Wang X, Zhang Y, Jiang B H, Zhang Q, Zhou R P, Zhang L, Wang Chen
Department of Dermatology and Dermatologic Surgery, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, 639 Zhi Zao Ju Road, Shanghai 200011, PR China.
Department of Orthopedics, Shanghai Tongji Hospital, Tongji University School of Medicine, 389 Xincun Road, Shanghai 200065, PR China.
Exp Cell Res. 2017 Dec 15;361(2):201-209. doi: 10.1016/j.yexcr.2017.09.009. Epub 2017 Oct 19.
Hypertrophic scar (HS) formation is associated with the fibrosis of fibrocytes caused by excessive extracellular matrix (ECM) synthesis and deposition, the initial event of HS formation. Our high throughput screen of miRNA expression profiles identified hsa-miR31-5p, whose transcription level was most differentially in normal skin fibroblasts (NS) and HS among other miRNAs. The level of hsa-miR31-5p in HS was significantly higher than in NS. In-vitro functional experiments showed hsa-miR31-5p knockdown remarkably suppressed the proliferation of hypertrophic scar fibroblasts (HSFBs) under hypoxia, promoted cell invasion, and inhibited the expression of Collagen I and III and Fibronectin (FN), suggesting that hsa-miR31-5p knockdown effectively reduces HS formation caused by excessive ECM synthesis and deposition in HSFBs under hypoxia. Mechanism study showed that the regulation of HS formation by hsa-miR31-5p was mediated by its target gene, factor-inhibiting HIF-1 (FIH): under hypoxia, hsa-miR31-5p down-regulated FIH and thus increased the level of hypoxia inducible factor-1α (HIF-1α), which subsequently activated the HIF-1α fibrosis regulation pathway in HSFBs, and stimulated the proliferation and ECM synthesis in HSFBs, eventually resulting in fibrosis and scar formation. The data also show that knockdown of hsa-miR31-5p in HSFBs impaired the trend of increased proliferation, reduced invasion and excessive ECM synthesis and deposition caused by HIF-1a activation under hypoxia through upregulating FIH, indicating that knockdown of hsa-miR31-5p effectively inhibits the formation of HS. In conclusion, hsa-miR31 -5p plays an important role in HS formation by inhibiting FIH and regulating the HIF-1α pathway. Therefore, hsa-miR31 -5p may be a novel therapeutic target for HS.
增生性瘢痕(HS)的形成与细胞外基质(ECM)过度合成和沉积所导致的纤维细胞纤维化有关,这是HS形成的初始事件。我们对微小RNA(miRNA)表达谱进行的高通量筛选鉴定出了hsa-miR31-5p,在正常皮肤成纤维细胞(NS)和HS中,其转录水平与其他miRNA相比差异最为显著。HS中hsa-miR31-5p的水平显著高于NS。体外功能实验表明,敲低hsa-miR31-5p可显著抑制缺氧条件下增生性瘢痕成纤维细胞(HSFBs)的增殖,促进细胞侵袭,并抑制I型和III型胶原蛋白以及纤连蛋白(FN)的表达,这表明敲低hsa-miR31-5p可有效减少缺氧条件下HSFBs中因ECM过度合成和沉积而导致的HS形成。机制研究表明,hsa-miR31-5p对HS形成的调节是由其靶基因——缺氧诱导因子-1抑制因子(FIH)介导的:在缺氧条件下,hsa-miR31-5p下调FIH,从而增加缺氧诱导因子-1α(HIF-1α)的水平,随后激活HSFBs中的HIF-1α纤维化调节途径,并刺激HSFBs的增殖和ECM合成,最终导致纤维化和瘢痕形成。数据还显示,在HSFBs中敲低hsa-miR31-5p可通过上调FIH来削弱缺氧条件下HIF-1α激活所导致的增殖增加、侵袭减少以及ECM过度合成和沉积的趋势,这表明敲低hsa-miR31-5p可有效抑制HS的形成。总之,hsa-miR31-5p通过抑制FIH并调节HIF-1α途径在HS形成中发挥重要作用。因此,hsa-miR31-5p可能是HS的一个新的治疗靶点。