Protective effects of cilengitide on inflammation in chondrocytes under excessive mechanical stress
Protective effects of cilengitide on inflammation in chondrocytes under excessive mechanical stress
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DOI：
10.1002/cbin.11293
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发表时间：
2020-01-06
影响因子：
3.9
通讯作者：
Tanimoto, Kotaro
中科院分区：
文献类型：
--
作者：
Hirose, Naoto;Okamoto, Yuki;Tanimoto, Kotaro
软骨细胞不断地接受外界刺激，从而调节重塑。适宜的机械应力水平对于维持软骨细胞内环境的稳定是必不可少的，但过高的机械应力会诱导炎性细胞因子和蛋白酶，如基质金属蛋白酶(MMPs)。因此，过大的机械应力被认为是导致软骨破坏导致骨关节炎的主要原因之一。整合素是众所周知的细胞黏附分子，是细胞外基质(ECM)的受体，被认为作为机械感受器在物理和化学上控制细胞内的信号通路。然而，很少有研究关注整合素在过度机械应力引起的炎症中的作用和功能。在这项研究中，我们使用整合素受体拮抗剂(Cilengiide)，研究了在机械负荷下，软骨细胞中整合素αVβ3和αVβ5与炎症因子表达的关系。Cilengiide抑制过度机械应激诱导的白介素1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、基质金属蛋白酶-3(MMP3)和MMP13的基因表达。此外，加入西伦吉肽还可抑制IL-1-β和基质金属蛋白酶-13的蛋白表达。接下来，我们通过Western blotting研究了细胞内信号通路在应激诱导的软骨细胞整合素信号转导中的作用。过度机械应力可使p-FAK、p-ERK、p-JNK和p-p38水平升高，而cilengitide可抑制这种升高。综上所述，本研究揭示了过度的机械应力可激活软骨细胞表面的整合素αVβ3和αVβ5，从而通过FAK和MAPKs的磷酸化上调IL-1β、肿瘤坏死因子-α、基质金属蛋白酶-3和基质金属蛋白酶-13的表达，从而诱导炎症反应。
Chondrocytes constantly receive external stimuli, which regulates remodeling. An optimal level of mechanical stress is essential for maintaining chondrocyte homeostasis, however, excessive mechanical stress induces inflammatory cytokines and protease, such as matrix metalloproteinases (MMPs). Therefore, excessive mechanical stress is considered to be one of the main causes to cartilage destruction leading to osteoarthritis (OA). Integrins are well-known as cell adhesion molecules and act as receptors for extracellular matrix (ECM), and are believed to control intracellular signaling pathways both physically and chemically as a mechanoreceptor. However, few studies have focused on the roles and functions of integrins in inflammation caused by excessive mechanical stress. In this study, we examined the relationship between integrins (alpha V beta 3 and alpha V beta 5) and the expression of inflammatory factors under mechanical loading in chondrocytes by using an integrin receptor antagonist (cilengitide). Cilengitide suppressed the gene expression of interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), matrix metalloproteinase-3 (MMP-3), and MMP-13 induced by excessive mechanical stress. In addition, the protein expression of IL1-beta and MMP-13 was also inhibited by the addition of cilengitide. Next, we investigated the involvement of intracellular signaling pathways in stress-induced integrin signaling in chondrocytes by using western blotting. The levels of p-FAK, p-ERK, p-JNK, and p-p38 were enhanced by excessive mechanical stress and the enhancement was suppressed by treatment with cilengitide. In conclusion, this study revealed that excessive mechanical stress may activate integrins alpha V beta 3 and alpha V beta 5 on the surface of chondrocytes and thereby induce an inflammatory reaction by upregulating the expression of IL-1 beta, TNF-alpha, MMP-3, and MMP-13 through phosphorylation of FAK and MAPKs.