Inflammatory responses of human osteoarthritic chondrocytes induced by interleukin 18
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摘要:
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/mL）刺激软骨细胞，RT-PCR检测TNFα和COX-2 mRNA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/mL组和150 ng/mL组mRNA的表达量显著高于对照组（P<0.01，P<0.05）。50、300 ng/mL组的PGE2水平显著高于对照组（P<0.05）。150、300 ng/mL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。 Abstract: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/mL), chondrocytes were collect to obtain the total RNA, and the supernatant were extracted, too. The mRNA’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 mRNA and PGE2 were respectively determination. IL-18 receptors in chondrocytes were detected by RT-PCR. Results mRNA expression of COX-2 and TNFα in 150 and 300 ng/mLwere both significantly higher than that in control group (P<0.05). The level of in 50 and 300 ng/mL group were higher than that in control group (P<0.01). While the concentration of TNFα in 150 and 300 ng/mL groups appeared more than that in control group significantly (P<0.01). 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-18R can be detected on chondrocytes. Conclusions IL-18 induces inflammatory responses in osteoarthritic chondrocytes and that this effect was related with, although not entirely dependent on, IL-1β. -
Key words:
- osteoarthritis /
- interleukin 18 /
- chondrocytes /
- degradation
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图 1 软骨细胞的形态图
A: 显示细胞接种24 h, 部分细胞贴壁，未贴壁细胞呈球形; B: 显示典型软骨细胞呈多边形; C: 软骨细胞鉴定，棕色区域为Ⅱ型胶原阳性染色, 分布于胞浆和细胞间; D: 对照组,无阳性染色. 标记线为100 μm.
表 1 IL-18 干预后PGE2、TNF-α的水平(x±s, pg/mL)
Group PGE2 (n=8) TNF-α (n=8) 0 ng/mL 340.47±22.03 20.48±7.36 50 ng/mL 410.03±22.97* 25.32±9.55 150 ng/mL 460.38±112.86* 52.45±10.93# 300 ng/mL 518.28±38.80* 103.80±22.29# *P<0.01 vs PGE2 in 0 ng/mL group; #P<0.01 vs TNF-α in 0 ng/mL group. 
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