Study on the effect of deer bone in improving rheumatoid arthritis based on the "drug-target-pathway" association network

Ethnopharmacological relevance: Deer bone is rich in proteins, free Amino acids, chondroitin, organic calcium, phosphorus ions, and Other active components. Deer bone had been used widely in antiquity and were first compiled in renowned ancient masterpiece 'Mingyi Bielu ()' written by Hongjing Tao. The deer bone is recorded as non-toxic and has the effects of replenishing bones, strengthening sinews, expelling wind-dampness from the body, promoting muscle growth, and healing wounds. Modern pharmacological research suggests that deer bone can help promote bone density and enhance bone strength, making it potentially valuable for the prevention and treatment of diseases such as rheumatoid arthritis and osteoporosis. However, current studies on the component analysis and pharmacological effects of deer bone against rheumatoid arthritis (RA) are incomplete, which to some extent hinders the development and clinical application of deer bone drugs.

Aim of the study: The components of deer bone were elucidated by label-free proteomics, and the drug-target-pathway association network was established by network pharmacology. The in vitro validation of the pathway provides a theoretical basis for deer bone as a potential therapeutic drug for rheumatoid arthritis, and also lays a solid foundation for the subsequent clinical application of the in vitro experiments established through serum pharmacology.

Materials and methods: We performed extraction of deer bone using traditional water extraction methods and employed label-free proteomics technology to identify and conduct bioinformatics analysis on the proteins and peptides in the deer bone hot water extract (DBHE). These components were considered potential drug targets, and we constructed a "drug-target-pathway" association network. Analysis revealed that the HIF-1 signaling pathway may be pivotal in DBWE's effect on RA. Hypoxia influences the occurrence and development of Ferroptosis through various mechanisms. Therefore, we hypothesized that DBWE might induce Ferroptosis, promoting Apoptosis in RA-FLS under hypoxic conditions, thereby alleviating RA. Therefore, we performed flow cytometry, ELISA, immunofluorescence, RT-qPCR, and western blotting based on molecular docking. Considering the overall effect of drug metabolism post-ingestion, we used serum pharmacology to prepare serum for cellular administration.

Results: It showed that DBWE reduces inflammation and synovial proliferation by inhibiting HO-1, increasing ROS production, upregulating ACSL4 expression and inducing RA-FLS Apoptosis in hypoxic conditions. This study reveals the potential mechanism by which DBWE modulates Ferroptosis to attenuate synovial proliferation in a hypoxic microenvironment and improve RA.

Conclusion: These findings not only provide a theoretical basis for deer bone as a potential therapeutic agent for RA, but also lay a solid foundation for subsequent clinical application through in vitro experiments established by serum pharmacology.

Deer bone; Molecular docking; Network pharmacology; Rheumatoid arthritis; Synovial hypoxia.