SMAD3在声带损伤中对纤维增生的表达及调控
SMAD3 expression and regulation of fibroplasia in vocal fold injury.
作者信息
Hiwatashi Nao, Benedict Peter A, Dion Gregory R, Bing Renjie, Kraja Iv, Amin Milan R, Branski Ryan C
机构信息
NYU Voice Center, Department of Otolaryngology-Head and Neck Surgery, New York University School of Medicine, New York, New York, U.S.A.
出版信息
Laryngoscope. 2017 Sep;127(9):E308-E316. doi: 10.1002/lary.26648. Epub 2017 May 20.
OBJECTIVE
Recent reports highlight the efficacy of small interfering RNA (siRNA) targeting SMAD3 to regulate transforming growth factor β (TGF-β)-mediated fibroplasia in vocal fold fibroblasts. The current study sought to investigate SMAD3 expression during wound healing in vivo and quantify the downstream transcriptional events associated with SMAD3 knockdown in vitro.
STUDY DESIGN
In vivo and in vitro.
METHODS
Unilateral vocal fold injury was created in a rabbit model. SMAD3 and SMAD7 mRNA expression was quantified at 1 hour and 1, 3, 7, 14, 30, 60, and 90 days following injury. In vitro, multi-gene analysis technology was employed in our immortalized human vocal-fold fibroblast cell line following TGF-β1 stimulation ± SMAD3 knockdown across time points.
RESULTS
SMAD3 mRNA expression increased following injury; upregulation was significant at 3 and 7 days compared to control (both P < 0.001). SMAD7 mRNA was also upregulated at 3, 7, and 14 days (P = 0.02, P < 0.001, and P < 0.001, respectively). In vitro, SMAD3 knockdown reduced the expression of multiple profibrotic, TGF-β signaling, and extracellular matrix metabolism genes at 6 and 24 hours following TGF-β1 stimulation.
CONCLUSION
Cumulatively, these data support SMAD3 as a potential master regulator of TGF-β-mediated fibrosis. SMAD3 transcription peaked 7 days following injury. Multi-gene analysis indicated that the therapeutic effectiveness of SMAD3 knockdown may be related to regulation of downstream mediators of fibroplasia and altered TGF-β signaling.
LEVEL OF EVIDENCE
NA. Laryngoscope, 127:E308-E316, 2017.
目的
近期报道强调了靶向SMAD3的小干扰RNA（siRNA）在调节声带成纤维细胞中转化生长因子β（TGF-β）介导的纤维增生方面的功效。本研究旨在调查体内伤口愈合过程中SMAD3的表达情况，并在体外定量与SMAD3基因敲低相关的下游转录事件。
研究设计
体内和体外研究。
方法
在兔模型中制造单侧声带损伤。在损伤后1小时以及1、3、7、14、30、60和90天对SMAD3和SMAD7 mRNA表达进行定量。在体外，在永生化的人声带成纤维细胞系中，在TGF-β1刺激后±SMAD3基因敲低的不同时间点采用多基因分析技术。
结果
损伤后SMAD3 mRNA表达增加；与对照组相比，在第3天和第7天上调显著（均P<0.001）。SMAD7 mRNA在第3、7和14天也上调（分别为P = 0.02、P<0.001和P<0.001）。在体外，TGF-β1刺激后6小时和24小时，SMAD3基因敲低降低了多种促纤维化、TGF-β信号传导和细胞外基质代谢基因的表达。
结论
总体而言，这些数据支持SMAD3作为TGF-β介导的纤维化的潜在主要调节因子。SMAD3转录在损伤后7天达到峰值。多基因分析表明，SMAD3基因敲低的治疗效果可能与调节纤维增生的下游介质以及改变TGF-β信号传导有关。
证据水平
无。《喉镜》，2017年，第127卷，E308 - E316页
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