Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Molecular Developmental Biology, School of Life Sciences, Southwest University, Chongqing, China.
Center for Joint Surgery, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, China.
Cell Prolif. 2019 Jan;52(1):e12532. doi: 10.1111/cpr.12532. Epub 2018 Oct 17.
To investigate the role of zyxin-involved actin regulation in expression level of vinculin focal adhesion and collagen production of chondrocyte and its possible underlying mechanism.
Chondrocytes obtained from rabbit articular cartilage were used in this study. The expression of zyxin, actin and vinculin, as well as the extracellular matrix (ECM) protein collagen type I, II and X (COL I, II and X) of chondrocytes were compared between zyxin-knockdown group and negative control group, and between transforming growth factor-β1 (TGF-β1) treatment group and non-treatment group, respectively.
Knockdown of zyxin increased the ratio of globular actin (G-actin) to filamentous actin (F-actin) of chondrocyte, which further inhibited expression of vinculin and chondrogenic marker COL II as well as hypertrophy marker COL X. On the other hand, chondrocytes treated with TGF-β1 showed an enhanced expression of F-actin, and a lower expression of zyxin compared to non-treatment group. In response to TGF-β1-induced actin polymerization, expression of vinculin and COL I was increased, while expression of COL II and aggrecan was decreased.
These results demonstrate supporting evidence that in chondrocytes the level of zyxin is closely associated with the state of actin polymerization. In particular, the change of zyxin and F-actin parallels with the change of COL II and vinculin, respectively, indicating a major role of zyxin-actin interaction in the synthesis of collagen ECM and the remodelling of cytoskeleton-ECM adhesion.
研究 zyxin 参与的肌动蛋白调节在软骨细胞黏附斑蛋白 vinculin 表达水平和胶原产生中的作用及其可能的潜在机制。
本研究采用兔关节软骨来源的软骨细胞。比较 zyxin 敲低组和阴性对照组以及转化生长因子-β1(TGF-β1)处理组和非处理组之间软骨细胞中 zyxin、肌动蛋白和黏附斑蛋白 vinculin 的表达,以及细胞外基质(ECM)蛋白Ⅰ型、Ⅱ型和 X 型胶原(COL I、II 和 X)的表达。
敲低 zyxin 增加了软骨细胞球状肌动蛋白(G-actin)与丝状肌动蛋白(F-actin)的比例,从而进一步抑制了黏附斑蛋白 vinculin 和软骨形成标志物 COL II 以及肥大标志物 COL X 的表达。另一方面,与非处理组相比,TGF-β1 处理的软骨细胞表现出 F-actin 的表达增强,而 zyxin 的表达降低。TGF-β1 诱导的肌动蛋白聚合反应导致黏附斑蛋白 vinculin 和 COL I 的表达增加,而 COL II 和聚集蛋白的表达减少。
这些结果为 zyxin 与肌动蛋白聚合状态密切相关提供了支持证据。特别是 zyxin 和 F-actin 的变化分别与 COL II 和 vinculin 的变化平行,表明 zyxin-actin 相互作用在胶原 ECM 合成和细胞骨架-ECM 黏附重塑中起着重要作用。