Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 17 1 2025 03 21 Effect of zinc-methionine organic and Ca-salt of flaxseed oil supplement on IGF-1 gene expression in liver tissue of fattening male lambs Effect of zinc-methionine organic and Ca-salt of flaxseed oil supplement on IGF-1 gene expression in liver tissue of fattening male lambs 121 140 4736 10.22103/jab.2025.24414.1632 FA Mahmood Nazari Associate Professor, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Mollasani, Iran. Zeinab Alipoor Ph. D student, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Mollasani, Iran. 0009-0001-2715-497X Sepideh Rostami Ph. D student, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Mollasani, Iran. Ghazal Mohamadi Ahvazi Ph. D student, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Mollasani, Iran. Journal Article 2024 11 25 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. 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.

https://jab.uk.ac.ir/article_4736_622e47337f37a9e0600e68623e7420f8.pdf