| 摘要: |
| 【目的】探究miR-370-3p在皮肤创面愈合中的作用，明确其能否作为miRNA治疗皮肤异常愈合的潜在靶点。【方法】将24只雄性C57BL/6小鼠随机均分为miR-370-3p干预组（miRNA组）、阴性对照组（NC组）和空白对照组（CON组），麻醉后建立8 mm背部皮肤全层缺损模型，miRNA组和NC组小鼠创面消毒后分别单次涂抹miR-370-3p模拟物（miR-370-3p-agomiR）和阴性对照模拟物0.02 mg/只，CON组小鼠创面只进行消毒处理 。 建模后0，7及14 d，观察记录创面愈合情况，统计愈合率。建模后7和14 d，取创面及周围组织样本，部分组织样本用40 g/L多聚甲醛溶液固定，制作切片，进行病理组织学分析 。 采用TargetScan、miRWalk及miRDB软件预测miR-370-3p的靶基因，对靶基因进行KEGG通路富集分析 。 取组织样本，利 用Western blot和RT-qPCR技术检测Ⅰ型胶原蛋白（COLⅠ）和α-平滑肌肌动蛋白（α-SMA）表达水平，以及TGF-β信号通路关键分子（TGF-β1、Smad3、Smad4、Smad7）的基因与蛋白表达水平。【结果】与CON组、NC组相比，miRNA组创面愈合速度变慢，愈合率显著降低（P<0.05），炎症反应较强；建模后7 d新生胶原纤维占比极显著下降，炎症细胞增多，COL Ⅰ和α-SMA表达水平显著降低（P<0. 05）；建模后14 d新生胶原纤维增多，但排列紊乱；CD31阳性血管密度显著减少（P<0. 05）。miR-370-3p靶基因KEGG富集分析发现，TGF-β信号通路与皮肤伤口愈合相关。RT-qPCR结果显示，miRNA组TGF-β信号通路中的TGF-β1、Smad3、Smad4表达水平显著或极显著下降，而负反馈抑制因子Smad7表达水平显著升高（P<0. 05）；Western blot结果证实，miRNA组TGF-β1、Smad3、Smad4、Smad7蛋白的表达水平均显著或极显著降低（P<0.05）。【结论】miR-370-3p可通过抑制TGF-β信号通路，影响胶原沉积、组织收缩及新生血管生成，进而延缓皮肤创面愈合。研究结果提示，miR-370-3p可作为调控动物皮肤创伤修复的潜在靶点。 |
| 关键词: miR-370-3p TGF-β信号通路 创面愈合 胶原重塑 小鼠 |
| DOI：10. 13207/j. jnwafu. 2026. 10. 002 |
| 分类号: |
| 基金项目:吉林省科技发展计划项目（YDZJ202501ZYTS564）；国家自然科学基金项目（C180101） |
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| Effects of miR⁃370⁃3p inhibiting the TGF⁃β pathway on delaying skin wound healing |
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WANG Yan1, WANG Wei1, CHEN Shifan1, ZHANG Bo1, FAN Wei1, TANG Xiukai1, ZHANG Xinyu1, SUN Fuliang1,2
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1.College of Agriculture，Yanbian University，Yanji，Jilin 133002，China;2.Laboratory Animal Center，Yanbian University，Yanji，Jilin 133002，China
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| Abstract: |
| 【Objective】This research aims to investigate the role of miR-370-3p in skin wound healing，to determine its potential as a therapeutic target for abnormal healing in skin treatment.【Method】Twenty-four male C57BL/6 mice were randomly and equally divided into three groups：the miR-370-3p intervention group （miRNA group），the negative control group（NC group），and the blank control group（CON group）.After anesthesia，an 8 mm full-thickness dorsal skin defect model was created. After wound disinfection，mice in the miRNA group and NC group received a single topical application of miR-370-3p-agomiR and negative control mimic at a dose of 0. 02 mg per mouse. Mice in the CON group received wound disinfection only. The wound healing condition was observed and the healing rate was calculated on day 0，7，and 14 post-modeling. On day 7 and 14，wound tissue samples were collected. Some of the samples were fixed in 40 g/L paraformaldehyde for sectioning and histopathological analysis. TargetScan，miRWalk，and miRDB were used to predict the target genes of miR-370-3p，followed by KEGG pathway enrichment analysis. The expression levels of Collagen I
（COL I），and α-smooth muscle actin（α-SMA），and the gene and protein expression levels of key molecules of the TGF-β signaling pathway（TGF-β1，Smad3，Smad4，and Smad7）in tissue samples were detected by using
Western blot and RT-qPCR.【Result】Compared to the CON and NC groups，wound healing of the mice in the miRNA group was markedly delayed，with a significantly reduced healing rate（P<0.05），and a stronger inflam-matory response. On day 7，the proportion of newly formed collagen fibers was significantly decreased，inflamma-tory cell infiltration was increased，and the expression levels of COL I and α -SMA were significantly reduced （P<0.05）.On day 14，the amount of newly formed collagen fibers increased but they were disorganized，and the CD31-positive vessel density was significantly decreased（P<0.05）.Target prediction and KEGG enrich-ment analysis indicated an association between miR-370-3p and the TGF-β signaling pathway in the skin wound healing process. RT-qPCR results showed that the expression levels of TGF- β1，Smad3，and Smad4 in the mice in the miRNA group were significantly or highly significantly downregulated，while the expression level of the negative feedback regulator Smad7 was significantly increased（P<0.05）.However，Western blot analysis indicated that the protein expression levels of TGF- β1，Smad3，Smad4，and Smad7 were all significantly or highly significantly reduced in the mice in miRNA group（P<0.05）.【Conclusion】 miR-370-3p delays skin wound healing by inhibiting the TGF-β signaling pathway，thereby affecting collagen deposition，tissue contraction，and angiogenesis. These findings suggest that miR-370-3p could serve as a potential intervention target for modulating skin wound repair in animals. |
| Key words: miR-370-3p TGF-β signaling pathway wound healing collagen remodeling mice |
