Overexpression of SOX9 alleviates the progression of human osteoarthritis in vitro and in vivo
Overexpression of SOX9 alleviates the progression of human osteoarthritis in vitro and in vivo
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DOI：
10.2147/dddt.s203974
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发表时间：
2019-01-01
影响因子：
4.8
通讯作者：
Li, Yanfeng
中科院分区：
文献类型：
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作者：
Ouyang, Yuanming;Wang, Wei;Li, Yanfeng
目的：最近的研究发现SOX9在骨关节炎(OA)的发病机制中起着关键作用。本研究旨在探讨SOX9在体内外调控骨性关节炎形成的机制。材料与方法：采用实时荧光定量聚合酶链式反应技术检测骨性关节炎患者及正常股骨骨折组织中SOX9的相对表达。在体外，采用IL-1β诱导的人软骨细胞炎症反应来评价SOX9的功能。利用重组SOX9慢病毒载体(Lenti-SOX9)上调SOX9在细胞中的表达。采用双抗体夹心法测定血清肿瘤坏死因子-α(TNF-α)浓度。免疫印迹法检测SOX9、基质金属蛋白酶-13(MMP13)、II型胶原、聚集素和Smad3的蛋白表达。CCK-8比色法检测细胞增殖，流式细胞仪检测细胞凋亡。结果：与正常人相比，骨性关节炎患者外周血中SOX9基因表达水平明显下调，而肿瘤坏死因子-α水平明显上调。此外，IL-1β还可降低软骨细胞SOX9、II型胶原和聚集素的表达，增加MMP13的表达。此外，Lenti-SOX9通过上调Smad3的表达，显著抑制IL-1β诱导的软骨细胞生长抑制和细胞凋亡。结论：上调SOX9可通过增加人软骨细胞Smad3水平抑制IL-1β诱导的炎症反应，对手术诱导的骨性关节炎小鼠具有体内治疗作用。因此，SOX9可能成为未来治疗骨性关节炎的潜在靶点。
Purpose: Recent findings have identified that SOX9 served as a key role during the pathogenesis of osteoarthritis (OA). This study aimed to investigate the mechanisms by which SOX9 regulated the formation of OA in vitro and in vivo.Materials and methods: The relative expressions of SOX9 in patients with OA and normal fracture of thighbone were analyzed by real-time-PCR. In vitro, IL-1 beta induced inflammatory response in human chondrocytes was used to evaluate the function of SOX9. The recombinant SOX9 lentivirus vector (Lenti-SOX9) was used to upregulate the expression of SOX9 in cells. ELISA was used to measure the concentration of tumornecrosis factor-alpha (TNF-alpha). The protein expressions of SOX9, matrixmetalloproteinase-13 (MMP13), Collagen II, Aggrecan and Smad3 were analyzed by Western blot. Cell proliferation and cell apoptosis were detected by CCK-8 assay and flow cytometry, respectively. In vivo, the effect of SOX9 on surgically induced OA mice was evaluated.Results: The gene level of SOX9 was remarkably downregulated in patients with OA compared with normal people, while the concentration of TNF-alpha was upregulated. In addition, IL-1 beta reduced the expressions of SOX9, Collagen II and Aggrecan and increased the level of MMP13 in chondrocytes. Moreover, Lenti-SOX9 notably inhibited IL-1 beta-induced growth inhibition and apoptosis in chondrocytes via increasing the expression of Smad3. Finally, Lenti-SOX9 markedly alleviated the symptoms of OA mice in vivo.Conclusion: Upregulation of SOX9 inhibited IL-1 beta-induced inflammatory response via increasing the level Smad3 in human chondrocytes and exhibited therapeutic effect on surgically induced OA mice in vivo. Therefore, SOX9 may serve as a potential target in the treatment of OA in the future.