Effects of Dietary Vitamin D3 Supplementation on Growth Performance, Immune and Antioxidant Functions in the Adipose Tissue of Luhua Broilers (Gallus gallus)
TAN Li-You, WU Jia-Hui, LI Jia-Wei, WANG Ya-Peng, CHEN Yan, DAI Hong-Wei, LU Jian-Xiong, JIANG Su-Su*, ZHANG Guo-Hua*
School of Life Sciences and Engineering, Northwest Minzu University, Lanzhou 730124, China
Abstract:Vitamin D3 (VD3) has various biological functions. Beyond regulating the absorption and utilization of calcium and phosphorus, it also plays a significant role in the metabolism and overall health of animals. A total of 210 one-day-old male Luhua broilers (Gallus gallus) were randomly assigned to a control (basal diet) and VD3 group (basal diet supplemented with 3750 IU/kg VD3), with 7 replicates per group in this study. The 84-day experiment was conducted to investigate the effects of dietary vitamin D3 supplementation on growth performance, immunity and antioxidant function of Luhua broilers, by determining the relative weight of immune organs, as well as immune factors and antioxidant indexes in serum and adipose tissue. The results showed that dietary VD3 supplementation significantly increased body weight gain of broilers aged from 1 to 84 d (P<0.05). Supplementary VD3 significantly increased the serum 25-hydroxyvitamin D3 (25(OH)D3) level and upregulated the mRNA expression of vitamin D receptor (VDR), cytochrome P450 family 2 subfamily R member 1 (CYP2R1), cytochrome P450 family 27 subfamily C member 1 (CYP27C1), cytochrome P450 family 24 subfamily A member 1 (CYP24A1), and vitamin D binding protein (VDBP) in the adipose tissue of broilers (P<0.05). Dietary VD3 significantly decreased serum malondialdehyde (MDA) levels and increased the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in the serum of broilers at 56 days of age, as well as GSH-Px and CAT activities at 84 days of age; additionally, VD3 significantly upregulated the mRNA expression of antioxidant-related genes, including nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NADPH quinone oxidoreductase 1 (NQO1), GSH-Px, and SOD in the adipose tissue of broilers at 56 and 84 days of age (P<0.05). Dietary VD3 significantly increased the organ indices of thymus and bursa of Fabricius in broilers at 56 and 84 days of age, and serum levels of immunoglobulin A (IgA), IgG, interleukin-2 (IL-2) and interferon-γ (IFN-γ) in broilers at 28, 56 and 84 d; significantly increased levels of IgA, IgG, IL-10 in adipose tissue at all ages, as well as IgM levels at 56 and 84 days of age. It also significantly downregulated the mRNA expression of tumor necrosis factor-α (TNF-α), nuclear factor kappa-B (NF-κB), toll-like receptor-4 (TLR-4) and IL-1β, and upregulated mRNA expression of IL-10 in the adipose tissue (P<0.05). These results indicated that dietary supplementation with 3 750 IU/kg VD3 could enhance VD3 metabolism and improve systemic and adipose tissue antioxidant and immune functions, thereby supporting the health and growth of Luhua broilers. The findings of this study provide a basis for a deeper understanding of the nutritional and physiological functions of VD3 and for optimizing its utilization in broiler production.
[1] 从光雷, 肖蕴祺, 张倩雲, 等. 2021. 饲粮添加橡椀单宁对肉鸡生长性能、免疫器官指数和血浆生化指标的影响[J]. 动物营养学报, 33(5): 2652-2660. (Cong G L, Xiao Y Q, Zhang Q Y, et al.2021. Effects of dietary quercus acutissima carruth tannin on growth performance, immune organ indexes and plasma biochemical indexes of broilers[J]. Chinese Journal of Animal Nutrition, 33(5):2652-2660.) [2] 符璐, 苗健, 张国华, 等. 2023. 维生素D调控动物脂肪形成及脂肪组织代谢的研究进展[J]. 动物营养学报, 35(9): 5545-5554. (Fu L, Miao J, Zhang G H, et al.2023. Research advances in regulation of animal adipogenesis and adipose tissue metabolism by vitamin D[J]. Chinese Journal of Animal Nutrition, 35(09): 5545-5554.) [3] 韩涵. 2019. 维生素D3通过NF-κB/PPARγ通路对2型糖尿病大鼠脂肪组织炎症的影响[D]. 硕士学位论文, 郑州大学, 导师: 李文杰, pp. 38-45. (Han H.2019. The effect of vitamin D3 on adipose inflammation in type 2 diabetic rats through NF-κB/PPARγ pathway[D]. Thesis for M. S., Zhengzhou University, Supervisor: Li W J, pp. 38-45.) [4] 李思明. 2009. 维生素D3对丝毛鸡β-防御素诱导表达及生长免疫性能的影响[D]. 博士学位论文, 四川农业大学, 导师: 周定刚, pp. 117-118. (Li S M.2009. Effects of vitamin D3 on growth performance and induction of the β-defensins expression in silky fowl[D]. Thesis for Ph.D., Sichuan Agricultural University, Supervisor: Zhou D G, pp.117-118.) [5] 南瑛, 赵美娜, 张薇. 2018. α-亚麻酸抑制高脂诱导的脂肪细胞氧化应激和促炎因子的释放[J]. 免疫学杂志, 34(11): 921-927. (Nan Y, Zhao M N, Zhang W.2018. ALA inhibits oxidative stress and the generation of proinflammatory cytokines induced by free fatty acids in adipocyte[J]. Immunological Journal, 34(11): 921-927.) [6] 欧阳怡佩. 2024. 黄根多糖通过Nrf2/NQO1/HO-1抗氧化通路对肺纤维化的抑制作用[D]. 硕士学位论文, 右江民族医学院, 导师: 黄世稳, pp. 50-55. (Ou Yang Y P.2024. Inhibitory effect of PSPT on pulmonary fibrosis via the Nrf2/NQO1/HO-1 antioxidant pathway[D]. Thesis for M. S., Youjiang Medical University For Nationalities, Supervisor: Huang S W, pp. 50-55.) [7] 尚书凤, 赵小龙, 杨书洋, 等. 2022. 1α羟化酶的真核表达和生物信息学特征及其在成年肉鸡组织中的差异表达[J]. 江苏农业学报, 38(05): 1286-1297. (Shang S F, Zhao X L, Yang S Yet al.2022. Eukaryotic expression and bioinformatics characteristics of 1α hydroxylase and its differential expression in adult broiler tissues[J]. Jiangsu Journal of Agricultural Sciences, 38(05): 1286-1297.) [8] 王一冰, 张盛, 陈志龙, 等. 2022. 维生素D3对黄羽肉种鸡生殖性状及其后代肉鸡生长性能、免疫器官发育、胫骨性状和肉品质的影响[J]. 动物营养学报, 34(03): 1533-1546. (Wang Y B, Zhang S, Chen Z L, et al.2022. Effects of vitamin D3 on reproductive traits of yellow-feathered broiler breeders and growth performance, immune organ development, tibia characteristics and meat quality of offspring broilers[J]. Chinese Journal of Animal Nutrition, 34(03): 1533-1546.) [9] 张淑云, 王安. 2010. 钙和维生素D对生长肉鸡免疫及抗氧化功能的影响[J]. 动物营养学报, 22(03): 579-585. (Zhang S Y, Wang A.2010. Effects of calcium and vitamin D on immune and antioxidant functions in growing broilers[J]. Chinese Journal of Animal Nutrition, 22(03):579-585.) [10] 赵连鹏. 2021. 母猪妊娠期饲粮添加维生素D3对其繁殖性能及母仔猪抗氧化能力的影响[D]. 硕士学位论文, 四川农业大学, 导师: 冯斌, 罗何峰, pp. 31-36. (Zhao L P.2021. Effect of dietary vitamin D3 supplementation in sow during gestation on reproductive performance and antioxidant capacity of sows and piglets[D]. Thesis for M. S., Sichuan Agricultural University, Supervisor: Feng B, Luo H F, pp. 31-36.) [11] Almasmoum H, Refaat B, Ghaith M M, et al.2019. Protective effect of vitamin D3 against lead induced hepatotoxicity, oxidative stress, immunosuppressive and calcium homeostasis disorders in rat[J]. Environmental Toxicology and Pharmacology, 72: 103246. [12] Attia Y A, Al-Harthi M A, Abo H M.2020. Calcium and cholecalciferol levels in late-phase laying hens: Effects on productive traits, egg quality, blood biochemistry, and immune responses[J]. Frontiers in Veterinary Science, 7: 389. [13] Baker R G, Hayden M S, Sand Ghosh S.2011. NF-κB, inflammation, and metabolic disease[J]. Cell Metabolism, 13(1): 11-22. [14] Bonnet L, Karkeni E, Couturier C, et al.2017. Gene expression pattern in response to cholecalciferol supplementation highlights cubilin as a major protein of 25(OH)D uptake in adipocytes and male mice white adipose tissue[J]. Endocrinology, 159(2): 957-966. [15] Bscheider M, Butcher E C.2016. Vitamin D immunoregulation through dendritic cells[J]. Immunology, 148(3): 227-236. [16] Chang E.2022. Effects of vitamin D supplementation on sdipose tissue inflammation and NF-κB/AMPK activation in obese mice fed a high-fat diet[J]. International Journal of Molecular Sciences, 23(18): 10915. [17] Chen L, Yang R, Qiao W, et al.2019. 1,25-Dihydroxyvitamin D exerts an antiaging role by activation of Nrf2-antioxidant signaling and inactivation of p16/p53-senescence signaling[J]. Aging Cell, 18(3): e12951. [18] Chen X, Zeng D, Zeng X, et al.2024. Effects of complex antioxidants added to chicken diet on growth performance, serum biochemical indices, meat quality, and antioxidant capacity[J]. Animals, 14(3): 360. [19] Chun R F, Liu P T, Modlin R L, et al.2014. Impact of vitamin D on immune function: Lessons learned from genome-wide analysis[J]. Frontiers in Physiology, 5: 151. [20] Fan W J, Tang T T, Qian Z, et al.2020. The role of IL-1β in aortic aneurysm[J]. Clinica Chimica Acta, 504: 7-14. [21] Fatemi S A, Macklin K S, Zhang L, et al.2022. Improvement in the immunity- and vitamin D3-activity-related gene expression of coccidiosis-challenged ross 708 broilers in response to the in ovo injection of 25-hydroxyvitamin D3[J]. Animals (Basel), 12(19): 2517. [22] Fiore N T, Willcox K F, Dayani D, et al.2025. Reducing IgG accumulation via neonatal Fc receptor (FcRn) blockade relieves neuropathic pain[J]. Brain, Behavior, and Immunity, 125: 371-387. [23] Gao H, Wang Y, Zhao X, et al.2025. Growth performance and gut health of cold-stressed broilers in response to supplementation with a combination of sodium butyrate and Vitamin D3[J]. Animals (Basel), 15(6): 861. [24] He J, Wang H, Shi J, et al.2021. 1,25-dihydroxyvitamin D deficiency accelerates male reproductive senescence in aging mice and 1,25(OH)(2)D(3) alleviates oxidative stress via NF-κB/SOD[J]. American Journal of Physiology Endocrinology and Metabolism, 320(4): 732-746. [25] Hernandez R, Põder J, LaPorte K M, et al.2022. Engineering IL-2 for immunotherapy of autoimmunity and cancer[J]. Nature Reviews Immunology, 22(10): 614-628. [26] Hui L, Jie X, Qian S, et al.2024. Active vitamin D ameliorates arsenite-induced thyroid dysfunction in sprague-dawley rats by inhibiting the Toll-like receptor 4/NF-KappaB-mediated inflammatory response[J]. Toxics, 12(12): 887. [27] Karkeni E, Bonnet L, Marcotorchino J, et al.2018. Vitamin D limits inflammation-linked microRNA expression in adipocytes in vitro and in vivo: A new mechanism for the regulation of inflammation by vitamin D[J]. Epigenetics, 13(2): 156-162. [28] Karkeni E, Marcotorchino J, Tourniaire F, et al.2015. Vitamin D limits chemokine expression in adipocytes and macrophage migration in vitro and in male mice[J]. Endocrinology, 156(5): 1782-1793. [29] Katarína K, Viktória D, Kristína F, et al.2024. Modification of adipogenesis and oxidative stress by quercetin: positive or negative impact on adipose tissue metabolism of obese diabetic zucker rats?[J]. Journal of Physiology and Biochemistry, 81(1): 137-156. [30] Krawiec P, Pawłowska K A, Pac K E.2021. Interleukin 10 and interleukin 10 receptor in paediatric inflammatory bowel disease: From bench to bedside lesson[J]. Journal of Inflammation, 18(1): 13. [31] Kveler K, Starosvetsky E, Ziv K A, et al.2018. Immune-centric network of cytokines and cells in disease context identified by computational mining of pubmed[J]. Nature Biotechnology, 36(7): 651-659. [32] Li J, Bi Q, Pi Y, et al.2024b. Dietary supplementation with 25-hydroxyvitamin D3 on reproductive performance and placental oxidative stress in primiparous sows during mid-to-late gestation[J]. Antioxidants, 13(9): 1090. [33] Li J W, Li X M, Tian J M, et al.2025. Effects of supplementation with vitamin D(3) on growth performance, lipid metabolism and cecal microbiota in broiler chickens[J]. Frontiers in Veterinary Science, 12: 1542637. [34] Li L, Han B, Kong Y, et al.2024a. Vitamin D binding protein in psychiatric and neurological disorders: Implications for diagnosis and treatment[J]. Genes & Diseases, 11(5): 101309. [35] Liu X.2025. The paradoxical role of IFN-γ in cancer: Balancing immune activation and immune evasion[J]. Pathology, Research and Practice, 272: 156046. [36] Lv L, Tan X, Peng X, et al.2020. The relationships of vitamin D, vitamin D receptor gene polymorphisms, and vitamin D supplementation with Parkinson's disease[J]. Translational Neurodegeneration, 9(1): 34. [37] Miao Y F, Jiang Z G, Song H L, et al.2024. Vitamin D supplementation alleviates high fat diet-induced metabolic associated fatty liver disease by inhibiting ferroptosis pathway[J]. European Journal of Nutrition, 64(1): 50. [38] Mirza I, Mohamed A, Deen H, et al.2022. Obesity-associated vitamin D deficiency correlates with adipose tissue DNA hypomethylation, inflammation, and vascular dysfunction[J]. International Journal of Molecular Sciences, 23(22): 14377. [39] Mukherjee T, Kumar N, Chawla M, et al.2024. The NF-κB signaling system in the immunopathogenesis of inflammatory bowel disease[J]. Science Signaling, 17(818): eadh1641. [40] Palhinha L, Liechocki S, Hottz E D, et al.2019. Leptin induces proadipogenic and proinflammatory signaling in adipocytes[J]. Frontiers in Endocrinology, 10: 841. [41] Palming J, Gabrielsson G B, Eva J, et al.2006. Plasma cells and Fc receptors in human adipose tissue--lipogenic and anti-inflammatory effects of immunoglobulins on adipocytes[J]. Biochemical and Biophysical Research Communications, 343(1): 43-48. [42] Phutthaphol R, Bunchasak C, Loongyai W, et al.2025. Effects of vitamin D(3) and 25(OH)D(3) supplementation on growth performance, bone parameters and gut microbiota of broiler chickens[J]. Animals (Basel), 15(19): 2900. [43] Picatoste B M, Cerro P I, Garcia G A, et al.2024. Role of polymeric immunoglobulin receptor in glucose metabolism[J]. Cardiovascular Research, 120(1): 190. [44] Rajbhandari P, Thomas B J, Feng A C, et al.2017. IL-10 signaling remodels adipose chromatin architecture to limit thermogenesis and energy expenditure[J]. Cell, 172(1-2): 218-233.e217. [45] Randall T D, Meza-Perez S.2024. Immunity in adipose tissues: Cutting through the fat[J]. Immunological Reviews, 324(1): 4-10. [46] Rasheed A, Khan G.2024. Epstein-Barr virus, vitamin D and the immune response: Connections with consequences for multiple sclerosis[J]. Frontiers in Immunology, 15: 1503808. [47] Richter F C, Friedrich M, Kampschulte N, et al.2023. Adipocyte autophagy limits gut inflammation by controlling oxylipin and IL-10[J]. The EMBO Journal, 42(6): e112202. [48] Saponaro F, Saba A, Zucchi R.2020. An update on vitamin D metabolism[J]. International Journal of Molecular Sciences, 21(18): 6573. [49] Shadid I L C, Guchelaar H J, Weiss S T, et al.2024. Vitamin D beyond the blood: Tissue distribution of vitamin D metabolites after supplementation[J]. Life Sciences, 355: 122942. [50] Shinya S, Yohannes Y B, Ikenaka Y, et al.2022. Characteristics of cytochrome P450-dependent metabolism in the liver of the wild raccoon, procyon lotor[J]. The Journal of Veterinary Medical Science, 84(12): 1665-1672. [51] Shojadoost B, Behboudi S, Villanueva A I, et al.2015. Vitamin D3 modulates the function of chicken macrophages[J]. Research in Veterinary Science, 100: 45-51. [52] Sies H, Berndt C, Jones D P.2017. Oxidative stress[J]. Annual Review of Biochemistry, 86(1): 715-748. [53] Surai P F, Kochish I I.2019. Nutritional modulation of the antioxidant capacities in poultry: The case of selenium[J]. Poultry Science, 98(10): 4231-4239. [54] Suttorp M, Classen C F.2021. Splenomegaly in children and adolescents[J]. Frontiers in Pediatrics, 9: 704635. [55] Unamuno X, Gómez A J, Rodríguez A, et al.2018. Adipokine dysregulation and adipose tissue inflammation in human obesity[J]. European Journal of Clinical Investigation, 48(9): e12997. [56] Vázquez Lorente H, Herrera Q L, Jiménez S L, et al.2024. Antioxidant functions of vitamin D and CYP11A1-derived vitamin D, tachysterol, and lumisterol metabolites: Mechanisms, clinical implications, and future directions[J]. Antioxidants (Basel), 13(8): 996. [57] Wang Z, Zuo Z, Li L, et al.2020. Nrf2 in adipocytes[J]. Archives of Pharmacal Research, 43(3): 350-360. [58] Wei J, Li L, Peng Y, et al.2024. The Effects of optimal dietary vitamin D(3) on growth and carcass performance, tibia traits, meat quality, and intestinal morphology of chinese yellow-feathered broiler chickens[J]. Animals (Basel), 14(6): 920. [59] Wu H Z, Ballantyne C M.2020. Metabolic inflammation and insulin resistance in obesity[J]. Circulation Research, 126(11): 1549-1564. [60] Xiao C, Zhu Q, Comer L, et al.2023. Dietary 25-hydroxy-cholecalciferol and additional vitamin E improve bone development and antioxidant capacity in high-density stocking broilers[J]. Journal of Animal Science, 101: skad369. [61] Yamamoto M, Kensler T W, Motohashi H.2018. The KEAP1-NRF2 system: A thiol-based sensor-effector apparatus for maintaining redox homeostasis[J]. Physiological Reviews, 98(3): 1169-1203. [62] Yang J, Tian G, Chen D, et al.2019. Effects of dietary 25-hydroxyvitamin D(3) supplementation on growth performance, immune function and antioxidative capacity in weaned piglets[J]. Archives of Animal Nutrition, 73(1): 44-51. [63] Yi N K, Ming L Y, Xin F, et al.2023. Effects of the Vitamin D3 on alleviating the oxidative stress induced by diquat in wenchang chickens[J]. Animals, 13(4): 711. [64] Yu L X, Yang Y X, Gong Z, et al.2024. FcRn-dependent IgG accumulation in adipose tissue unmasks obesity pathophysiology[J]. Cell Metabolism, 37(3): 656-672. [65] Zhang Z, Zhang G, Cao J, et al.2022. Effects of 25(OH)VD3 on growth performance, pork quality and calcium deposit in growing-finishing pigs[J]. Animals, 13(1): 86. [66] Zhou X, Zou Y, Xu Y, et al.2022. Dietary supplementation of 25-hydroxyvitamin D(3) improves growth performance, antioxidant capacity and immune function in weaned piglets[J]. Antioxidants (Basel), 11(9): 1750.