Abstract：
BACKGROUND: Gelatinases, namely MMP2 and MMP9, are involved in the natural turnover of articular cartilage, as well as the loss of the cartilage matrix in osteoarthritis (OA). Studies have reported that fibroblast growth factor 8 (FGF8) promoted the degradation of cartilage in OA. In the present study, we predicted that FGF8 promoted chondrocyte expression and secretion of gelatinases by activating NF-κB p65 signaling.
METHODS: Primary chondrocytes from C57 mice were cultured with recombinant FGF8. RNA sequencing was employed to explore the gene expression changes of gelatinases. Gelatin zymography was used to determine the activation of gelatinases. Western blot was used to investigate the expression of the gelatinases and NF-κB p65 signaling pathways, and immunofluorescence staining and NF-κB inhibitor assays were performed to confirm the activation of NF-κB p65 signaling.
RESULTS: FGF8 could increase the expression and activity of gelatinases in primary chondrocytes. And FGF8-induced expression of gelatinases was regulated through activation of NF-κB signaling with acetylated p65 accumulating in the cell nucleus. We further found that the NF-κB inhibitor, BAY 11-7082, could suppress up-regulation of gelatinase induced by FGF8.
CONCLUSIONS: FGF8 enhanced the expression and activity of MMP2 and MMP9 in chondrocytes via NF-κB p65 signaling.
METHODS: Primary chondrocytes from C57 mice were cultured with recombinant FGF8. RNA sequencing was employed to explore the gene expression changes of gelatinases. Gelatin zymography was used to determine the activation of gelatinases. Western blot was used to investigate the expression of the gelatinases and NF-κB p65 signaling pathways, and immunofluorescence staining and NF-κB inhibitor assays were performed to confirm the activation of NF-κB p65 signaling.
RESULTS: FGF8 could increase the expression and activity of gelatinases in primary chondrocytes. And FGF8-induced expression of gelatinases was regulated through activation of NF-κB signaling with acetylated p65 accumulating in the cell nucleus. We further found that the NF-κB inhibitor, BAY 11-7082, could suppress up-regulation of gelatinase induced by FGF8.
CONCLUSIONS: FGF8 enhanced the expression and activity of MMP2 and MMP9 in chondrocytes via NF-κB p65 signaling.
摘要：
背景：明胶酶，即MMP2和MMP9，参与关节软骨的自然周转，以及骨关节炎（OA）中软骨基质的损失。有研究报道成纤维细胞生长因子8(FGF8)促进OA软骨的降解。在本研究中,我们预测FGF8通过激活NF-κBp65信号促进软骨细胞表达和明胶酶的分泌。
方法:用重组FGF8培养来自C57小鼠的原代软骨细胞。采用RNA测序技术探索明胶酶基因表达的变化。明胶酶谱用于确定明胶酶的活化。Westernblot检测明胶酶和NF-κBp65信号通路的表达,进行免疫荧光染色和NF-κB抑制剂测定以证实NF-κBp65信号的激活。
结果：FGF8可以增加原代软骨细胞中明胶酶的表达和活性。FGF8诱导的明胶酶的表达是通过激活NF-κB信号来调节的，乙酰化p65在细胞核中积累。我们进一步发现NF-κB抑制剂，BAY11-7082可以抑制FGF8诱导的明胶酶的上调。
结论：FGF8通过NF-κBp65信号增强软骨细胞中MMP2和MMP9的表达和活性。
方法:用重组FGF8培养来自C57小鼠的原代软骨细胞。采用RNA测序技术探索明胶酶基因表达的变化。明胶酶谱用于确定明胶酶的活化。Westernblot检测明胶酶和NF-κBp65信号通路的表达,进行免疫荧光染色和NF-κB抑制剂测定以证实NF-κBp65信号的激活。
结果：FGF8可以增加原代软骨细胞中明胶酶的表达和活性。FGF8诱导的明胶酶的表达是通过激活NF-κB信号来调节的，乙酰化p65在细胞核中积累。我们进一步发现NF-κB抑制剂，BAY11-7082可以抑制FGF8诱导的明胶酶的上调。
结论：FGF8通过NF-κBp65信号增强软骨细胞中MMP2和MMP9的表达和活性。
