骨关节炎软骨细胞对白细胞介素18的炎症应答

Inflammatory responses of human osteoarthritic chondrocytes induced by interleukin 18

目的探讨IL-18对骨关节炎软骨细胞的作用。方法取骨关节炎患者的膝关节软骨,进行原代细胞培养。不同浓度的IL-18(0,50,150,300 ng/m L)刺激软骨细胞,RT-PCR检测TNFα和COX-2 m RNA的表达,ELISA检测TNFα、PGE2的水平。应用IL-1受体阻断剂(IL-1Ra)+IL-18干预软骨细胞,检测软骨细胞COX-2的表达量和培养液中PGE2的水平。提取软骨细胞RNA,测定IL-18受体的表达。结果对于COX-2和TNFα,300 ng/m L组和150 ng/m L组m RNA的表达量显著高于对照组(P<0.01,P<0.05)。50、300 ng/m L组的PGE2水平显著高于对照组(P<0.05)。150、300 ng/m L组的TNFα蛋白的浓度显著高于对照组(P<0.05),IL-1Ra组的PGE2浓度高于对照组(P<0.01),但是低于IL-18组(P<0.01)。软骨细胞能够检测到IL-18受体的表达。结论 IL-18诱导软骨细胞产生炎症应答,这种作用和IL-1β有关但不完全依赖IL-1β。

Objective To explore the role of interleukin 18（IL-18） on the osteoarthritis（OA） patients＇ chondrocytes.Methods Knee cartilage samples were obtained from osteoarthritic patients,and the primary cells were cultured.After stimulated by IL-18（0,50,150,300 ng/m L）,chondrocytes were collect to obtain the total RNA,and the supernatant were extracted,too.The m RNA＇s expression of COX-2 and TNFα were detected by quantitative RT-PCR and the level of PGE2 and TNFα were investigated using ELISA.Besides IL-18,IL-1 receptor inhibitor（IL-1Ra） was also added into the medium of cell culture.COX-2 m RNA and PGE2 were respectively determination.IL-18 receptors in chondrocytes were detected by RT-PCR.Results m RNA expression of COX-2 and TNFα in 150 and 300 ng/m Lwere both significantly higher than that in control group（P0.05）.The level of in 50 and 300 ng/m L group were higher than that in control group（P0.01）.While the concentration of TNFα in150 and 300 ng/m L groups appeared more than that in control group significantly（P0.01）.And the density of PGE2 in IL-Ra group was significantly more than that in control group,but less than that of IL-18 group.IL-18 R can be detected on chondrocytes.Conclusion IL-18 induces inflammatory responses in osteoarthritic chondrocytes and that this effect was related with,although not entirely dependent on,IL-1β.