Volume 15, Issue 1 (4-2024)
Res Anim Prod 2024, 15(1): 63-72 |
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Mirzaei H, Aghaei A, Hoveizi E, Nazari M. (2024). The Effect of Chromium Propionate and Alpha-Lipoic Acid Supplementation on Gene Expression of Cytokine Profile in the Spleen Tissue of Broilers. Res Anim Prod. 15(1), 63-72. doi:10.61186/rap.15.43.57
URL: http://rap.sanru.ac.ir/article-1-1345-en.html
URL: http://rap.sanru.ac.ir/article-1-1345-en.html
1- Department of Animal Science, Faculty of Animal Science and Food Technology, Agricultural sciences and Natural Resources University of Khuzestan, Iran
2- Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran
3- Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran
2- Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran
3- Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran
Abstract: (1343 Views)
Extended Abstract
Background: Alpha-lipoic acid (α-LA) is synthesized enzymatically in the mitochondrion from octanoic acid. Endogenous levels of α-lipoic acid have been reported in the micromolar range. Due to its high lipophilicity, alpha-lipoic acid can easily pass through biological membranes and is converted into dihydrolipoic acid (DHLA) in cells. Alpha-lipoic acid and DHLA are described as an ideal antioxidant couple due to their exceptional ability to protect against oxidative stress through multiple pathways. Both compounds exhibit metal chelating activity, act as coenzymes in glucose metabolism, and can produce other antioxidants such as ascorbate, vitamin E, and glutathione (GST). Alpha-lipoic acid also has anti-inflammatory properties, and its supplementation in broiler diets improves the expression of inflammation-related genes in the spleen.
On the other hand, trivalent chromium is recognized as an essential trace element for humans and other animals, as it is part of the glucose tolerance factor (GTF). Chromium is essential for the metabolism of carbohydrates, proteins, and lipids and can be present in the diet in the form of inorganic compounds or organic complexes. After absorption, chromium circulates in a free state and binds to transferrin or other plasma proteins (such as β-globulin). Trivalent chromium appears to play a role in the structure and expression of genetic information in animals, with stronger binding to nucleic acids than other metal ions. Chromium protects RNA against thermal denaturation and is concentrated in the cell nucleus, where it binds to chromatin, participates in gene expression, and increases the initiation sites for RNA synthesis. This increase results from the induction of proteins bound to the nucleus and the activation of nuclear chromatin, thereby affecting gene function. Chromium supplementation has shown a positive effect on the immune response of broiler chickens. The aim of this research was to investigate the simultaneous effects of dietary chromium propionate and alpha-lipoic acid supplementation on the expression of cytokines interleukin 1, 2, 4, and 10, as well as IFN-γ, TNF, and TGF-β4 genes in the spleen tissue of broiler chickens.
Methods: For this purpose, an experiment was conducted with 144 Ross 308 broiler chicks in a completely randomized design (CRD) with a 3×2 factorial arrangement of treatments, including six treatments, three replicates, and eight chickens in each replicate. The dietary treatments consisted of three doses of chromium propionate (0, 750, and 1500 μg/kg) combined with two levels of alpha-lipoic acid (0 and 300 mg/kg) supplemented to the basal diets. At the end of the experimental period (42 days old), two chickens from each pen were randomly selected and euthanized. Samples were taken from the spleen, immediately frozen in liquid nitrogen, and then stored at -80°C. The quantity and purity of the extracted RNA were checked using a Nanodrop device. To confirm the specific amplification of the desired genes, PCR reactions were performed using specific primers. The expression of cytokine profile genes was quantified by quantitative real-time PCR, using 28S as a housekeeping gene to normalize the gene expression data. The method of analyzing the obtained data employed the difference in threshold degree changes (∆∆CT), using the 2-ΔΔCT method (comparative threshold) and the ratio of gene expression (28S).
Results: The results of electrophoresis confirmed the expression of genes in the spleen cells of broiler chickens, appearing as one band on a 2% agarose gel. Electrophoresis of polymerase chain reaction products showed fragments of 86, 51, 82, and 88 bp for interleukin 1, 2, 4, and 10, respectively. No bands were formed on the agarose gel for the IFN-γ, TNF, and TGF-β4 genes. The results indicated that alpha-lipoic acid at the level of 300 mg/kg significantly decreased interleukin 1 gene expression compared to the control group (0 mg/kg). Additionally, supplementing the basal diet with chromium propionate at the level of 1500 μg/kg significantly increased interleukin 1 and 4 gene expression compared to the control group and the chromium propionate at the level of 750 μg/kg. However, the addition of alpha-lipoic acid and chromium propionate had no effect on the expression of interleukin 2 and 10 genes.
Conclusion: Adding 300 mg/kg of alpha-lipoic acid to the diet can improve the immune response by reducing the expression of the IL-1 gene in broilers. Furthermore, high amounts of chromium propionate (1500 μg) may lead to decreased immunity by increasing the expression of IL-1 and 4 genes. In contrast, the addition of chromium propionate up to the level of 750 μg/kg does not adversely affect immunity. Based on the results, it is recommended to include alpha-lipoic acid at the level of 300 mg/kg in the broiler diet, while the addition of chromium propionate up to 750 μg/kg is acceptable if it has beneficial effects on yield and production characteristics.
On the other hand, trivalent chromium is recognized as an essential trace element for humans and other animals, as it is part of the glucose tolerance factor (GTF). Chromium is essential for the metabolism of carbohydrates, proteins, and lipids and can be present in the diet in the form of inorganic compounds or organic complexes. After absorption, chromium circulates in a free state and binds to transferrin or other plasma proteins (such as β-globulin). Trivalent chromium appears to play a role in the structure and expression of genetic information in animals, with stronger binding to nucleic acids than other metal ions. Chromium protects RNA against thermal denaturation and is concentrated in the cell nucleus, where it binds to chromatin, participates in gene expression, and increases the initiation sites for RNA synthesis. This increase results from the induction of proteins bound to the nucleus and the activation of nuclear chromatin, thereby affecting gene function. Chromium supplementation has shown a positive effect on the immune response of broiler chickens. The aim of this research was to investigate the simultaneous effects of dietary chromium propionate and alpha-lipoic acid supplementation on the expression of cytokines interleukin 1, 2, 4, and 10, as well as IFN-γ, TNF, and TGF-β4 genes in the spleen tissue of broiler chickens.
Methods: For this purpose, an experiment was conducted with 144 Ross 308 broiler chicks in a completely randomized design (CRD) with a 3×2 factorial arrangement of treatments, including six treatments, three replicates, and eight chickens in each replicate. The dietary treatments consisted of three doses of chromium propionate (0, 750, and 1500 μg/kg) combined with two levels of alpha-lipoic acid (0 and 300 mg/kg) supplemented to the basal diets. At the end of the experimental period (42 days old), two chickens from each pen were randomly selected and euthanized. Samples were taken from the spleen, immediately frozen in liquid nitrogen, and then stored at -80°C. The quantity and purity of the extracted RNA were checked using a Nanodrop device. To confirm the specific amplification of the desired genes, PCR reactions were performed using specific primers. The expression of cytokine profile genes was quantified by quantitative real-time PCR, using 28S as a housekeeping gene to normalize the gene expression data. The method of analyzing the obtained data employed the difference in threshold degree changes (∆∆CT), using the 2-ΔΔCT method (comparative threshold) and the ratio of gene expression (28S).
Results: The results of electrophoresis confirmed the expression of genes in the spleen cells of broiler chickens, appearing as one band on a 2% agarose gel. Electrophoresis of polymerase chain reaction products showed fragments of 86, 51, 82, and 88 bp for interleukin 1, 2, 4, and 10, respectively. No bands were formed on the agarose gel for the IFN-γ, TNF, and TGF-β4 genes. The results indicated that alpha-lipoic acid at the level of 300 mg/kg significantly decreased interleukin 1 gene expression compared to the control group (0 mg/kg). Additionally, supplementing the basal diet with chromium propionate at the level of 1500 μg/kg significantly increased interleukin 1 and 4 gene expression compared to the control group and the chromium propionate at the level of 750 μg/kg. However, the addition of alpha-lipoic acid and chromium propionate had no effect on the expression of interleukin 2 and 10 genes.
Conclusion: Adding 300 mg/kg of alpha-lipoic acid to the diet can improve the immune response by reducing the expression of the IL-1 gene in broilers. Furthermore, high amounts of chromium propionate (1500 μg) may lead to decreased immunity by increasing the expression of IL-1 and 4 genes. In contrast, the addition of chromium propionate up to the level of 750 μg/kg does not adversely affect immunity. Based on the results, it is recommended to include alpha-lipoic acid at the level of 300 mg/kg in the broiler diet, while the addition of chromium propionate up to 750 μg/kg is acceptable if it has beneficial effects on yield and production characteristics.
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