Liao Rongdong, Feng Zhuoxi, Li Wei, Liu Rubing, Xu Xinrou, Yao Shun, Tian Jing
Department of Orthopedics, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510280, P.R. China.
The Second Clinical Medical School, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510280, P.R. China.
Exp Ther Med. 2021 Jun;21(6):640. doi: 10.3892/etm.2021.10072. Epub 2021 Apr 16.
Interleukin-1 (IL-1) is a pro-inflammatory cytokine which induces bone destruction in various diseases, such as osteoporosis and rheumatoid arthritis. RAW264.7 cells are frequently used in studies as osteoclast precursors, however it remains unclear whether IL-1 can induce osteoclast differentiation from RAW264.7 cells without the stimulation of receptor activator of nuclear factor-κB ligand (RANKL). Hence, the present study aimed to investigate the effects of IL-1 on the formation of osteoclasts from RAW264.7 cells. The cell viability was determined via the Cell Counting Kit-8 (CCK-8) assay. Protein and gene expression were measured by western blotting and reverse transcription-quantitative PCR, respectively. Tartrate-resistant acid phosphatase (TRAP) staining and the resorption pit assay were performed to determine the formation and activity of osteoclasts. A significantly increased quantity of osteoclasts were found in the IL-1 group compared with the control group, and also in the RANKL+IL-1 group compared with the RANKL group. In addition IL-1 significantly increased both the protein and mRNA expression of specific genes associated with osteoclastogenesis, including nuclear factor of activated T cells cytoplasmic 1, matrix metalloprotein-9, cathepsin K and TRAP. The findings of the present study suggested that IL-1 can induce osteoclast differentiation and upregulate the quantity of osteoclasts differentiated from RAW264.7 cells. These results may lay a foundation for further study of diseases involving inflammation-associated bone loss. The combined blockade of IL-1 and RANKL may be effective for the prevention of inflammatory bone loss.
白细胞介素-1(IL-1)是一种促炎细胞因子,可在多种疾病(如骨质疏松症和类风湿性关节炎)中诱导骨质破坏。RAW264.7细胞常作为破骨细胞前体用于研究,然而,尚不清楚IL-1在无核因子κB受体激活剂配体(RANKL)刺激的情况下能否诱导RAW264.7细胞分化为破骨细胞。因此,本研究旨在探讨IL-1对RAW264.7细胞形成破骨细胞的影响。通过细胞计数试剂盒-8(CCK-8)检测法测定细胞活力。分别通过蛋白质印迹法和逆转录定量PCR检测蛋白质和基因表达。进行抗酒石酸酸性磷酸酶(TRAP)染色和吸收陷窝试验以确定破骨细胞的形成和活性。与对照组相比,IL-1组中破骨细胞数量显著增加,与RANKL组相比,RANKL+IL-1组中破骨细胞数量也显著增加。此外,IL-1显著增加了与破骨细胞生成相关的特定基因的蛋白质和mRNA表达,包括活化T细胞核因子细胞质1、基质金属蛋白酶-9、组织蛋白酶K和TRAP。本研究结果表明,IL-1可诱导破骨细胞分化并上调RAW264.7细胞分化产生的破骨细胞数量。这些结果可能为进一步研究涉及炎症相关骨质流失的疾病奠定基础。联合阻断IL-1和RANKL可能对预防炎症性骨质流失有效。