中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (8): 1609-1617.doi: 10.12307/2025.341

• 皮肤粘膜组织构建 skin and mucosal tissue construction • 上一篇    下一篇

miR-27a-3p激活MAPK信号通路促进人增生性瘢痕成纤维细胞的增殖

李  俊1，巩晶晶1，孙国斌1，郭  睿1，丁  杨1，强立娟1，张晓莉1，方占海2   

  1. 宁夏回族自治区人民医院(宁夏医科大学附属自治区人民医院)，1烧伤整形外科，2神经外科，宁夏回族自治区银川市  750004
  • 收稿日期:2024-01-10 接受日期:2024-04-03 出版日期:2025-03-18 发布日期:2024-07-05
  • 通讯作者: 方占海，副教授，宁夏回族自治区人民医院(宁夏医科大学附属自治区人民医院)神经外科，宁夏回族自治区银川市 750004
  • 作者简介:李俊，女，1973年生，宁夏回族自治区银川市人，汉族，主任医师，主要从事增生性瘢痕形成的机制研究。
  • 基金资助:
    宁夏自然科学基金项目(2023AAC03456)，项目负责人：郭睿

miR-27a-3p promotes the proliferation of human hypertrophic scar fibroblasts by regulating mitogen-activated protein kinase signaling pathway

Li Jun1, Gong Jingjing1, Sun Guobin1, Guo Rui1, Ding Yang1, Qiang Lijuan1, Zhang Xiaoli1, Fang Zhanhai2    

  1. 1Department of Burn and Plastic Surgery, 2Department of Neurosurgery, People’s Hospital of Ningxia Hui Autonomous Region (Ningxia Medical University Affiliated Autonomous Region People’s Hospital), Yinchuan 750004, Ningxia Hui Autonomous Region, China 
  • Received:2024-01-10 Accepted:2024-04-03 Online:2025-03-18 Published:2024-07-05
  • Contact: Fang Zhanhai, Associate professor, Department of Neurosurgery, People’s Hospital of Ningxia Hui Autonomous Region (Ningxia Medical University Affiliated Autonomous Region People’s Hospital), Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Li Jun, Chief physician, Department of Burn and Plastic Surgery, People’s Hospital of Ningxia Hui Autonomous Region (Ningxia Medical University Affiliated Autonomous Region People’s Hospital), Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    Natural Science Foundation of Ningxia Hui Autonomous Region, No. 2023AAC03456 (to GR)

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摘要：


文题释义：
微小RNA(miRMA)：广泛存在于真核细胞的细胞质中，长度为18-22个核苷酸，miRNA在机体的细胞分化、生长发育、凋亡等一系列生命活动中发挥重要作用。
增殖：是细胞的基本生命特征，主要以分裂的方式进行。

背景：目前多项研究证实了丝裂原活化蛋白激酶信号通路参与了细胞的增殖过程，且miRNA参与增生性瘢痕的发生发展，因此深入探讨了miR-27a-3p和丝裂原活化蛋白激酶信号通路在病理性瘢痕形成中的作用。
目的：探究miR-27a-3p通过丝裂原活化蛋白激酶信号通路对人增生性瘢痕成纤维细胞增殖的影响。
方法：收集皮肤标本并分别分离出原代成纤维细胞，倒置显微镜观察原代细胞，免疫荧光予以验证；采用qRT-PCR检测miR-27a-3p在组织中的相对表达水平；利用数据库预测miR-27a-3p的靶基因，再将预测的靶基因进行基因本体功能富集分析及京都基因与基因组百科全书生物通路富集分析；设置分组为：空白对照组、阴性对照组、miR-27a-3p过表达组、miR-27a-3p抑制组、miR-27a-3p过表达+p38 丝裂原活化蛋白激酶抑制剂组、miR-27a-3p过表达+细胞外信号调节蛋白激酶抑制剂组、miR-27a-3p过表达+c-Jun氨基末端激酶抑制剂组，Western blot检测细胞外调节蛋白激酶、c-Jun氨基末端激酶和p38激酶总量及其磷酸化水平，采用CCK-8法和EdU检测细胞增殖情况。
结果与结论：①与正常皮肤成纤维细胞相比，增生性瘢痕成纤维细胞的增殖活性更强(P < 0.05)，增殖速度也更快(P < 0.001)；②与正常皮肤相比，miR-27a-3p在增生性瘢痕中呈高表达(P < 0.001)；③与阴性对照组相比，过表达miR-27a-3p能促进细胞的增殖活性(P < 0.001)和增殖水平(P < 0.001)；④与阴性对照组相比，敲低miR-27a-3p能抑制细胞的增殖活性(P < 0.05)和增殖水平(P < 0.001)；⑤与阴性对照组相比，过表达miR-27a-3p促进细胞外调节蛋白激酶、c-Jun氨基末端激酶和p38丝裂原活化蛋白激酶的磷酸化水平(P < 0.05)；与阴性对照组相比，敲减miR-27a-3p能抑制细胞外调节蛋白激酶、c-Jun氨基末端激酶和p38丝裂原活化蛋白激酶的磷酸化水平(P < 0.05)；⑥与miR-27a-3p过表达组相比，细胞外调节蛋白激酶、c-Jun氨基末端激酶和p38丝裂原活化蛋白激酶的特异性抑制剂可逆转miR-27a-3p对成纤维细胞的增殖活性(P < 0.01)和增殖水平(P < 0.001)；⑦提示miR-27a-3p通过激活丝裂原活化蛋白激酶信号通路促进人增生性瘢痕成纤维细胞的增殖。
https://orcid.org/0009-0005-7263-2212(李俊)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: miR-27a-3p, 丝裂原活化蛋白激酶, 增生性瘢痕, 成纤维细胞, 增殖

Abstract: BACKGROUND: Multiple studies have confirmed that mitogen-activated protein kinase (MAPK) signaling pathway is involved in cell proliferation, and microRNA (miR) is involved in the occurrence and development of hypertrophic scars. Therefore, the role of miR-27a-3p and MAPK signaling pathways in pathological scar formation has been further explored.
OBJECTIVE: To explore the effect of miR-27a-3p on the proliferation of human hypertrophic scar fibroblasts through the MAPK signaling pathway.
METHODS: The primary fibroblasts were isolated and collected from the skin samples. The primary fibroblasts were observed by inverted microscope and verified by immunofluorescence. The relative expression level of miR-27a-3p in tissues was detected by qRT-PCR. The target genes of hsa-miR-27a-3p were predicted using the database, and then the predicted target genes were enriched by gene ontology function analysis and biological pathway enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes. There were seven groups: blank control, negative control, miR-27a-3p mimic, miR-27a-3p inhibitor, miR-27a-3p mimic+p38 MAPK inhibitor, miR-27a-3p mimic+extracellular regulated protein kinase inhibitor, miR-27a-3p mimic+c-Jun N-terminal kinase inhibitor. Western blot was used to detect the levels of extracellular regulated protein kinase, c-Jun N-terminal kinase inhibitor. and p38 kinase and their phosphorylation levels. Cell counting kit-8 and EdU were used to detect cell proliferation.
RESULTS AND CONCLUSION: Compared with normal skin fibroblasts, hypertrophic scar fibroblasts had stronger proliferative activity (P < 0.05) and faster proliferation level (P < 0.001). Compared with normal skin, miR-27a-3p was highly expressed in hypertrophic scars (P < 0.001). Compared with the negative control group, overexpression of miR-27a-3p could promote cell proliferation activity (P < 0.001) and proliferation levels (P < 0.001). Compared with the negative control group, knockdown of miR-27a-3p could inhibit the proliferation activity (P < 0.05) and proliferation levels (P < 0.001). Compared with the negative control group, overexpression of miR-27a-3p promoted the phosphorylated levels of extracellular regulated protein kinase, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinase (P < 0.05). Compared with the negative control group, knockdown of miR-27a-3p inhibited the phosphorylated levels of extracellular regulated protein kinase, c-Jun N-terminal kinase, and p38 MAPK (P < 0.05). Compared with the miR-27a-3p mimic group, specific inhibitors of extracellular regulated protein kinase, c-Jun N-terminal kinase, and p38 MAPK reversed the effects of miR-27a-3p on the proliferative activity (P < 0.01) and proliferation level (P < 0.001) of fibroblasts. To conclude, these results suggest that miR-27a-3p promotes the proliferation of human hypertrophic scar fibroblasts by activating the MAPK signaling pathway.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: miR-27a-3p, mitogen-activated protein kinase, hypertrophic scar, fibroblast, proliferation

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