Gong Yong-Fang, Zhang Xiao-Ming, Liu Fei, Wang Zhen-Zhen, Deng Xue-Fei, Jiao Yi, Li Xiao-Jing, Huang Xue-Ying
Department of Anatomy, Anhui Medical University, 81 Meishan Road, Hefei 230032, China.
Department of Plastic Surgery, The First Affiliated Hospital, Anhui Medical University, 218 Jixi Road, Hefei 230022, China.
Eur J Pharmacol. 2016 Nov 15;791:647-654. doi: 10.1016/j.ejphar.2016.09.034. Epub 2016 Sep 28.
Hypertrophic scar (HS) is a pathological scar that particularly occurs after traumatic injuries, surgical procedures and burning. Abnormal activation of hypertrophic scar fibroblasts (HSFs) intensifies fibrosis during wound healing. Our previous studies demonstrated that recombinant human endostatin (rhEndostatin) prevented synovial thickening in adjuvant arthritis (AA) rats via inhibition of proliferation and enhancement of apoptosis in synovial fibroblasts. However, the effect of this protein on HSF proliferation is not known. This study investigated the inhibitory effect of rhEndostatin on the proliferation of cultured HSFs in a rabbit ear model. MTT assay and flow cytometric detection were performed to investigate HSF proliferation and cell cycle progression, respectively. The expression levels of p53, p21, cyclinD1, cyclin-dependent kinase 4 (CDK) and proliferating cell nuclear antigen (PCNA) in HSFs were detected using real-time PCR and Western blotting. Our data revealed that HSFs treated with rhEndostatin were significantly inhibited in a concentration-dependent manner with an IC value of 100mg/L. Also, rhEndostatin (100mg/L) primarily induced G/G and partially G/M cell cycle arrest of HSFs. There were significant decreases in the expression levels of p53, p27, CDK, cyclinD and PCNA in HSFs treated with rhEndostatin. In conclusion, rhEndostatin inhibited HSF proliferation via G/G and/or G/M phase arrest of the cell cycle, which was partially due to the down-regulation of cyclinD, CDK and PCNA. These findings suggest that rhEndostatin may reduce scar hypertrophy in vivo via inhibition of HSF proliferation and may be a novel agent for HS treatment.
肥厚性瘢痕（HS）是一种病理性瘢痕，尤其发生于创伤性损伤、外科手术和烧伤后。肥厚性瘢痕成纤维细胞（HSFs）的异常激活会加剧伤口愈合过程中的纤维化。我们之前的研究表明，重组人内皮抑素（rhEndostatin）通过抑制佐剂性关节炎（AA）大鼠滑膜成纤维细胞的增殖并增强其凋亡，从而预防滑膜增厚。然而，这种蛋白质对HSFs增殖的影响尚不清楚。本研究在兔耳模型中研究了rhEndostatin对培养的HSFs增殖的抑制作用。分别进行MTT法和流式细胞术检测以研究HSFs的增殖和细胞周期进程。使用实时PCR和蛋白质印迹法检测HSFs中p53、p21、细胞周期蛋白D1（cyclinD1）、细胞周期蛋白依赖性激酶4（CDK）和增殖细胞核抗原（PCNA）的表达水平。我们的数据显示，用rhEndostatin处理的HSFs受到显著抑制，呈浓度依赖性，IC值为100mg/L。此外，rhEndostatin（100mg/L）主要诱导HSFs的G/G期和部分G/M期细胞周期阻滞。用rhEndostatin处理的HSFs中p53、p27、CDK、细胞周期蛋白D和PCNA的表达水平显著降低。总之，rhEndostatin通过细胞周期的G/G期和/或G/M期阻滞抑制HSFs增殖，这部分归因于细胞周期蛋白D、CDK和PCNA的下调。这些发现表明，rhEndostatin可能通过抑制HSFs增殖在体内减少瘢痕肥厚，可能是一种治疗HS的新型药物。
