Fang Chih-Yuan, Chen Yin-Ju, Lin Kuan-Chou, Chen Yueh-Hsin, Yang Kai-Chiang
School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan.
Division of Oral and Maxillofacial Surgery, Department of Dentistry, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan.
J Oral Pathol Med. 2025 Jun 27. doi: 10.1111/jop.70009.
Oral submucous fibrosis is characterized by excessive collagen deposition and is highly associated with a patient's betel nut chewing habit. Arecoline initiates the transforming growth factor-beta (TGF-β)/Smads signaling pathway and activates downstream fibrotic genes. Dysregulation of microRNA (miR) expression is involved in the OSF progression, and miR modulation is a promising treatment. As one miR can target multiple mRNAs, and one mRNA has multiple binding sites to different miRs, we thus propose that simultaneous co-transfection of anti-fibrotic miRs may have a better therapeutic effect than single miR transfection.
Human oral fibroblasts were first subjected to arecoline stimulation and then transfected with 16 miRs individually. Based on the ability to downregulate TGFB1 and actin alpha 2, smooth muscle (ACTA2) mRNA levels, the miR-29a-3p mimic, miR-196a-3p mimic, and miR-509-5p mimic were selected for co-transfection.
In addition to downregulation of collagen type I alpha 1 chain (COL1A1), COL3A1, COL5A1, matrix metalloproteinase-1 (MMP1), MMP7, tissue inhibitor of metalloproteinase-1 (TIMP1), and TIMP2 mRNA expressions, co-transfection with the three miRs led to a more significant downregulation of COL1A1 and MMP1 expressions. A Western blot analysis revealed that co-transfection of the miRs efficiently suppressed the TGF-β/Smads pathway and extracellular matrix component productions. Furthermore, co-transfection with miRs more effectively inhibited wound closure and collagen gel contraction compared to single miR transfection.
Co-transfection of anti-fibrotic miRs can be a promising treatment for oral submucous fibrosis.
口腔黏膜下纤维化的特征是胶原蛋白过度沉积，且与患者咀嚼槟榔的习惯高度相关。槟榔碱启动转化生长因子-β（TGF-β）/Smads信号通路并激活下游纤维化基因。微小RNA（miR）表达失调参与口腔黏膜下纤维化的进展，miR调节是一种有前景的治疗方法。由于一个miR可以靶向多个mRNA，且一个mRNA有多个与不同miR的结合位点，因此我们提出，与单个miR转染相比，同时共转染抗纤维化miR可能具有更好的治疗效果。
首先将人口腔成纤维细胞用槟榔碱刺激，然后分别用16种miR转染。基于下调TGFB1和α2平滑肌肌动蛋白（ACTA2）mRNA水平的能力，选择miR-29a-3p模拟物、miR-196a-3p模拟物和miR-509-5p模拟物进行共转染。
除了下调I型胶原蛋白α1链（COL1A1）、COL3A1、COL5A1、基质金属蛋白酶-1（MMP1）、MMP7、金属蛋白酶组织抑制剂-1（TIMP1）和TIMP2的mRNA表达外，这三种miR的共转染导致COL1A1和MMP1表达的下调更为显著。蛋白质印迹分析显示，miR的共转染有效抑制了TGF-β/Smads通路和细胞外基质成分的产生。此外，与单个miR转染相比，miR的共转染更有效地抑制了伤口闭合和胶原凝胶收缩。
抗纤维化miR的共转染可能是口腔黏膜下纤维化的一种有前景的治疗方法。
