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.
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。