Geng Qin, Zhang Hongju, Cui Yanhui, Wei Qiaofeng, Wang Shujun
Department of Rheumatology, Shandong Zibo Central Hospital Zibo 255036, Shandong, China.
Am J Transl Res. 2021 Mar 15;13(3):979-987. eCollection 2021.
Osteoarthritis (OA) is a disease commonly diagnosed in the elderly population. It is reported that the reduction of extracellular matrix and infiltrated inflammation are two main factors responsible for the pathogenesis of OA. This investigation aims to explore the potential protective effects of Febuxostat against IL-18-induced insults in chondrocytes, as well as the possible mechanism.
The viability of chondrocytes was evaluated using the MTT assay. QRT-PCR and ELISA were used to measure the expressions and concentrations of IL-6, TNF-α, and CCL5, respectively. The accumulation of glycosaminoglycans (GAGs) was measured using Alcian blue assay. The chondrocytes were transfected with siRNA against Sox-9 in order to establish the Sox-9 knock-down chondrocytes. The expressions of were measured using qRT-PCR. The production of NO was measured using Diaminofluorescein-FM diacetate (DAF-FM DA) staining.
The up-regulated expressions of IL-6, TNF-α, CCL5, iNOS, and NO stimulated by IL-18 were down-regulated by the introduction of Febuxostat. The expressions of were significantly reduced by IL-18 but greatly promoted by Febuxostat. The increased gene expressions of and induced by Febuxostat were abolished by knocking down Sox-9 in the chondrocytes.
Febuxostat might mitigate IL-18-induced inflammatory response and reduction of the extracellular matrix gene mediated by Sox-9.
骨关节炎（OA）是一种在老年人群中常见的疾病。据报道，细胞外基质减少和炎症浸润是导致OA发病的两个主要因素。本研究旨在探讨非布索坦对白细胞介素-18（IL-18）诱导的软骨细胞损伤的潜在保护作用及其可能机制。
采用MTT法评估软骨细胞的活力。分别用实时定量聚合酶链反应（QRT-PCR）和酶联免疫吸附测定（ELISA）检测白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和趋化因子配体5（CCL5）的表达和浓度。采用阿尔辛蓝法检测糖胺聚糖（GAGs）的积聚。用针对性别决定区Y框蛋白9（Sox-9）的小干扰RNA（siRNA）转染软骨细胞，以建立Sox-9基因敲低的软骨细胞。用qRT-PCR检测相关基因的表达。用二氨基荧光素-FM二乙酸酯（DAF-FM DA）染色法检测一氧化氮（NO）的产生。
非布索坦可下调IL-18刺激引起的IL-6、TNF-α、CCL5、诱导型一氧化氮合酶（iNOS）和NO的上调表达。IL-18可显著降低相关基因的表达，但非布索坦可显著促进其表达。在软骨细胞中敲低Sox-9可消除非布索坦诱导的相关基因表达增加。
非布索坦可能减轻IL-18诱导的炎症反应以及Sox-9介导的细胞外基质基因减少。
