Yu Chengcheng, Wu Yinan, Huang Hanxu, Li Xiumao, Wang Jingkai, Liu Chao, Shen Yuanqing, Huang Donghua, Tang Ruofu, Wang Zhan, Jiang Lifeng, Li Fangcai
Department of Orthopedics, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Zhejiang Cancer Hospital, Institute of Medical Research, Chinese Academy of Sciences, Hangzhou, China.
Front Immunol. 2025 May 29;16:1550249. doi: 10.3389/fimmu.2025.1550249. eCollection 2025.
Traditional Chinese medicine and food deer antler has been extensively used in bone regeneration, but its molecular mechanisms remain poorly understood. Preliminary investigations suggest deer antler contains bioactive compounds that influence osteogenic differentiation and immune microenvironments.
To elucidate the molecular mechanisms of deer antler in promoting human mesenchymal stem cell (hMSC) osteogenic differentiation, focusing on JUN downregulation and immune microenvironment modulation using bioinformatics and molecular docking approaches.
Chemical components and targets were identified using the BATMAN-TCM database. Differentially expressed genes (DEGs) related to osteogenic differentiation were analyzed using Gene Expression Omnibus datasets. Gene Ontology (GO), KEGG enrichment, LASSO regression, and SVM-RFE were applied to identify key genes. A Protein-Protein Interaction (PPI) network was constructed to determine core genes. JUN expression was validated using independent datasets and ROC analysis. Immune cell infiltration was analyzed using CIBERSORT, examining JUN's correlation with immune cells. Molecular docking explored JUN's interaction with two active deer antler compounds.
The study identified 62 bioactive compounds and 1051 potential targets. DEGs analysis revealed 282 genes associated with osteogenic differentiation. Cross-analysis identified 43 overlapping genes, enriched in "response to mechanical stimulus" and "rheumatoid arthritis" pathways. Machine learning approaches highlighted 7 critical genes, with JUN emerging as the core gene. JUN levels were significantly decreased during osteogenic differentiation, showing high diagnostic accuracy (AUCs: 0.977-1.000). Immune cell analysis revealed JUN correlations with neutrophils, monocytes, eosinophils, M2 macrophages, and resting CD4+ T cells. Molecular docking confirmed strong binding affinities of JUN with Retinol (-8.1 kcal/mol) and Progesterone (-6.0 kcal/mol).
The study provides a comprehensive molecular framework demonstrating JUN as a key molecule in hMSC osteogenic differentiation. Deer antler's bioactive compounds, particularly Retinol and Progesterone, potentially exert therapeutic effects through targeted JUN modulation, offering novel insights into natural compound-mediated bone regenerative mechanisms.
中药和食药两用的鹿茸已广泛应用于骨再生，但对其分子机制仍知之甚少。初步研究表明，鹿茸含有影响成骨分化和免疫微环境的生物活性化合物。
运用生物信息学和分子对接方法，阐明鹿茸促进人间充质干细胞（hMSC）成骨分化的分子机制，重点关注JUN的下调和免疫微环境调节。
使用BATMAN-TCM数据库鉴定化学成分和靶点。利用基因表达综合数据集分析与成骨分化相关的差异表达基因（DEG）。应用基因本体论（GO）、KEGG富集、LASSO回归和支持向量机递归特征消除（SVM-RFE）来鉴定关键基因。构建蛋白质-蛋白质相互作用（PPI）网络以确定核心基因。使用独立数据集和ROC分析验证JUN的表达。使用CIBERSORT分析免疫细胞浸润，研究JUN与免疫细胞的相关性。分子对接探索JUN与两种鹿茸活性化合物的相互作用。
该研究鉴定出62种生物活性化合物和1051个潜在靶点。DEG分析揭示了282个与成骨分化相关的基因。交叉分析确定了43个重叠基因，富集于“对机械刺激的反应”和“类风湿性关节炎”途径。机器学习方法突出了7个关键基因，其中JUN成为核心基因。在成骨分化过程中，JUN水平显著降低，显示出较高的诊断准确性（曲线下面积：0.977 - 1.000）。免疫细胞分析揭示了JUN与中性粒细胞、单核细胞、嗜酸性粒细胞、M2巨噬细胞和静息CD4 + T细胞的相关性。分子对接证实JUN与视黄醇（-8.1千卡/摩尔）和孕酮（-6.0千卡/摩尔）具有很强的结合亲和力。
该研究提供了一个全面的分子框架，证明JUN是hMSC成骨分化中的关键分子。鹿茸的生物活性化合物，特别是视黄醇和孕酮，可能通过靶向调节JUN发挥治疗作用，为天然化合物介导的骨再生机制提供了新的见解。
