Dalian Medical University, Dalian, Liaoning, PR China.
Department of Orthopedic Trauma, East Hospital, Tongji University School of Medicine, 150 Jimo Rd, 200120 Shanghai, PR China.
Fitoterapia. 2020 Apr;142:104392. doi: 10.1016/j.fitote.2019.104392. Epub 2020 Feb 27.
Previous studies have shown that mitomycin C (MMC) can prevent scar adhesion after joint surgery, but the specific mechanism underlying this effect remains unclear. The purpose of this study was to explore the specific mechanism by which MMC promotes fibroblast apoptosis and prevents joint adhesion. The effect of MMC on fibroblasts was assessed using cell counting kit-8 (CCK-8) assays, western blotting, and TUNEL staining. We used qRT-PCR to measure the expression of miR-21 in fibroblasts treated with MMC. Luciferase activity assays were used to determine the relationships between miR-21 and Programmed cell death 4 (PDCD4). The effects of miR-21 and PDCD4 on fibroblast apoptosis were assessed using flow cytometry and western blotting. HE staining was used to determine the role of miR-21 in scar tissue formation in a model of joint adhesion. The results showed that MMC induced apoptosis of fibroblasts and decreased the expression of miR-21. Moreover, miR-21 down-regulation also induced apoptosis of fibroblasts. PDCD4 was confirmed to be a direct target of miR-21 by luciferase activity assay. The results from the animal model indicated that miR-21 attenuated the effect of MMC on reducing the number of fibroblasts. Our study shows that MMC can induce fibroblast apoptosis and prevent joint adhesion by regulating the expression of miR-21 and its target PDCD4.
先前的研究表明丝裂霉素 C（MMC）可预防关节手术后的疤痕粘连，但这种作用的具体机制尚不清楚。本研究旨在探讨 MMC 促进纤维母细胞凋亡和防止关节粘连的具体机制。使用细胞计数试剂盒（CCK-8）测定、western blot 和 TUNEL 染色评估 MMC 对成纤维细胞的影响。用 qRT-PCR 测定 MMC 处理的成纤维细胞中 miR-21 的表达。用荧光素酶活性测定确定 miR-21 与程序性细胞死亡蛋白 4（PDCD4）之间的关系。用流式细胞术和 western blot 评估 miR-21 和 PDCD4 对纤维母细胞凋亡的影响。HE 染色用于确定 miR-21 在关节粘连模型中对疤痕组织形成的作用。结果表明，MMC 诱导成纤维细胞凋亡并降低 miR-21 的表达。此外，miR-21 的下调也诱导成纤维细胞凋亡。荧光素酶活性测定证实 PDCD4 是 miR-21 的直接靶标。动物模型的结果表明，miR-21 减弱了 MMC 减少成纤维细胞数量的作用。我们的研究表明，MMC 通过调节 miR-21 及其靶基因 PDCD4 的表达诱导纤维母细胞凋亡并防止关节粘连。
