Lu Xin, Fan Yu, Li Mingxia, Chang Xiao, Qian Jun
Department of Orthopaedics, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
The Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Front Mol Biosci. 2021 Jun 16;8:691602. doi: 10.3389/fmolb.2021.691602. eCollection 2021.
Osteoarthritis (OA) is a heterogeneous age-related disease, which is badly difficult to cure due to its complex regulatory networks of pathogenesis. This study explored OA-specific genes in synovial tissues and validated their roles on apoptosis and inflammation of OA synovial cells. Weighted correlation network analysis (WGCNA) was employed to explore OA-related co-expression modules in the GSE55235 and GSE55457 datasets. Then, this study screened OA-specific genes. After validation of these genes in the GSE12021 and GSE32317 datasets, HTR2B and SLC5A3 were obtained. Their expression was detected in human OA and healthy synovial tissues by RT-qPCR and western blot. OA rat models were constructed by anterior cruciate ligament transection (ACLT) operation. In OA synovial cells, HTR2B and SLC5A3 proteins were examined via western blot. After transfection with sh-HTR2B or sh-SLC5A3, apoptosis and inflammation of OA synovial cells were investigated by flow cytometry and western blot. A total of 17 OA-specific DEGs were identified, which were significantly enriched in inflammation pathways. Among them, HTR2B and SLC5A3 were highly expressed in end-than early-stage OA. Their up-regulation was validated in human OA synovial tissues and ACLT-induced OA synovial cells. Knockdown of HTR2B and SLC5A3 restrained apoptosis and increased TGF-β and IL-4 expression as well as reduced TNF-α and IL-1β expression in OA synovial cells. Collectively, this study identified two OA-specific markers HTR2B and SLC5A3 and their knockdown ameliorated apoptosis and inflammation of OA synovial cells.
骨关节炎（OA）是一种异质性的与年龄相关的疾病，由于其发病机制的复杂调控网络，很难治愈。本研究探索了滑膜组织中OA特异性基因，并验证了它们在OA滑膜细胞凋亡和炎症中的作用。采用加权基因共表达网络分析（WGCNA）在GSE55235和GSE55457数据集中探索与OA相关的共表达模块。然后，本研究筛选出OA特异性基因。在GSE12021和GSE32317数据集中对这些基因进行验证后，获得了HTR2B和SLC5A3。通过RT-qPCR和蛋白质印迹法检测它们在人OA和健康滑膜组织中的表达。通过前交叉韧带横断（ACLT）手术构建OA大鼠模型。在OA滑膜细胞中，通过蛋白质印迹法检测HTR2B和SLC5A3蛋白。用sh-HTR2B或sh-SLC5A3转染后，通过流式细胞术和蛋白质印迹法研究OA滑膜细胞的凋亡和炎症。共鉴定出17个OA特异性差异表达基因（DEGs），它们在炎症途径中显著富集。其中，HTR2B和SLC5A3在OA晚期比早期高表达。它们的上调在人OA滑膜组织和ACLT诱导的OA滑膜细胞中得到验证。敲低HTR2B和SLC5A3可抑制OA滑膜细胞凋亡，增加TGF-β和IL-4表达，并降低TNF-α和IL-1β表达。总体而言，本研究鉴定出两个OA特异性标志物HTR2B和SLC5A3，敲低它们可改善OA滑膜细胞的凋亡和炎症。
