Hyaluronan suppresses mechanical stress-induced expression of catabolic enzymes by human chondrocytes via inhibition of IL-1β production and subsequent NF-κB activation
Hyaluronan suppresses mechanical stress-induced expression of catabolic enzymes by human chondrocytes via inhibition of IL-1β production and subsequent NF-κB activation
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DOI：
10.1007/s00011-015-0804-2
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发表时间：
2015-04-01
影响因子：
6.7
通讯作者：
Ozaki, Toshifumi
中科院分区：
文献类型：
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作者：
Ozawa, Masatsugu;Nishida, Keiichiro;Ozaki, Toshifumi
为研究透明质酸(HA)对机械应激诱导的人关节软骨细胞去整合素和基质金属蛋白酶(MMP13)表达的抑制作用，将人关节软骨细胞置于37℃、37℃、0.5 Hz、10%拉伸条件下，分别加入或不加1.0 mg/mLHA(2700 KDa)预孵育12h，观察透明质酸(HA)对人关节软骨细胞去整合素和金属蛋白酶表达的影响。用实时定量聚合酶链式反应和免疫细胞化学方法检测ADAMTS-4、-5和基质金属蛋白酶-13的表达。采用酶联免疫吸附试验(ELISA)检测培养上清液中IL-1β的浓度。免疫细胞化学检测核转录因子2(RUNX-2)和核因子-kappaB(NF-kappa B)的核转位，Western blotting检测核转录因子-2(RUNX-2)和核转录因子-kappaB(NFkappa B)的磷酸化。HA通过抑制IL-1β的分泌和核转录因子kappaB的活化抑制ADAMTS-4、-5和MMP13的表达。然而，HA不能抑制CTS诱导的RUNX-2的表达以及随后的ADAMTS-5和MMP-13的表达。结果表明，HA通过抑制NF-kappa B-IL-1β途径显著抑制机械应激诱导的分解蛋白的表达，但不抑制机械应激诱导的RUNX-2信号转导。
To investigate the inhibitory effect of hyaluronan (HA) on mechanical stress- induced expression of a disintegrin and metalloproteinase with thrombospondin type1 motifs (ADAMTS)-4, -5 and matrix metalloproteinase (MMP)-13 by human chondrocytes.Normal human articular chondrocytes were pre-incubated with or without 1.0 mg/mL HA (2700 kDa) for 12 h at 37 A degrees C in stretch chambers, then they were exposed to uni-axial cyclic tensile strain (CTS, 0.5 Hz, 10 % elongation). The expression of ADAMTS-4, -5, and MMP-13 were analyzed by real-time polymerase chain reaction and Immunocytochemistry. The concentration of IL-1 beta in the supernatant was measured using enzyme-linked immunosorbent assay (ELISA). The nuclear translocation of runt-related transcription factor 2 (RUNX-2) and nuclear factor-kappa B (NF-kappa B) was examined by ELISA and immunocytochemistry, and phosphorylation of NF-kappa B was examined by western blotting.HA inhibited mRNA expression of ADAMTS-4, -5, and MMP13 after 24 h CTS via inhibition of IL-1 beta secretion and NF-kappa B activation. However, HA failed to inhibit CTS-induced RUNX-2 expression and subsequent expression of ADAMTS-5 and MMP-13 1 h after CTS.Our results demonstrated that HA significantly suppressed mechanical stress-induced expression of catabolic proteases by inhibition of the NF-kappa B-IL-1 beta pathway, but did not suppress mechanical stress-induced RUNX-2 signaling.