罗氟司特可预防淋巴毒素 α（TNF-β）诱导的软骨细胞中炎症激活和 2 型胶原的降解。
Roflumilast prevents lymphotoxin α (TNF-β)-induced inflammation activation and degradation of type 2 collagen in chondrocytes.
机构信息
Department of Central Laboratory, China-Japan Union Hospital of Jilin University, No. 126 Xiantai Street, Changchun, 130033, China.
Department of Radiotherapy, Tonghua Central Hospital, Tonghua, 134001, China.
出版信息
Inflamm Res. 2020 Dec;69(12):1191-1199. doi: 10.1007/s00011-020-01404-3. Epub 2020 Sep 29.
BACKGROUND AND PURPOSE
Osteoarthritis (OA) is a chronic disease accompanied by severe inflammation. The inflammation activation in the chondrocytes and the degradation of the extracellular matrix were reported to be involved in the progress of OA. Roflumilast is a selective PDE4 inhibitor used for treating chronic obstructive pulmonary disease (COPD) and exerts significant anti-inflammation effects. The present study aims to investigate the effects of Roflumilast on tumor necrosis factor-β (TNF-β)-induced inflammation activation and degradation of type 2 collagen in chondrocytes.
METHODS
TNF-β was used to establish the in-vitro inflammation model on ATDC5 chondrocytes. Quantitative real-time polymerase chain reaction (QRT-PCR) and western blot were used to determine the expression level of tumor necrosis factor receptor 2 (TNFR2), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), matrix metalloproteinase 3 (MMP-3), matrix metalloproteinase 13 (MMP-13), type 2 collagen and nuclear factor kappa B (NF-κB) p65. The release of prostaglandin E2 (PGE), MMP-3, and MMP-13 were evaluated by ELISA. The production of NO was determined by DAF-FM DA staining and the function of the NF-κB promoter was evaluated by Luciferase activity assay.
RESULTS
TNFR2 and COX-2 were upregulated and the release of PGE was promoted by TNF-β stimulation, which were all inhibited by Roflumilast. Roflumilast suppressed the promoted iNOS expression and NO production induced by TNF-β. MMP-3 and MMP-13 were up-regulated, and type 2 collagen was down-regulated by TNF-β stimulation, which were all reversed by Roflumilast. Roflumilast inhibited the promoted releasing of Interleukin-8 (IL-8) and Interleukin-12 (IL-12), expression of up-regulated NF-κB, and activation of NF-κB transcriptional activity induced by TNF-β.
CONCLUSION
Roflumilast may prevent TNF-β-induced inflammation activation and degradation of type 2 collagen in chondrocytes.
背景与目的
骨关节炎（OA）是一种伴有严重炎症的慢性疾病。据报道，软骨细胞中的炎症激活和细胞外基质的降解参与了 OA 的进展。罗氟司特是一种用于治疗慢性阻塞性肺疾病（COPD）的选择性 PDE4 抑制剂，具有显著的抗炎作用。本研究旨在探讨罗氟司特对肿瘤坏死因子-β（TNF-β）诱导的软骨细胞炎症激活和 II 型胶原降解的影响。
方法
采用 TNF-β 建立 ATDC5 软骨细胞体外炎症模型。实时定量聚合酶链反应（QRT-PCR）和 Western blot 用于检测肿瘤坏死因子受体 2（TNFR2）、环氧化酶-2（COX-2）、诱导型一氧化氮合酶（iNOS）、基质金属蛋白酶 3（MMP-3）、基质金属蛋白酶 13（MMP-13）、II 型胶原和核因子 kappa B（NF-κB）p65 的表达水平。ELISA 法检测前列腺素 E2（PGE）、MMP-3 和 MMP-13 的释放。通过 DAF-FM DA 染色测定 NO 的产生，通过荧光素酶活性测定评估 NF-κB 启动子的功能。
结果
TNF-β 刺激可上调 TNFR2 和 COX-2，促进 PGE 的释放，这些作用均被罗氟司特抑制。罗氟司特抑制 TNF-β 诱导的 iNOS 表达和 NO 产生的增加。TNF-β 刺激可上调 MMP-3 和 MMP-13，下调 II 型胶原，这些作用均被罗氟司特逆转。罗氟司特抑制 TNF-β 诱导的白细胞介素-8（IL-8）和白细胞介素-12（IL-12）释放增加、NF-κB 表达上调以及 NF-κB 转录活性的激活。
结论
罗氟司特可能预防 TNF-β 诱导的软骨细胞炎症激活和 II 型胶原降解。
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