Metabolic Syndrome 6:101
Wang AS, Xu CW, Xie HY, Yao AJ, Shen YZ, Wan JJ, Zhang HQ, Fu JF, Chen ZM, Zou ZQ (2016) DHA induces mitochondria-mediated 3T3-L1 adipocyte apoptosis by down-regulation of Akt and ERK. J Funct Foods 21:517–524
Wang X, Huang M, Wang Y (2012) The effect of insulin, TNFα and DHA on the proliferation, differentiation and lipolysis of preadipocytes isolated from large yellow croaker (Pseudosciaena crocea R.) PloS One 7:e48069
Watanabe T (2007) Importance of docosahexaenoic acid in marine larval fish. J World Aquacult Soc 24:152–161
Yang C, Zhang GP, Chen YN, Meng FL, Liu SS, Gong SP (2016) Effects of docosahexaenoic acid on cell apoptosis, invasion and migration of cervical cancer cells in vitro. Journal of Southern Medical University 36:848–856
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This study was supported financially by the National Natural Science Foundation of China (31372538). We thank Ankang Fisheries Experimental and Demonstration Station (AFEDS) of the Northwest Agriculture and Forestry University for the experimental fish.
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Jin, A., Lei, Cx., Tian, Jj. et al. Dietary docosahexaenoic acid decreased lipid accumulation via inducing adipocytes apoptosis of grass carp, Ctenopharygodon idella . Fish Physiol Biochem 44, 197–207 (2018). https://doi.org/10.1007/s10695-017-0424-6
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DOI: https://doi.org/10.1007/s10695-017-0424-6