اثر منبع و مدت زمان تغذیه اسید چرب امگا-3 و امگا-9 محافظت شده بر بیان برخی از ژن های دخیل در سوخت و ساز چربی در بره های پرواری

نوع مقاله : مقاله پژوهشی

نویسندگان

1 استادیار پژوهشی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان مرکزی، اراک، ایران

2 دانشیار، گروه علوم دامی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران

3 استادیار، گروه علوم دامی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران

چکیده

هدف از مطالعه حاضر بررسی تأثیر استفاده از نمک­های کلسیمی روغن ماهی، زیتون و چربی اشباع بر بیان برخی از ژن­های مرتبط با سوخت و ساز چربی در بره­های پرواری آمیخته لری بختیاری×رومانف بود. تعداد 49 رأس بره نر چهار تا پنج ماهه با میانگین وزن اولیه 88/0±97/29 کیلوگرم، در قالب یک طرح کاملاً تصادفی به هفت گروه هفت رأسی تقسیم شدند. تیمارهای مورد بررسی شامل: جیره شاهد (بدون چربی) و جیره پایه به همراه نمک­های کلسیمی روغن ماهی، روغن زیتون و چربی اشباع (به میزان دو درصد ماده خشک جیره) برای دوره­های زمانی 45 و 90 روزه مصرف بودند. در انتهای آزمایش، تعداد 28 رأس بره ذبح شده و یک نمونه از کبد بره­ها گرفته شد تا به وسیله آن، مطالعات نوتری­ژنومیک انجام شود. نتایج نشان داد که استفاده از نمک­های کلسیمی روغن ماهی، روغن زیتون و چربی اشباع، تأثیر معنی­داری بر بیان ژن­های FADS1 و FADS2 در بافت کبد نسبت به گروه شاهد نداشت. با این حال، استفاده از نمک­های کلسیمی روغن ماهی و روغن زیتون در دوره­های زمانی 90 و 45 روزه مصرف باعث افزایش معنی­دار بیان mRNA کبدی ژن­های CPT1 (001/0=P) و ACOX1 (002/0=P) در بافت کبد نسبت به تیمارهای چربی اشباع و شاهد شد. نتایج حاصل از این مطالعه نشان داد که مکمل­ سازی جیره با نمک­های کلسیمی اسیدهای چرب غیراشباع ماهی و زیتون صرف­نظر از دوره زمانی مصرف، باعث افزایش بیان ژن­های درگیر در لیپولیز چربی­های بافت کبد بره ­ها شد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Effect of source and duration of feeding omega-3 and omega-9 protected fatty acids on the expression of some genes involved in fat metabolism in fattening lambs

نویسندگان [English]

  • A. Mirshamsollahi 1
  • M. ganjkhanlou 2
  • F. Fatehi 3
1 Assistant Professor, Agriculture and Natural Resources Research Center of Markazi province, Arak, Iran
2 Associate Professor, Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
3 Assistant Professor, Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
چکیده [English]

Introduction: Based on current knowledge, it has been proven that many edible ingredients in addition to meeting the needs of creatures, in terms of energy and protein, contain compounds that affect cellular actions and intracellular signaling pathways and can temporarily or permanently change the function of the cell. The use of molecular biology tools and genetic research defines the mechanisms through which gene expression is affected by food, and conversely, these genes also affect the absorption of food, metabolism, and excretion. From the point of view of nutrigenomics, dietary nutrients are signals that are received by sensitive cell systems and can affect the expression of genes and proteins and the production of metabolites. Nutritional manipulations and strategies are key tools to influence ruminant production. Fatty acids act on the nucleus by binding to and regulating the activity of specific nuclear receptors or transcription factors, thus playing a central role in regulating the expression of genes involved in fatty acid uptake by muscle cells. Interactions between diet nutrients and the expression of genes involved in lipid metabolism have many possibilities regarding the deposition of fatty acids in the tissue. The study of gene expression has enabled clarification of the mode of fatty acid metabolism in muscle and the accumulation of intramuscular fat or marbling and the role of the genes that promote fatty acid oxidation and mitochondrial respiration in the liver muscle and adipose tissue. Polyunsaturated fatty acids with multiple double bonds (PUFA) and monounsaturated fatty acids with one double bond (MUFA) have received much attention in the last decade, and their health benefits are increasingly evident. Therefore, this study investigated the effect of using calcium salts of fish oil, olive oil, and saturated fat on the expression of some genes related to fat metabolism in Lori Bakhtiari×Romanov fattening lambs.
Materials and methods: This study was carried out at the educational research station of the Department of Animal Science of the College of Agriculture and Natural Resources, University of Tehran. 49 male lambs aged four to five months with an average initial weight of 29.97 ± 0.88 kg were divided into seven groups of seven lambs in a completely randomized design. The experiment consisted of seven treatments with a basic diet as follows: 1) Basic diet without fat powder (control), 2 and 3) Basic diet with calcium salt of fish oil (rich in omega-3 fatty acids) in the amount of 2% dry matter of the diet for 90 and 45 days, respectively, 4 and 5) Basic diet with calcium salt of olive oil (rich in omega-9 fatty acids) at the rate of 2% dry matter of the diet for 90 and 45 days, respectively, 6 and 7) Basic diet with saturated fat powder in the amount of 2% dry matter of the diet for 90 and 45 days, respectively. Rations were adjusted based on the NRC Sheep and Goat and using the fifth version of CNCPS (The Cornell Net Carbohydrate and Protein System) software so that they are the same in terms of energy and protein. Twenty-eight lambs were slaughtered and a liver sample was taken for nutrigenomics studies. Total RNA was extracted from liver tissue samples using the RNA extraction kit produced by Dena Bio Asia Company (Mashhad) and according to the protocol provided with the kit. The expression of four main genes involved in lipid metabolism in liver tissue was investigated to provide comprehensive information on the effects of omega-3, omega-9 fatty acids, and saturated fat at the molecular level.
Results and discussion: The results showed that feeding with calcium salts of fish oil, olive oil, and saturated fat, both in the whole 90 days or the last 45 days of the fattening period, had no significant effect on the expression of FADS1 and FADS2 genes in liver tissue compared to the control group. However, the use of calcium salts of fish and olive oils in periods of 90 and 45 days significantly increased the hepatic mRNA expression of the CPT1 gene (P=0.001) and ACOX1 gene mRNA expression in liver tissue compared to control and saturated fat treatments (P=0.002). The CPT1 enzyme is responsible for a mitochondrial transport system and plays a key role in controlling the oxidation of long-chain fatty acids and stimulating mitochondrial β-oxidation.
Conclusions: The results of this study showed that diet supplementation with calcium salts of fish and olive unsaturated fatty acids, regardless of the period of consumption, increased the expression of genes involved in the lipolysis of liver tissue fats of lambs.

کلیدواژه‌ها [English]

  • Unsaturated fatty acid
  • Fattening lamb
  • Gene expression
  • Metabolism
Abrahams, V. (2017). Novel mechanisms of placental inflammation in obstetric antiphospholipid syndrome. Placenta, 57, 243.
Arana, A., Mendizabal, J. A., Alz´on, M., Eguinoa, P., Beriain, M. J., & Purroy, A. (2006). Effect of feeding lambs oleic acid calcium soaps on growth, adipose tissue development and composition. Small Ruminant Research, 63, 75-83. doi:10.1016/j.smallrumres.2005.02.006
Belal, S. A., Subramaniyan Sivakumar, A., Kang, D. R., Cho, S., Choe, H. S., & Shim, K. S. (2018). Modulatory effect of linoleic and oleic acid on cell proliferation and lipid metabolism gene expressions in primary bovine satellite cells. Animal Cells and Systems, 22(5), 324–333. doi: 10.1080/19768354.2018.1517824
Bonnefont, J. P., Djouadi, F., Prip-Buus, C., Gobin, S., Munnich, A., & Mol, B. J. (2004). Carnitine palmitoyltransferases 1 and 2: biochemical, molecular and medical aspects. Molecular Aspects of Medicine, 25, 495–520. doi: 10.1016/j.mam.2004.06.004
Cherfaoui, M., Durand, D., Bonnet, M., Bernard, L., Bauchart, D., Ortigues-Marty, I., & Gruffat, D. (2013). A grass-based diet favours muscle n-3 long-chain PUFA deposition without modifying gene expression of proteins involved in their synthesis or uptake in Charolais steers. Animal, 7, 1833-1840. doi: 10.1017/S1751731113001432
Chomczynski, P., & Sacchi, N. (2006). The single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on. Nature Protocols1, 581–585. doi: 10.1038/nprot.2006.83  
Clarke, S. (2000). Polyunsaturated fatty acid regulation of gene transcription: a mechanism to improve energy balance and insulin resistance. British Journal of Nutrition, 83, S59-S66. doi: 10.1017/s0007114500000969
Clarke, S. D. (2001). Nonalcoholic steatosis and steatohepatitis. I. Molecular mechanism for polyunsaturated fatty acid regulation of gene transcription. American Journal of Physiology, 281, 865-869. doi: 10.1152/ajpgi.2001.281.4. G865
Coleman, D. N., Carranza Martin, A. C., Jin, Y., Lee, K., & Relling, A. E. (2019). Prepartum fatty acid supplementation in sheep. IV. Effect of calcium salts with eicosapentaenoic acid and docosahexaenoic acid in the maternal and finishing diet on lamb liver and adipose tissue during the lamb finishing period. Journal of Animal Science, 97(7), 3071-3088. doi: 10.1093/jas/skz154
Costa Alvarenga, T. I. R., Chen, Y., Furusho-Garcia, I. F., Olalquiaga Perez, J. R., & Hopkins, D. L. (2015). Manipulation of Omega-3 PUFAs in Lamb: Phenotypic and Genotypic Views. Comprehensive Reviews in Food Science and Food Safety, 14, 189-204. doi: 10.1111/1541-4337.12131
Costa Alvarenga, T. I. R., Chen, Y., Lewandowski, P., Ponnampalam, E. N., Sediq, S., Clayton, E. H., van de Ven, R. J., Olalquiaga Perez, J. R., & Hopkins, D. L. (2016). The expression of genes encoding enzymes regulating fat metabolism is affected by maternal nutrition when lambs are fed algae high in omega-3. Livestock Science, 187, 53-60. doi: 10.1016/j.livsci.2016.02.013
Dervishi, E., Serrano, C., Joy, M., Serrano, M., Rodellar, C., & Calvo, J. H. (2010). Effect of the feeding system on the fatty acid composition, expression of the Δ9-desaturase, Peroxisome Proliferator-Activated Receptor Alpha, Gamma, and Sterol Regulatory Element Binding Protein 1 genes in the semitendinous muscle of light lambs of the Rasa Aragonesa breed. BMC Veterinary Research, 6, 40-51. doi:10.1186/1746-6148-6-40
Dervishi, E., Serrano, C., Joy, M., Serrano, M., Rodellar, C., & Calvo, J. H. (2011). The effect of the feeding system in the expression of genes related with fat metabolism in semitendinous muscle in sheep. Meat Science, 89, 91-97. doi: 10.1016/j.meatsci.2011.04.003
Fekri, A., & Ganj Khanlu, M. (2021). Effects of supplemental omega-3 protected sources of fish oil and flaxseed oil on ruminant performance. Domestic Journal, 20 (18), 48-53. doi: 10.22059/domesticsj.2021.312293.1053 [In Persian]
Gregory, M. K., Gibson, R. A., Cook-Johnson, R. J., Cleland, L. G., & James, M. J. (2011). Elongase reactions as control points in long-chain polyunsaturated fatty acid synthesis. PLoS One, 6(12), e29662. doi: 10.1371/journal.pone.0029662
Hashemzadeh, F., Rafeie, F., Hadipour, A., & Rezadoust, M. H. (2023). Effect of adding a phytogenic-rich herbal mixture to diet on the expression pattern of some insulin hormone metabolism-related candidate genes of heat-stressed fattening Afshari-Shal lambs. Animal Production Research, 12(1), 25-37. doi: 10.22124/AR.2023.21822.1690 [In Persian]
Haug, A., Nyquist, N. F., Thomassen, M., Høstmark, A. T., & Østbye, T. K. (2014). N-3 fatty acid intake altered fat content and fatty acid distribution in chicken breast muscle, but did not influence mRNA expression of lipid-related enzymes. Lipids in Health and Disease, 13, 92. doi: 10.1186/1476-511X-13-92   
Iommelli, P., Infascelli, F., Musco, N., Grossi, M., Ferrara, M., Sarubbi, F., D’Aniello, B., Lombardi, P., & Tudisco, R. (2021). Stearoyl-CoA desaturase activity and gene expression in the adipose tissue of buffalo bulls was unaffected by diets with different fat content and fatty acid profile. Agriculture, 11, 1209-1221. doi: 10.3390/agriculture11121209
Jager, N., Hudson, N. J., Reverter, A., Barnard, R., Café, L. M., Greenwood, P. L, & Dalrymple, B. P. (2013). Gene expression phenotypes for lipid metabolism and intramuscular fat in skeletal muscle of cattle. Journal of Animal Science, 91, 1112-1128. doi: 10.2527/jas.2012-5409
Kopecky, J., Rossmeisl, M., Flachs, P., Kuda, O., Brauner, P., Jilkova, Z., Stankova, B., Tvrzicka, E., & Bryhn, M. (2009). Symposium on ‘Frontiers in adipose tissue biology’ n-3 PUFA: bioavailability and modulation of adipose tissue function. Proceedings of the Nutrition Society, 68, 361-369. doi: 10.1017/S0029665109990231
Ladeira, M. M., Schoonmaker, J. P., Swanson, K. C., Duckett, S. K., Gionbelli, M. P., Rodrigues, L. M., & Teixeira, P. D. (2018). Review: Nutrigenomics of marbling and fatty acid profile in ruminant meat. Animal, 12(2), 282-294. doi:10.1017/S1751731118001933
Lim, J. H., Gerhart-Hines, Z., Dominy, J. E., Lee, Y., Kim, S., Tabata, M., Xiang, Y. K., & Puigserver, P. (2013). Oleic acid stimulates complete oxidation of fatty acids through protein kinase a-dependent activation of SIRT1-PGC1α complex. The Jourrnal of Biological Chemistry, 288(10), 7117-7126. doi: 10.1074/jbc.M112.415729
Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 25(4), 402-408. doi: 10.1006/meth.2001.1262
Matsumoto, H., Nogi, T., Tabuchi, I., Oyama, K., Mannen, H., & Sasazaki, S. (2014). The SNPs in the promoter regions of the bovine FADS2 and FABP4 genes are associated with beef quality traits. Livestock Science, 163, 34-40. doi: 10.1016/j.livsci.2014.02.016
Mirshamsollahi, A. Ganjkhanlou, M., Fatehi, F., Zali, A., & Sadeghi, M. (2022). Effect of source and duration of protected fatty acids feeding, on production performance, carcass characteristics and blood metabolites in fattening lambs. Research on Animal Production, 13(36), 47-56. doi:10.52547/rap.13.36.47 [In Persian]
NRC. (2007). Nutrient Requirments of Small ruminant (7th ed.). National Academy Press, Washington, DC.
Ponnampalam, E. N., Lewandowski, P. A., Fahri, T., Fahri, V. F., Burnett, F. R., Dunshea, T. Plozza, J., & Jacobs, L. (2015). Forms of n‑3 (ALA, C18:3n‑3 or DHA, C22:6n‑3) fatty acids affect carcass yield, blood lipids, muscle n‑3 fatty acids and liver gene expression in lambs. Lipids, 50(11), 1133-1143. doi: 10.1007/s11745-015-4070-4
Radonic, A., Thulke, S., Mackay, I. M., Landt, O., Siegert, W., & Nitsche, A. (2004). Guideline to reference gene selection for quantitative real-time PCR. Biochemical and Biophysical Research Communications, 313, 856-862. doi: 10.1016/j.bbrc.2003.11.177
Smith, S. B., Lunt, D. K., Chung, K. Y., Choi, C. B., Tume, R. K., & Zembayashi, M. (2006). Adiposity, fatty acid composition, and delta-9 desaturase activity during growth in beef cattle. Animal Science Journal, 77, 478-486. doi: 10.1111/j.1740-0929.2006.00375x
Toral, P. G., Bernard, L., Delavaud, C., Gruffat, D., Leroux, C., & Chilliard, Y. (2013). Effects of fish oil and additional starch on tissue fatty acid profile and lipogenic gene mRNA abundance in lactating goats fed a diet containing sunflower-seed oil. Animal, 7, 948–956. doi: 10.1017/S1751731113000049
Untergasser, A., Nijveen, H., Rao, X., Bisseling, T., Rene ́Geurts, R., & Leunissen, A. M. (2007). Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Research, 35, 71-74. doi: 10.1093/nar/gkm306
Vahmani, P., Glover, K. E., & Fredeen, A. H. (2014). Effects of pasture versus confinement and marine oil supplementation on the expression of genes involved in lipid metabolism in mammary, liver, and adipose tissues of lactating dairy cows. Journal of Dairy Science, 97, 4174-4183. doi: 10.3168/jds.2013-7290
Vassiliou, E. K., Gonzalez, A., Garcia, C., Tadros, J. H., Chakraborty, G., & Toney, J. H. (2009). Lipids in Health and Disease, 8, 25. doi: 10.1186/1476-511X-8-25
Ye, J., Coulouris, G., Zaretskaya, I., Cutcutache, I., Rozen, S., & Madden, T. (2012). Primer-BLAST: A tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics, 13, 134-145. doi:10.1186/1471-2105-13-13