Yang Y, Gao S-G, Zhang F-J, Luo W, Xue J-X, Lei G-H
Department of Orthopedics, XiangYa Hospital, Central South University, Changsha, China.
Eur Rev Med Pharmacol Sci. 2014;18(23):3580-6.
Osteoarthritis (OA) is a chronic musculoskeletal disease characterized by progressive destruction of articular cartilage, OA lead to chronic pain and functional restrictions in affected joints. The present study was to investigate the role of osteopontin (OPN) in the athogenesis of OA through studying the effect of OPN on expression of IL-6 and IL-8 inflammatory factors in human OA chondrocytes.
One-step type II collagenase digestive method was used to isolate OA chondrocytes from sectional cartilage specimens of 16 primary knee OA patients received total knee replacement surgery. Synchronized first-generation chondrocytes were then treated with OPN (100 ng/ml or 1 µg/mL). The changes in cell morphology of OA chondrocytes were analyzed before and after treated with OPN; and the expression levels of IL-6 and IL-8 were evaluated by real-time q-PCR.
Chondrocytes were successfully isolated from human OA knee cartilage, and the viability of isolated chondrocytes was 92.11±3.13%. Adherent chondrocytes formed clusters of irregular polygonal shape with intercellular pseudopodia extension. After OPN treatment, cells became fusiform or irregularly shaped, and the number of intercellular pseudopodia decreased significantly. The mRNA expression of IL-6 increased to 1.83 times at 0.1 µg/ml of OPN and 3.1 times at the dose of 1 µg/ml; the expression of IL-8 increased to 1.57 and 3.27 times at the dose of 0.1 µg/ml and 1 µg/ml respectively.
OPN could up-regulate expression of IL-6 and IL-8 cytokines in human OA chondrocytes, and the expression increased with the increasing concentration of OPN, which might be one of the potential mechanisms of OPN in the development of OA.
骨关节炎(OA)是一种慢性肌肉骨骼疾病,其特征是关节软骨进行性破坏,OA会导致受影响关节出现慢性疼痛和功能受限。本研究旨在通过研究骨桥蛋白(OPN)对人OA软骨细胞中白细胞介素-6(IL-6)和白细胞介素-8(IL-8)炎症因子表达的影响,探讨OPN在OA发病机制中的作用。
采用一步法II型胶原酶消化法,从16例行全膝关节置换手术的原发性膝OA患者的关节软骨标本中分离OA软骨细胞。同步化的第一代软骨细胞随后用OPN(100 ng/ml或1 μg/mL)处理。分析OPN处理前后OA软骨细胞的细胞形态变化;并通过实时定量聚合酶链反应评估IL-6和IL-8的表达水平。
成功从人OA膝关节软骨中分离出软骨细胞,分离出的软骨细胞活力为92.11±3.13%。贴壁的软骨细胞形成不规则多边形簇,细胞间有伪足延伸。OPN处理后,细胞变为梭形或不规则形,细胞间伪足数量明显减少。在0.1 μg/ml的OPN作用下,IL-6的mRNA表达增加到1.83倍,在1 μg/ml剂量下增加到3.1倍;在0.1 μg/ml和1 μg/ml剂量下,IL-8的表达分别增加到1.57倍和3.27倍。
OPN可上调人OA软骨细胞中IL-6和IL-8细胞因子的表达,且表达随OPN浓度的增加而增加,这可能是OPN在OA发展中的潜在机制之一。