【目的】 蜜蜂是维持生态平衡的关键传粉昆虫，营养匮乏已成为其种群衰退的重要诱因之一。本研究旨在探究3种不同蛋白质日粮对意大利蜜蜂Apis mellifera ligustica高产浆蜂工蜂血淋巴代谢物与蛋白质表达的影响，从分子层面解析蛋白源差异对蜜蜂生理代谢的调控作用，为蜜蜂精准饲养提供理论依据。【方法】 在笼养条件下，将新出房工蜂分别饲喂茶花蜂花粉、油菜蜂花粉、大豆粉及无蛋白对照日粮，于9日龄时采集血淋巴，采用超高效液相色谱-质谱联用技术进行非靶向代谢组学与蛋白质组学分析，系统比较各组代谢物与蛋白质表达谱的差异。【结果】 代谢组学分析共鉴定出129种代谢物，与对照组相比，各日粮组均显著激活了苯丙氨酸，酪氨酸与色氨酸生物合成通路（P < 0.05）。相较于大豆粉，油菜蜂花粉饲喂显著上调了α-亚麻酸代谢通路（P < 0.05）。蛋白质组学分析共鉴定和定量到640种蛋白质，各日粮组普遍富集于碳代谢、氨基酸生物合成及糖酵解/糖异生等核心代谢途径，蜂花粉组较大豆粉组协同调控了α-亚麻酸代谢与花生四烯酸代谢通路（P < 0.05）。【结论】 补充蛋白质日粮可协同激活工蜂血淋巴中能量代谢及氨基酸合成的核心通路，从而有效支持其哺育期的蜂王浆分泌功能。相较于大豆粉，蜂花粉在脂质代谢和氨基酸生物合成等关键生理过程中展现出更全面、系统的营养支持作用。尤其值得注意的是，油菜蜂花粉在必需脂肪酸代谢、脂质合成与细胞膜功能调控方面表现出更显著的营养生理优势与代谢调控潜力。本研究结果为蜂群精准饲养与高效人工饲料的研发提供了重要依据。

  [Aim]  To elucidate the molecular mechanisms underlying diet-induced physiological regulation in a royal jelly-producing strain of honeybee (Apis mellifera ligustica), and thereby improve the husbandry of this strain. [Methods]  The effects of three different diets on the hemolymph metabolome and proteome of the royal jelly-producing strain were investigated. Caged newly emerged worker bees were randomly assigned to one of four dietary treatments: Camellia bee pollen, rapeseed bee pollen, soybean meal, or a protein-free, control diet. Hemolymph samples were collected from 9-day-old bees and analyzed using ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS)-based, untargeted metabolomics and proteomics, to systematically compare metabolite and protein profiles among the treatment groups. [Results]  A total of 129 metabolites were identified. Compared to the control group, all treatment groups had significantly higher phenylalanine, tyrosine and tryptophan biosynthesis, activity (P < 0.05). Notably, the α‑linolenic acid metabolism was markedly upregulated in the rapeseed bee pollen-fed group relative to the soybean meal-fed group (P < 0.05). A total of 640 proteins were identified and quantified by proteomics analysis. All dietary groups showed common enrichment in core metabolic pathways, including carbon metabolism, amino acid biosynthesis and glycolysis/gluconeogenesis. Furthermore, the bee pollen‑fed groups had stronger co‑regulation of the α‑linolenic acid and arachidonic acid metabolism than the soybean meal-fed group (P < 0.05). [Conclusion]  Protein supplementation enhances pathways related to energy metabolism and amino acid synthesis in worker bee hemolymph, thereby supporting royal jelly secretion in nurse bees. Among the protein sources evaluated, bee pollen provides superior integrated nutritional support than soybean meal, as demonstrated by the stronger enrichment of the lipid-metabolic and amino-acid biosynthetic pathways of bee-pollen fed bees. Notably, feeding bees rapeseed bee pollen resulted in greater modulation of essential fatty acid metabolism, lipid synthesis, and membrane-associated functions, indicating its suitability for precise adjustment of honeybee nutrition. Collectively, these findings provide a molecular basis for optimizing precision feeding strategies and formulating better artificial diets for honeybees.