Lactoferrin (LF) is believed to be an important active protein in goat milk, which plays an anti-inflammatory role. Although LF has been reported to be associated with body health, its exact underlying mechanism remains unclear. Here, we aimed to elucidate the mechanism of this anti-inflammatory effect of LF in vitro. We first identified that miR-214-5p inhibited the expression of LF mRNA and protein in cells through the 3′UTR of LF mRNA. We next identified the alterations in miRNA following LF overexpression in goat mammary epithelial cells (GEMCs). Overexpression of LF significantly increased (p < 0.05) miR-224-5p expression. We further revealed that transcriptional activation of ADAM17, TNF-α, IL-1β, and IL-6 was efficiently decreased (p < 0.05) in GMECs treated by miR-224-5p mimic. Conversely, knockdown of miR-224-5p increased (p < 0.05) ADAM17, TNF-α, IL-1β, and IL-6 expression. Additionally, TNF-α, IL-1β, and IL-6 expression levels were dramatically decreased in GMECs after administration of siADAM17. Herein, we indicate that the miR-214-5p/LF/miR-224-5p/ADAM17 axis is involved in the immune regulation of GEMCs.

Lentinus edodes (LE) is a nutrient-rich medicinal fungus with potential applications in animal nutrition; however, its effects on dairy goats remain underexplored. This study investigated the impact of dietary LE addition on rumen microbiota, metabolic profiles, serum immunity, and milk quality in lactating dairy goats. Twenty Saanen goats were randomly assigned to a control group (CON, basal diet) or an LE group (basal diet + 25 g Lentinus edodes) for 56 days. Rumen fluid, serum, and milk samples were analyzed using 16S rDNA sequencing, metabolomics, and biochemical assays. The results showed that the addition of LE altered the microbial composition, decreasing the abundance of fibrobacterial flora and Treponema (p < 0.05) while increasing VadinHA49. Metabolomic analysis revealed elevated fumaric acid, lysophospholipids (LysoPE, LysoPG), and D-quinic acid in the rumen (p < 0.05). Serum immunoglobulin A (IgA), immunoglobulin G (IgG), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), and tumor necrosis factor-α (TNF-α) levels were significantly enhanced (p < 0.05). The milk somatic cell count (SCC) was reduced by 52.8% (p = 0.005) at day 56, and protein, fat, and total solids increased significantly (p < 0.05). Enrichment of functional metabolites such as D-arabitol and D-tryptophan in milk further highlighted LE’s role in improving product value. These findings demonstrate that LE optimizes rumen flora, enhances antioxidant and immune functions, and improves milk quality, supporting its use as a functional feed additive for dairy goats.