Department of Orthopaedics, China-Japan Union Hospital of Jilin University, Changchun 130033, China.
Department of Orthopaedics, China-Japan Union Hospital of Jilin University, Changchun 130033, China.
Int Immunopharmacol. 2019 Jan;66:354-361. doi: 10.1016/j.intimp.2018.11.038. Epub 2018 Dec 4.
Osteoarthritis is an age-related disorder of bone-joint that causes pain and disability in middle and older people. This study aimed to investigate the potential effects of long non-coding RNA (lncRNA) THRIL on lipopolysaccharide (LPS)-induced osteoarthritis cell injury model (ATDC5 cell inflammatory injury), as well as the possible internal molecular mechanisms.
Cell viability and apoptosis were assessed using CCK-8 assay and Guava Nexin assay, respectively. Cell transfection was conducted to change the expression of THRIL and microRNA-125b (miR-125b) in ATDC5 cells. qRT-PCR was performed to detect the expression of THRIL, miR-125b and pro-inflammatory cytokines IL-6, TNF-α and monocyte chemotactic protein 1 (MCP-1) in ATDC5 cells. ELISA was used to measure the concentrations of IL-6, TNF-α and MCP-1 in culture supernatant of ATDC5 cells. Finally, the protein expression of key factors involved in cell apoptosis, inflammatory response, JAK1/STAT3 and NF-κB pathways were evaluated using western blotting.
LPS significantly induced ATDC5 cell inflammatory injury and up-regulated the expression of THRIL. Overexpression of THRIL aggravated the LPS-induced ATDC5 cell inflammatory injury. Suppression of THRIL had opposite effects. Moreover, THRIL negatively regulated the expression of miR-125b in ATDC5 cells. miR-125b participated in the effects of THRIL overexpression on LPS-induced ATDC5 cell inflammatory injury. Furthermore, overexpression of THRIL enhanced the LPS-induced JAK1/STAT3 and NF-κB pathways activation by down-regulating miR-125b.
THRIL exerted pro-inflammatory roles in LPS-induced osteoarthritis cell injury model. Overexpression of THRIL promoted LPS-induced ATDC5 cell inflammatory injury by down-regulating miR-125b and then activating JAK1/STAT3 and NF-κB pathways.
骨关节炎是一种与年龄相关的骨关节疾病，会导致中老年人疼痛和残疾。本研究旨在探讨长链非编码 RNA (lncRNA) THRIL 对脂多糖 (LPS) 诱导的骨关节炎细胞损伤模型 (ATDC5 细胞炎症损伤) 的潜在影响，以及可能的内在分子机制。
通过 CCK-8 测定法和 Guava Nexin 测定法分别评估细胞活力和细胞凋亡。通过细胞转染改变 ATDC5 细胞中 THRIL 和 microRNA-125b (miR-125b) 的表达。通过 qRT-PCR 检测 ATDC5 细胞中 THRIL、miR-125b 和促炎细胞因子 IL-6、TNF-α 和单核细胞趋化蛋白 1 (MCP-1) 的表达。ELISA 法检测 ATDC5 细胞培养上清液中 IL-6、TNF-α 和 MCP-1 的浓度。最后，通过 Western blot 法评估参与细胞凋亡、炎症反应、JAK1/STAT3 和 NF-κB 通路的关键因子的蛋白表达。
LPS 显著诱导 ATDC5 细胞炎症损伤，并上调 THRIL 的表达。THRIL 的过表达加重了 LPS 诱导的 ATDC5 细胞炎症损伤。THRIL 的抑制作用则相反。此外，THRIL 负调控 ATDC5 细胞中 miR-125b 的表达。miR-125b 参与了 THRIL 过表达对 LPS 诱导的 ATDC5 细胞炎症损伤的影响。此外，THRIL 的过表达通过下调 miR-125b 增强了 LPS 诱导的 JAK1/STAT3 和 NF-κB 通路的激活。
THRIL 在 LPS 诱导的骨关节炎细胞损伤模型中发挥促炎作用。THRIL 的过表达通过下调 miR-125b 促进 LPS 诱导的 ATDC5 细胞炎症损伤，进而激活 JAK1/STAT3 和 NF-κB 通路。
