تاثیر افزودن مکمل آلی روی-متیونین و مکمل نمک کلسیمی روغن کتان بر بیان ژ‌‌ن‌IGF-1 در بافت کبد بره‌های نر پرواری

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشیار. گروه علوم دامی;دانشکده علوم دامی و صنایع غذایی -دانشگاه علوم کشاورزی و منابع طبیعی خوزستان-ملاثانی-ایران

2 دانشجوی دکتری. گروه علوم دامی;دانشکده علوم دامی و صنایع غذایی -دانشگاه علوم کشاورزی و منابع طبیعی خوزستان-ملاثانی-ایران

3 دانشجوی دکتری ژنتیک و اصلاح نژاد دام، گروه علوم دامی، دانشکده علوم دامی و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی خوزستان، ملاثانی، ایران.

چکیده

هدف: عنصر روی یکی از محدودکننده‌ترین ریزعناصر معدنی است که برای رشد بدن، ساختمان و فعالیت هورمونی و آنزیمی، سوخت ‌و ساز مواد مغذی، تقسیم سلولی و سیستم ایمنی مورد نیاز است. مشخص شده که کمبود روی با کاهش مصرف غذا، کاهش رشد و کاهش تولید IGF-1 در کبد همراه است. عنصر روی یکی از عوامل مهم در تنظیم بیان ژن خانواده IGF در بسیاری از بافت­ها است. همچنین عنصر روی بر غیر اشباع‌سازی اسید لینولئیک موثر است. عنصر روی می‌تواند از پراکسیداسیون لیپیدها جلوگیری کند. به طور کلی ارتباط قابل توجهی بین متابولیسم چربی و عنصر روی وجود دارد. از طرف دیگر روغن کتان حاوی اسیدچرب ضروری آلفالینولینک اسید (امگا3) است که به نظر می‌رسد با عملکردهای IGF-1 در بدن مرتبط باشد. لذا در این پژوهش تاثیر افزودن مکمل آلی روی-متیونین به جیره‌های با و بدون مکمل نمک کلسیمی روغن کتان بر بیان ژن‌ IGF-1 در بافت کبد بره­های نر پرواری مورد بررسی قرار گرفت.
مواد و روش­ها: در این پژوهش از 44 راس بره نر عربی با آزمایش فاکتوریل 2*2 در قالب طرح کاملا تصادفی با چهار تیمار و 11 تکرار استفاده شد. چهار جیره‌ی آزمایشی عبارت بودند 1) جیره پایه بدون مکمل نمک کلسیمی روغن کتان و بدون مکمل روی- متیونین (کنترل)، 2) جیره پایه بدون مکمل کلسیمی روغن کتان حاوی 08/0 درصد مکمل روی- متیونین (معادل 100 میلی‌گرم روی در کیلوگرم ماده خشک، 3) جیره پایه حاوی 3 درصد مکمل نمک کلسیمی روغن کتان بدون مکمل روی-متیونین ، 4) جیره پایه حاوی 3 درصد مکمل نمک کلسیمی روغن کتان بعلاوه 08/0 درصد مکمل روی-متیونین. پس از پایان دوره پروار 3 راس گوسفند از هر تیمار کشتار گردید و بافت کبد همراه با ازت مایع به آزمایشگاه منتقل شد. پس از استخراج RNA و اندازه‌گیری کیفیت آن، سنتز cDNA انجام شد. در نهایت میزان بیان ژن IGF-1 با استفاده از روش Real time PCR مورد ارزیابی قرار گرفت.
نتایج: مشاهده تک باند در محدوده‌ی 240 جفت نوکلئوتید برای ژن IGF-1 و در محدوده‌ی 144 جفت نوکلئوتید برای ژن GAPDH بر روی الکتروفورز ژل، بیانگر صحت انجام آزمایش و تکثیر قطعه‌ی مورد نظر بوسیله واکنش زنجیره‌ای پلیمراز بود. حضور تنها یک قله در منحنی‌های ذوب ژن IGF-1 و GAPDH در واکنش Real time PCR تولید یک محصول اختصاصی در این واکنش را تایید کرد. نتایج حاصل از این پژوهش نشان داد، افزودن مکمل روی منجر به افزایش معنی‌دار بیان ژن IGF-1 شده (01/0 >P)، به طوری که از 1 به 95/4 رسیده است. همچنین اثر افزودن مکمل چربی نمک کلسیمی روغن کتان بر بیان ژن IGF-1 معنی­دار یوده (01/0 >P) به طوری که از 1 به 52/3 رسیده است. اثرات متقابل مکمل روی- متیونین و مکمل چربی نمک کلسیمی روغن کتان معنی‌دار نشد. افزودن همزمان مکمل روی- متیونین و مکمل چربی نمک کلسیمی روغن کتان به جیره بره‌های پرواری سبب افزایش بیان ژن IGF-1 در کبد می‌گردد.
نتیجه­ گیری: افزودن مکمل آلی روی-متیونین به جیره‌های حاوی مکمل چربی نمک کلسیمی روغن کتان سبب افزایش بیان ژن‌ IGF-1 می‌گردد. افزایش بیان ژن‌ IGF-1 در کبد احتمالا منجر به افزایش رشد و در نهایت تولید دام خواهد شد.

کلیدواژه‌ها


عنوان مقاله [English]

Effect of zinc-methionine organic and Ca-salt of flaxseed oil supplement on IGF-1 gene expression in liver tissue of fattening male lambs

نویسندگان [English]

  • Mahmood Nazari 1
  • Zeinab Alipoor 2
  • Sepideh Rostami 3
  • Ghazal Mohamadi Ahvazi 3
1 Associate Professor, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Mollasani, Iran.
2 Ph. D student, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Mollasani, Iran.
3 Ph. D student, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Mollasani, Iran.
چکیده [English]

Objective
Zinc is one of the most limiting trace mineral elements required for body growth, structure, hormonal and enzyme activity, nutrient metabolism, cell division, and immune system function. Zinc deficiency has been associated with reduced food intake, stunted growth, and decreased production of IGF-1 in the liver. Zinc plays a key role in the regulation of IGF family gene expression in various tissues and is also effective in desaturating linoleic acid, thereby preventing lipid peroxidation. There is a significant relationship between fat metabolism and zinc. On the other hand, flaxseed oil contains the essential fatty acid alpha-linolenic acid (omega-3), which appears to be related to IGF-1 function in the body. Therefore, this research investigated the effect of adding a zinc-methionine organic supplement to diets with and without the calcium salt of flaxseed oil on IGF-1 gene expression in the liver tissue of fattening lambs.
Materials and Methods
In this research, 44 Arab male lambs were used in a 2 x 2 factorial experiment within a completely randomized design, with four treatments and 11 replications. The four experimental diets were: 1) basic diet without Ca-salt of flaxseed oil supplement and without zinc-methionine supplement (CON), 2) basic diet without Ca-salt of flaxseed oil supplement containing 0.08% zinc-methionine supplement, 3) basic diet containing 3% Ca-salt of flaxseed oil supplement without zinc-methionine supplement, and 4) basic diet containing 3% Ca-salt of flaxseed oil supplement plus 0.08% zinc-methionine supplement. After the fattening period, three lambs from each treatment were slaughtered, and muscle tissue was transferred to the laboratory with liquid nitrogen. After RNA extraction and quality assessment, cDNA synthesis was performed. The expression of lipogenic genes was evaluated using the Real-time PCR method.
Results
The presence of a single band in the range of 240 base pairs for the IGF-1 gene and 144 base pairs for the GAPDH gene on gel electrophoresis confirmed the accuracy of the test and the correct amplification of the desired fragments by polymerase chain reaction. The presence of a single peak in the melting curves of the IGF-1 and GAPDH genes in the real-time PCR reaction further confirmed the specificity of the produced products. The results showed that the addition of zinc supplements significantly increased the expression of the IGF-1 gene (P < 0.01), from 1 to 4.95. Additionally, the calcium salt of flaxseed oil supplement significantly increased IGF-1 gene expression (P < 0.01), raising it from 1 to 3.52. The interaction effects of the zinc-methionine supplement and calcium salt flax oil supplement were not significant. Simultaneous addition of the zinc-methionine supplement and the calcium salt flax oil supplement to the diet of fattening lambs increased IGF-1 gene expression in the liver.
Conclusions
The addition of a zinc-methionine organic supplement to diets containing a calcium salt flaxseed oil supplement increases IGF-1 gene expression. Increasing IGF-1 gene expression will likely enhance growth and overall production.

کلیدواژه‌ها [English]

  • flaxseed oil
  • gene expression
  • IGF-1
  • zinc
سیدآبادی حمیدرضا، عسگری جعفرآبادی قباد، محمودی امیررضا، جواهری بارفروش هدی (1400) تاثیر استفاده از دانه کتان بعنوان منبع امگا 3 برمیزان تولید شیر و بیان ژن های درگیر در رشد و نمو بافت پستان در بزهای سانن. نشریه دامپزشکی ایران 133، 44-32.
ربیعه مولود، روشنفکر هدایت‌الله، نظری محمود، قربانی محمدرضا (1400) بررسی بیان ژن آنزیم‌های آنتی‌اکسیدانی در جوجه‌های گوشتی تغذیه شده با ویتامین E، عصاره‌ی گیاه خرفه و پسته‌ی وحشی تحت استرس گرمایی. نشریه دامپزشکی ایران 17(2)، 60-51.
نظری محمود، سالاری سمیه، قربانی محمدرضا (1396) اثر مکمل عنصر روی و جایگزینی بتائین با بخشی از متیونین بر بیان ژن بتایین هموسیستئین متیل ترانسفراز و ژنهای لیپوژنیک کبدی مرغ تخمگذار تحت شرایط تنش گرمایی. مجله بیوتکنولوژی کشاورزی 9(1)، 110-95.
نظری محمود، سالاری سمیه، قربانی محمدرضا (1399) اثر مکمل روی و جایگزینی بتائین جیره با متیونین بر عملکرد و فراسنجه‌های خونی مرغان تخمگذار در شرایط استرس گرمایی. تحقیقات دامپزشکی و فرآورده‌های بیولوژیک 33(1)، 70-61.
 
Amiri Roudbar M, Mohammadabadi MR, Ayatollahi Mehrgardi A, et al. (2020) Integration of single nucleotide variants and whole-genome DNA methylation profiles for classification of rheumatoid arthritis cases from controls. Heredity 124 (5), 658-674.
Andrieu S (2008) Is there a role for organic trace element supplements in transition cow health? Vet J 176, 77–83.
Barazandeh A, Mohammadabadi MR, Ghaderi-Zefrehei M, Nezamabadi-Pour H (2016) Genome-wide analysis of CpG islands in some livestock genomes and their relationship with genomic features. Czech J Anim Sci 61(11), 487–495.
Bordbar F, Mohammadabadi M, Jensen J, et al. (2022) Identification of candidate genes regulating carcass depth and hind leg circumference in Simmental beef cattle using Illumina Bovine Bead chip and next-generation sequencing. Animals 12 (9), e1103.
Chalupa WB, Vecchiarelli B, Sklan D, Kronfeld, DS (1985) Response of rumen microorganisms and lactating cows to calcium salts of long chain fatty acids. J Dairy Sci 68 (Suppl.1), e110.
Devine A, Rosen C, Mohan S, et al. (1998) Effects of zinc and other nutritional factors on insulin-like growth factor I and insulin-like growth factor binding proteins in postmenopausal women. Am J Clinic Nutr 68(1), 200-206.
Dirandeh E, Towhidi A, Ansari Z, et al. (2016) Effects of dietary supplementation with different polyunsaturated fatty acids on expression of genes related to somatotropic axis function in the liver, selected blood indicators, milk yield and milk fatty acids profile in dairy cows. Annals of Anim Sci 16(4), 1045-1058.‏
Droke EA, Spears JW, et al. (1993) in vitro and in vivo immunological measurements in growing lambs fed diets deficient, marginal or adequate in zinc. J Nutri Immuno 2, 71-90.
Fair S, Doyle DN, Diskin MG, et al. (2014) Theeffect of dietary n-3 polyunsaturated fatty acids supplementation oframs on semen quality and subsequent quality of liquid storedsemen. Theriogenology 81(2), 210–219.
Fenwick MA, Fitzpatrick R, Kenny DA, et al. (2008) Interrelationships between negative energy balance (NEB) and IGF regulation in liver of lactating dairy cows. Domest Anim Endocrin 34(1), 31– 44.
Gerard S, David A, Sinead M (2011) Effect of dietary n-3 polyunsaturated fatty acid supplementation on bovine uterine endometrial and hepatic gene expression of the insulin-like growth factor system. Therogenology 75, 500-512.
Greene LW, Lunt DK, Byers FM, et al. (1988) Performance and carcass quality of steers supplemented with zinc oxide or zinc methionine. J Anim Sci 66(7), 1818-1823.‏
Guernec A, Chevalier B, Duclos MJ (2004) Nutrient supply enhances both IGF-I and MSTN mRNA levels in chicken skeletal muscle. Domest Anim Endocrin 26, 143-54.
Haase H, Maret W (2003) Intracellular zinc fluctuations modulate protein tyrosine phosphatase activity in insulin/insulin-like growth factor-1 signaling. Exp Cell Res 291, 289–98.
Heck A, Metayer S, Onagbesan OM, Williams J (2003) mRNA expression of components of the IGF system and of GH and insulin receptors in ovaries of broiler breeder hens fed ad libitum or restricted from 4 to 16 weeks of age. Domest Anim Endocrin 25, 287-94.
Jafari Ahmadabadi SAA, Askari-Hemmat H, Mohammadabadi M, et al. (2023) The effect of Cannabis seed on DLK1 gene expression in heart tissue of Kermani lambs. Agric Biotech J 15 (1), 217-234.
Jafarpour N, Khorvash M, Rahmani HR, et al. (2015) Dose–responses of zinc–methionine supplements on growth, blood metabolites and gastrointestinal development in sheep. J Anim Physio and Anim Nutr 99, 668–675.
Kakilashvili B, Zurabashvili DZ, Turabelidze DG, et al. (2014) The fatty acid composition of ordinary flax seed oil (Linum usitatissimum L.) cultivated in Georgia and its byological activity. Georgian medical news 227, 86-88.
Khabiri A, Toroghi R, Mohammadabadi M, Tabatabaeizadeh SE (2023) Introduction of a Newcastle disease virus challenge strain (sub-genotype VII. 1.1) isolated in Iran. Vet Res Forum 14 (4), e221.
Kronberg SL, Scholljegerdes EJ, Lepper AN, Berg EP (2011) The effect of flaxseed supplementation on growth, carcass characteristics, fatty acid profile, retail shelf life and sensory characteristics of beef from steers finished on grasslands of the northern Great Plains. J Anim Sci 89(9), 2892-2903.
LeRoith D, Yakar S (2007) Mechanisms of disease: metabolic effects of growth hormone and insulin-like growth factor 1. Nat Clin Pract Endocrinol Metab 3, 302-10.
Li X, Yin J, Li D, et al. (2006) Oxide Increases IGF-I and IGF-I Receptor Gene Expression in the Small Intestine of Weanling Piglets. J Nutr 136, 1786–1791.
MacDonald R.S (2000) The Role of Zinc in Growth and Cell Proliferation. J Nutr 130, 1500S-1508S.
 Matar M, Al-Shaar L, Maalouf J, et al. (2016) The Relationship Between Calciotropic Hormones, IGF-1, and Bone Mass Across Pubertal Stages. J clin endo and meta 101(12), 4860-4870.
Meyer Z, Höflich C, Wirthgen E, et al. (2017) Analysis of the IGF-system in milk from farm animals–Occurrence, regulation, and biomarker potential. Growth Hormone IGF Res 35, 1-7‏.
Miltko R, Majewska M P, Bełżecki G. et al. (2019) Growth performance, carcass and meat quality of lambs supplemented different vegetable oils. Asian-Australasian J Anim Sci 32(6), 767.
Mohamadinejad F, Mohammadabadi M, Roudbari Z, et al. (2024) Analysis of liver transcriptome data to identify the genes affecting lipid metabolism during the embryonic and hatching periods in ROSS breeder broilers. J Livest Sci Technol 12(2), 61-67.
Mohammadabadi M, Babenko O, Borshch OO, et al. (2024) Measurement of the relative expression pattern of the UCP2 gene in different tissues of the Raini Cashmere goat. Agric Biotech J 16 (3), 317-332.
Mohammadabadi M, Golkar A, Askari Hesni M (2023) The effect of fennel (Foeniculum vulgare) on insulin-like growth factor 1 gene expression in the rumen tissue of Kermani sheep. Agric Biotech J 15, 239-256.
Mohammadabadi MR (2016) Inter-Simple Sequence Repeat Loci associations with predicted breeding values of body weight in Kermani sheep. Genet 3rd Millennium 14, 4383-4390.
Mohammadinejad F, Mohammadabadi M, Roudbari Z, Sadkowski T (2022) Identification of Key Genes and Biological Pathways Associated with Skeletal Muscle Maturation and Hypertrophy in Bos taurus, Ovis aries, and Sus scrofa. Animals 12, e3471.
Mohammed AA, Al-Shaheen T, Al-Saiady M, El-Waziry A (2024) Effect of dietary source of omega-3 fatty acids on milk production, fatty acid profiles and IGF-1 of lactating dairy cows in arid subtropics. Pakistan J Zoo 56(5), ‏2319-2326.
National Research Council, NRC (2007) Nutrient Requirements of Small Ruminants: Sheep, Goats, Cervids, and New World Camelids. The National Academies Press, Washington DC, USA.
Nazari M, Salari S, Ghorbani MR (2017) Effects of zinc supplementation and betaine substitution to methionine on hepatic betaine - homocysteine methyltransferase and lipogenic genes expression in laying hens under heat stress. Agric Biotech J 9(1), 95-110 (In Persian)
Nazari M, Sallari S, Ghorbani MR (2020) Effect of Zinc supplementation and Betaine substitution to methionine on performance and blood parameters of laying hens under heat stress. Vet Res & Bio Prod 33(1), 61-70 (In Persian)
Ninh NX, Thissen JP, Maiter D, et al. (1995) Reduced liver insulin-like growth factor-I gene expression in young zinc-deprived rats is associated with a decrease in liver growth hormone (GH) receptors and serum GH-binding protein. J Endocrin 144, 449–56.
Pfaffl M, Horgan G, Dempfle L (2002) Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nuc Acids Res 30, 1-10.
Ponnampalam EN, Priyashantha H, Vidanarachchi JK, et al. (2024) Effects of Nutritional Factors on Fat Content, Fatty Acid Composition, and Sensorial Properties of Meat and Milk from Domesticated Ruminants: An Overview. Animals (Basel) 14(6), 840.
Rabieh MH, Rooshanfekr M, Nazari M, Ghorbani MR (2020) Gene expression of antioxidant enzymes fed wild pistachio (Pistachio atlantica), purslane (portulaca oleracea) extract and vitamin E under in broiler chickens under heat stress condition. Iranian Vet J 17(2), 51-60 (In Persian).
Radcliff RP, McCormack BL, Crooker BA, Lucy MC (2003( Growth hormone (GH) binding and expression of GH receptor 1AmRNA in hepatic tissue of periparturient dairy cows. J Dairy Sci 86, 3933– 40.
Rashidi AA, Ivari YG, Khatibjoo A, Vakili R (2010) Effects of dietary fat, vitamin E and zinc on immune response and blood parameters of broiler reared under heat stress. Res J Poultry Sci 3(2), 32-38.
Reyna XF, Montoya HM, Castrellon VV, et al. (2010) Polymorphisms in the IGF1 gene and their effect on growth traits in Mexican beef cattle. Gene and Mol Res 9(2), 875-83.
Rhoads ML, Meyer JP, Lamberson WR, et al. (2008) Uterine and hepatic gene expression in relation to days postpartum estrus and pregnancy in postpartum dairy cows. J Dairy Sci 91, 140 –50.
Safaei SMH, Dadpasand M, Mohammadabadi M, et al. (2022) An Origanum majorana Leaf Diet Influences Myogenin Gene Expression, Performance, and Carcass Characteristics in Lambs. Animals 13, e14
Safaei SMH, Mohammadabadi M, Moradi B, et al. (2023) Role of Fennel (Foeniculum vulgare) Seed Powder in Increasing Testosterone and IGF1 Gene Expression in the Testis of Lamb. Gene Expr 4, 1-20.
Salabi F, Boujarpoor M, Fayazi J, et al. (2011) Effects of different levels of zinc on the performance and carcass characteristics of broiler reared under heat stress condition. J Anim and Vet Adv 10(10), 1332-1335.
Salabi F, Nazari M, Chen Q, et al. (2014) Myostatin knockout using zinc-finger nucleases promotes proliferation of ovine primary satellite cells in vitro. J Biotech 192, 268-280.
SAS Institute (2016) Statistical Analysis Software (SAS) User’s Guide Version 9.4. Cary, NC: SAS Institute, Inc.
Sellier P (2000) Genetically caused retarded growth in animals. Domest Anim Endocrin 19, 105-119.
Seyedabadi HR, Asgari Jafarabadi Gh, Mahmoodi AR, Javaheri Barforosh H (2020) Effect of using flaxseed as omega-3 source on mammary gland growth and development gene expression in Saanen goat at their first pregnancy and lactation. Iranian Vet J 133, 32-44 (In Persian).
Shahsavari M, Mohammadabadi M, Khezri A, et al. (2023) Effect of Fennel (Foeniculum Vulgare) Seed Powder Consumption on Insulin-like Growth Factor 1 Gene Expression in the Liver Tissue of Growing Lambs. Gene Expr 21, 21-26.
Shokri S, Khezri A, Mohammadabadi M, Kheyrodin H (2023) The expression of MYH7 gene in femur, humeral muscle and back muscle tissues of fattening lambs of the Kermani breed. Agric Biotech J 15 (2), 217-236.
Spears JW (1996) Organic trace minerals in ruminant nutrition. Anim Feed Sci and Techno. 58, 151-163.
Starks TP (2006) Focus on Nutrition Research. Nova Publishers
Suttle NF (2010) The Mineral Nutrition of Livestock. CABI Publishing, New York.
Thissen JP, Ketelslegers JM, Underwood LE )1994) Nutritional regulation of the insulinlike growth factors. Endocr Rev 15, 80-101.
Tran LA, Malla BA, Sharma AN, et al. (2016) Effect of omega-3 and omega-6 polyunsaturated fatty acidenriched diet on plasma IGF-1 and testosteroneconcentration, puberty and semen quality in male buffalo. Anim Repro Sci 173, 63-72.‏
Turgut S, Kaptanoglu B, Emmungil G, Turgut G (2006) Increased plasma levels of growth hormone, insulin-like growth factor (IGF)-I and IGF-binding protein 3 in pregnant rats with exercise. Tohoku J Exp Med 208, 75e81.
Wei H, Zhou Y, Jiang S, et al. (2016) Transcriptional response of porcine skeletal muscle to feeding a linseed-enriched diet to growing pigs. J Anim Sci Biotech 7, 6.
Woods VB, Fearon AM (2009) Dietary sources of unsaturated fatty acids for animals and their transfer into meat, milk and eggs: A review. Livest Sci 126(1-3), 1-20.‏
Yu ZP, Le GW, Shi YH (2005) Effect of zinc sulphate and zinc methionine on growth, plasma growth hormone concentration, growth hormone receptor and insulin-like growth factor-I gene expression in mice. Clin and Exper Pharma and Physio 32, 273–278.