Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
J Cell Physiol. 2019 Dec;234(12):21817-21824. doi: 10.1002/jcp.27316. Epub 2018 Nov 23.
To explore the effectiveness of the insulin-like growth factor 1 receptor (IGF-1R)/PI3K/AKT pathway in promoting the synthesis of the auricular chondrocyte extracellular matrix (ECM) using concentrated growth factor (CGF).
Chondrocytes isolated from the remnant auricular cartilage of microtia patients were randomly divided into different experimental and control groups, then stimulated with a reagent. IGF-1 released by CGF was quantified by enzyme-linked immunosorbent assay. Glycosaminoglycan (GAG), proteoglycan, and type II collagen (COLAII) were examined by histological and immunohistological analysis. Expression levels of IGF-1R, pIGF-1R, PI3K, pPI3K, AKT, pAKT, COLAII, and Aggrecan were detected by western blot analysis technique and gene expression was tested by real-time polymerase chain reaction.
CGF significantly stimulated the synthesis of COLAII and Aggrecan and increased the phosphorylation levels of IGF-1R, PI3K, and AKT. Small interfering RNA IGF-1 blocked ECM synthesis, COLAII and Aggrecan gene expression, and IGF-1R/PI3K/AKT activation. Inhibitor AG1024 and LY294002 significantly inhibited ECM synthesis and the phosphorylation of IGF-1R, PI3K, and AKT.
CGF-released IGF-1 stimulates the synthesis of the auricular chondrocyte ECM via the IGF-1R/PI3K/AKT signaling pathway.
探讨浓缩生长因子（CGF）中胰岛素样生长因子 1 受体（IGF-1R）/PI3K/AKT 通路在促进耳软骨细胞细胞外基质（ECM）合成中的作用。
将取自小耳畸形患者残余耳软骨的软骨细胞随机分为不同实验组和对照组，然后用试剂刺激细胞。通过酶联免疫吸附试验（ELISA）定量检测 CGF 释放的 IGF-1。通过组织学和免疫组织化学分析检测糖胺聚糖（GAG）、蛋白聚糖和 II 型胶原（COLAII）。通过 Western blot 分析技术检测 IGF-1R、pIGF-1R、PI3K、pPI3K、AKT、pAKT、COLAII 和 Aggrecan 的表达水平，并通过实时聚合酶链反应（PCR）检测基因表达。
CGF 可显著刺激 COLAII 和 Aggrecan 的合成，并增加 IGF-1R、PI3K 和 AKT 的磷酸化水平。IGF-1 小干扰 RNA（siRNA）阻断 ECM 合成、COLAII 和 Aggrecan 基因表达以及 IGF-1R/PI3K/AKT 激活。IGF-1R 抑制剂 AG1024 和 PI3K 抑制剂 LY294002 显著抑制 ECM 合成和 IGF-1R、PI3K 和 AKT 的磷酸化。
CGF 释放的 IGF-1 通过 IGF-1R/PI3K/AKT 信号通路刺激耳软骨细胞 ECM 的合成。
