This study focused on Stichopus monotuberculatus and conducted stress experiments at salinity levels of 20‰ and 40‰. Intestinal histological changes and the structural characteristics of the intestinal flora of S. monotuberculatus under salinity stress were analyzed. The results show that acute salinity stress inflicts varying degrees of damage to the intestinal tissues of S. monotuberculatus. Salinity stress enhances the species diversity of intestinal flora in S. monotuberculatus. Eight phyla of bacteria are detected in the intestine of S. monotuberculatus. Dominant phyla include Proteobacteria, Firmicutes, and Actinobacteria. Furthermore, functional prediction reveals that acute salinity stress can significantly modify the abundance of pathways associated with nutrient and energy metabolism mediated by the intestinal flora of S. monotuberculatus. These results indicate that acute salinity stress induces pathological damage to the intestinal tissues of S. monotuberculatus, compromising the microbial habitat and leading to alterations in the intestinal flora composition. Additionally, S. monotuberculatus can mitigate salinity stress by adjusting the composition of its intestinal flora and the corresponding functional pathways.

As a traditional element of Chinese medicine, Hippocampus erectus is well known for promoting adolescent growth, yet its active fractions and underlying molecular mechanisms remain unclear. In this study, the aqueous extract of H. erectus was subjected to in vitro simulated gastrointestinal digestion and ultrafiltration to separate three molecular weight fractions (30 kDa). Their chemical profiles were characterized, and osteogenic activities were systematically evaluated using cell assays, a juvenile rat model, and integrated transcriptomics and data-independent acquisition (DIA) proteomics. Results revealed that chemical profiling showed the >30 kDa fraction was mainly composed of hemocyanin subunits, and the 10–30 kDa fraction was enriched in growth-related amino acids and steroid derivatives; functionally, the 10–30 kDa fraction promoted preosteoblast proliferation and early differentiation via enhanced alkaline phosphatase (ALP) activity, while the >30 kDa fraction dominated late osteoblast maturation and mineralization. Both fractions significantly increased rat body and bone length by expanding growth plate proliferative zones and elevating serum insulin-like growth factor-1 (IGF-1)/bone morphogenetic protein-2 (BMP-2) levels. Transcriptomic and proteomic analyses identified vascular endothelial growth factor (VEGF), Wingless-related integration site (Wnt), phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt), and extracellular matrix (ECM)–receptor interaction as potential core regulatory pathways. Integrated multi-omics analysis further confirmed Frizzled-related protein B (Frzb) and AKT1 substrate 1 (Akt1s1) as candidate key regulatory targets enriched in the Wnt and adenosine monophosphate-activated protein kinase (AMPK) signaling pathways. These findings elucidate the multi-fraction, multi-pathway mechanism of H. erectus in promoting skeletal development, providing scientific evidence for its traditional use and a theoretical basis for growth-promoting functional food development.