| 摘要: |
| 目的 探究瘢痕表皮细胞在低能量CO2点阵激光改善烧伤后瘢痕中的作用及可能的分子机制。方法 建立大鼠背部大面积烧伤后瘢痕模型。对3只烧伤后瘢痕模型大鼠给予30 mJ低能量CO2点阵激光干预，观察瘢痕表皮细胞活化情况，分离表皮组织行转录组测序筛选激活通路。将18只烧伤后瘢痕模型大鼠随机分为3组（n＝6）：对照组不予激光干预，激光组予30 mJ CO2点阵激光干预，激光＋抑制剂组予30 mJ CO2点阵激光干预＋IWR-1（Wnt/β-联蛋白通路抑制剂）瘢痕内注射，以验证筛选到的通路激活情况及其效应。通过H-E染色、马松染色和蛋白质印迹法检测表皮细胞和成纤维细胞活化、Wnt/β-联蛋白通路激活及瘢痕改善情况。结果 低能量激光干预后，烧伤后瘢痕模型大鼠瘢痕表皮组织Ki67、增殖细胞核抗原（PCNA）、细胞角蛋白19（CK19）、p63阳性细胞数量增多，活化明显；转录组测序结果结合文献分析筛选出Wnt/β-联蛋白通路作为候选通路。在验证性实验中，与对照组相比，激光干预后第5天烧伤后瘢痕模型大鼠瘢痕表皮细胞Wnt/β-联蛋白通路被激活，激光干预后第30天大鼠瘢痕组织真皮胶原排列更为疏松、真皮厚度变薄、α-平滑肌肌动蛋白阳性成纤维细胞数减少、Ⅰ型和Ⅲ型胶原蛋白的含量及Ⅰ型/Ⅲ型胶原蛋白的比例下降；给予Wnt/β-联蛋白通路抑制剂后，上述低能量激光干预诱导的Wnt/β-联蛋白通路激活、瘢痕表皮细胞活化和瘢痕改善现象均被逆转。结论 低能量CO2点阵激光可通过激活瘢痕表皮细胞Wnt/β-联蛋白通路活化瘢痕表皮细胞、改善烧伤后瘢痕。 |
| 关键词: 烧伤 瘢痕 低能量点阵激光 表皮细胞 Wnt/β-联蛋白通路 |
| DOI：10.16781/j.CN31-2187/R.20240280 |
| 投稿时间：2024-04-30修订日期：2024-06-02 |
| 基金项目:国家自然科学基金（81930057，82272260）;军队后勤科研重点项目（JKBWS23C1018，CHJ23C031）;上海市重中之重研究中心建设项目（2023ZZ02013）. |
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| Low-energy CO2 fractional laser treatment for post-burn scars via activating Wnt/β-catenin pathway in scar epithelial cells in rats |
| GU Haoyu,LIU Yingying,YANG Lu,XIAO Shichu,LUO Pengfei*,XIA Zhaofan* |
(Department of Burn Surgery, The First Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 200433, China
*Corresponding authors) |
| Abstract: |
| Objective To investigate the role of scar epithelial cells and its potential molecular mechanisms in the efficacy of low-energy CO2 fractional laser treating post-burn scars. Methods The model of post-major burn scars on the back of rat was established. Three rats with post-major burn scars received 30 mJ low-energy CO2 fractional laser treatment to detect the activation of scar epidermal cells. Epidermal tissue of scars was isolated for RNA sequencing to screen activated pathways. Subsequently, 18 rats with post-major burn scars were randomly divided into 3 groups (n＝6): the control group without laser treatment, the laser group receiving 30 mJ CO2 fractional laser treatment, and the laser＋inhibitor group receiving laser treatment and intra-scar injection of IWR-1 (a Wnt/β-catenin pathway inhibitor), to verify the activation status and effects of the selected pathways. Hematoxylin-eosin staining, Masson staining, and Western blotting were used to detect the proliferation of epithelial cells and fibroblasts, the activation of Wnt/β-catenin pathway, as well as the improvement of scar profiles. Results After low-energy laser treatment, there was a significant increase in the number of Ki67-positive, proliferating cell nuclear antigen (PCNA)-positive, cytokeratin 19 (CK19)-positive, and p63-positive cells in the scar epithelial tissue. RNA sequencing coupled with literature analysis identified Wnt/β-catenin pathway as a potential candidate pathway. In the confirmatory experiment, compared to the control group, the Wnt/β-catenin pathway was activated in scar epithelial cells in the laser group 5 d post-laser intervention. After 30 d laser intervention, dermal collagen exhibited a more loosened arrangement, with reduced dermal thickness and significantly less α-smooth muscle actin (α-SMA)-positive fibroblasts compared to the control group. CollagenⅠ, collagen Ⅲ, and the relative ratio of collagen Ⅰ to Ⅲ in the laser group were at a lower level than those in the control group. Administration of the Wnt/β-catenin pathway inhibitor blocked the activation of the Wnt/β-catenin pathway induced by low-energy laser, the proliferation of scar epithelial cells and the improvement of scar profiles. Conclusion Low-energy CO2 fractional laser treatment can activate the Wnt/β-catenin pathway of scar epithelial cells, thereby activating epithelial cells and yielding significant scar improvements. |
| Key words: burn scar low-energy fractional laser epidermal cells Wnt/β-catenin pathway |
