Department of Clinical Laboratory, Shandong Provincial Hospital, Shandong University, Jinan, China.
Department of Reproductive Medicine, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, China.
PeerJ. 2023 Mar 3;11:e15041. doi: 10.7717/peerj.15041. eCollection 2023.
The elevating osteoclast differentiation can lead to an imbalance in bone homeostasis, which was responsible for bone loss and bone diseases, such as osteoporosis. Multiple pathways and molecules have been involved in osteoclast formation, but the role of CYP27A1 in osteoclast differentiation has never been explored.
CYP27A1 deficient mice were constructed using CRISPR-Cas9 system. Osteoclast differentiation was detected by TRAP staining. Differentially expressed genes (DEGs) were identified using RNA-seq analysis and were confirmed by qRT-PCR and Western blot.
The results showed that CYP27A1 knockout (KO) promoted osteoclast differentiation and bone loss. The transcriptomic analysis revealed that CYP27A1 KO led to differential expression of multiple genes, including ELANE, LY6C2, S100A9, GM20708, BGN, SPARC, and COL1A2, which were confirmed by qRT-PCR and Western blot. Enrichment analysis indicated that these differential genes were significantly associated with osteogenesis-related pathways, such as PPAR signaling, IL-17 signaling, and PI3K/AKT signaling, which were confirmed by qRT-PCR and Western blot.
These results suggested that CYP27A1 was involved in osteoclast differentiation, providing a novel therapeutic target for osteoclast-related diseases.
破骨细胞分化的增加会导致骨内环境失衡，从而导致骨质流失和骨疾病，如骨质疏松症。有多种途径和分子参与破骨细胞的形成，但 CYP27A1 在破骨细胞分化中的作用尚未被探索。
使用 CRISPR-Cas9 系统构建 CYP27A1 缺陷型小鼠。通过 TRAP 染色检测破骨细胞分化。使用 RNA-seq 分析鉴定差异表达基因（DEGs），并通过 qRT-PCR 和 Western blot 进行验证。
结果表明，CYP27A1 敲除（KO）促进了破骨细胞分化和骨质流失。转录组分析显示，CYP27A1 KO 导致多个基因的差异表达，包括 ELANE、LY6C2、S100A9、GM20708、BGN、SPARC 和 COL1A2，这些基因通过 qRT-PCR 和 Western blot 得到了验证。富集分析表明，这些差异基因与成骨相关途径显著相关，如 PPAR 信号、IL-17 信号和 PI3K/AKT 信号，这些途径通过 qRT-PCR 和 Western blot 得到了验证。
这些结果表明，CYP27A1 参与破骨细胞分化，为破骨细胞相关疾病提供了一个新的治疗靶点。
