Zhejiang Guangsha Vocational and Technical University of Construction, Dongyang, People's Republic of China.
Zhejiang University of Technology, Hangzhou, People's Republic of China.
FASEB J. 2024 Oct 15;38(19):e70074. doi: 10.1096/fj.202401452R.
Diabetes osteoporosis (DOP) is a chronic metabolic bone disease. This study aimed to identify potential biomarkers of DOP and explore their underlying mechanisms through bioinformatics methods and experimental verification. Bioinformatics methods were used to identify differentially expressed genes (DEGs) for DOP based on GEO data and the GeneCards database. GO and KEGG enrichment analyses were used to search the key pathways. The STRING website was used to construct a protein-protein interaction (PPI) network and identify key genes. Then, 50 mg/mL glucose was used to interveneosteoblasts (OBs).CCK-8 and Alizarin Red staining were used to investigate the proliferation and differentiation changes in OBs. Flowcytometry was used to investigate apoptosis. The membrane protein chip, WB, and RT-PCR were used to verify the expression of key targets or pathways about DOP. Forty-two common genes were screened between DOP-related targets and DEGs. GO and KEGG enrichment analysis showed that DOP was mainly associated with cytokine-cytokine receptor interactions, and apoptosis. PPI network analysis showed that TNF, IL1A, IL6, IL1B, IL2RA, Fas ligand (FASLG), and Fas cell surface death receptor (FAS) were key up-regulated genes in the occurrence of DOP. The experiment results show that 50 mg/mL glucose significantly inhibited OBs proliferation but presented an increase in apoptosis. Membrane protein chip, WB, and RT-PCR-verified a significantly active in the expression of TNF/FASLG/FAS pathway. High glucose activated the TNF-α/FAS/FASLG pathway and induced the inflammatory microenvironment and apoptosis, then impaired osteogenic differentiation of OBs. These may be an important mechanism for the occurrence and development of DOP.
糖尿病性骨质疏松症（DOP）是一种慢性代谢性骨病。本研究旨在通过生物信息学方法和实验验证来鉴定 DOP 的潜在生物标志物，并探讨其潜在机制。基于 GEO 数据和基因卡片数据库，使用生物信息学方法鉴定 DOP 的差异表达基因（DEGs）。GO 和 KEGG 富集分析用于搜索关键途径。STRING 网站用于构建蛋白质-蛋白质相互作用（PPI）网络并识别关键基因。然后，用 50mg/mL 葡萄糖干预成骨细胞（OBs）。CCK-8 和茜素红染色用于研究 OBs 增殖和分化的变化。流式细胞术用于研究细胞凋亡。膜蛋白芯片、WB 和 RT-PCR 用于验证与 DOP 相关的关键靶标或途径的表达。在 DOP 相关靶标和 DEGs 之间筛选出 42 个共同基因。GO 和 KEGG 富集分析表明，DOP 主要与细胞因子-细胞因子受体相互作用和细胞凋亡有关。PPI 网络分析表明，TNF、IL1A、IL6、IL1B、IL2RA、Fas 配体（FASLG）和 Fas 细胞表面死亡受体（FAS）是 DOP 发生的关键上调基因。实验结果表明，50mg/mL 葡萄糖显著抑制 OBs 的增殖，但促进细胞凋亡。膜蛋白芯片、WB 和 RT-PCR 验证 TNF/FASLG/FAS 途径的表达明显活跃。高葡萄糖激活 TNF-α/FAS/FASLG 途径，诱导炎症微环境和细胞凋亡，进而损害 OBs 的成骨分化。这些可能是 DOP 发生和发展的重要机制。
