Department of Pain Management, Linyi People's Hospital, Linyi, Shandong, China.
Department of Blood Collection, Linyi Central Blood Station, Linyi, Shandong, China.
J Cell Physiol. 2019 Aug;234(8):12771-12785. doi: 10.1002/jcp.27897. Epub 2018 Dec 12.
This study aims to test the effect of naproxen treatment and the biological target of naproxen for treating osteoarthritis (OA).
Differentially expressed genes (DEGs) in OA synovial tissues and normal counterparts were analyzed by messenger RNA microarray analysis. R package (weighted gene coexpression network analysis) was used to divide DEGs into several modules and determine the hub genes in each module. The expression level of prostaglandin-endoperoxide synthase 1 ( PTGS1) in OA synovial cells and tissues was verified by a quantitative real-time polymerase chain reaction and western blot. Transwell assay evaluated the numbers of cell migration and invasion. Furthermore, Safranin O and fast green staining and hematoxylin and eosin staining were performed on joints from anterior cruciate ligament transection mice.
Microarray analysis determined PTGS1 was the hub gene in the black module, which was overexpressed in OA synovial cells and tissues compared with normal synovial cells. OA synovial cells transfected with sh-PTGS1 showed downregulation of PTGS1. After treatment with naproxen, the expression of PTGS1 sharply decreased in the OA group. The migration and invasion of OA synovial cells increased, whereas the cell apoptosis rate decreased when PTGS1 was overexpressed. However, the cell migration and invasion decreased, whereas cells apoptosis increased when it was treated with naproxen. Naproxen could also influence the expression level of six OA-related genes: LUBRICIN, matrix metalloproteinase 13 (MMP-13), cyclooxygenase-2 (COX-2), ACAN, COL2A1, and COL1A1.
We validated that naproxen could suppress the expression of PTGS1 in synovial cells. Moreover, naproxen could inhibit the migration/invasion ability of OA synoviocytes and promote the apoptosis rate OA synoviocytes.
本研究旨在测试萘普生治疗骨关节炎（OA）的疗效及其生物靶点。
采用信使 RNA 微阵列分析分析 OA 滑膜组织与正常对照组织中的差异表达基因（DEGs）。R 包（加权基因共表达网络分析）用于将 DEGs 分为几个模块，并确定每个模块中的枢纽基因。通过实时定量聚合酶链反应和 Western blot 验证前列腺素内过氧化物合酶 1（PTGS1）在 OA 滑膜细胞和组织中的表达水平。Transwell 测定细胞迁移和侵袭的数量。此外，对前交叉韧带切断小鼠的关节进行番红 O 和固绿染色以及苏木精和伊红染色。
微阵列分析确定 PTGS1 是黑色模块中的枢纽基因，其在 OA 滑膜细胞和组织中的表达高于正常滑膜细胞。与 sh-PTGS1 转染的 OA 滑膜细胞相比，PTGS1 表达下调。在用萘普生处理后，OA 组中 PTGS1 的表达急剧下降。OA 滑膜细胞中 PTGS1 的过表达会增加细胞的迁移和侵袭，而降低细胞凋亡率。但是，当用萘普生处理时，细胞迁移和侵袭减少，而细胞凋亡增加。萘普生还可以影响六个 OA 相关基因的表达水平：润滑素、基质金属蛋白酶 13（MMP-13）、环氧化酶 2（COX-2）、ACAN、COL2A1 和 COL1A1。
我们验证了萘普生可以抑制滑膜细胞中 PTGS1 的表达。此外，萘普生可以抑制 OA 滑膜细胞的迁移/侵袭能力并促进 OA 滑膜细胞的凋亡率。
