MiR-486-5p/FOXO1轴对深二度烧伤创面修复过程中角质形成细胞增殖和迁移的调节作用

doi:10.3969/j.issn.1007-3205.2023.07.006

河北医科大学学报 ›› 2023, Vol. 44 ›› Issue (7): 773-780,797.doi: 10.3969/j.issn.1007-3205.2023.07.006

MiR-486-5p/FOXO1轴对深二度烧伤创面修复过程中角质形成细胞增殖和迁移的调节作用

陕西省榆林市第二医院烧伤整形手足外科，陕西榆林 719000

出版日期:2023-07-25 发布日期:2023-07-24
作者简介:高富成（1989-），男，陕西延安人，陕西省榆林市第二医院主治医师，医学硕士，从事烧伤疾病诊治研究。
基金资助:
陕西省自然科学基金（20190529）

MiR-486-5p/FOXO1 axis regulates keratinocyte proliferation and migration during repair of deep second-degree burn wound

Department of Hand and Foot Surgery of Burn and Plastic Surgery, the Second Hospital of Yulin City, Shaanxi Province, Yulin 719000, China

Online:2023-07-25 Published:2023-07-24

摘要/Abstract

摘要： 目的探讨miR-486-5p/叉头框转录因子O亚族1（forkhead box transcription factor O subfamily 1,FOXO1）轴对深二度烧伤创面修复过程中角质形成细胞的增殖和迁移的调节机制。
方法小鼠建立深二度烧伤模型，随机分为NC mimic+Vector组、FOXO1组、miR-486-5p mimic组和miR-486-5p mimic+FOXO1组。采用miR-486-5p mimic和（或）FOXO1处理伤面，观察伤面愈合情况。在烧伤后分别于不同时间点［止血（0 d）、炎症（1 d）和增殖（7 d和14 d）］采用原位杂交技术和实时荧光定量逆转录聚合酶链反应（reverse transcription quantitative polymerase chain reaction，RT-qPCR）
法检测伤面边缘组织中miR-486-5p表达部位和水平，免疫荧光染色法检测FOXO1表达。通过荧光素酶报告分析FOXO1与miR-486-5p结合关系。构建miR-486-5p过表达或敲低人永生化角质形成细胞（human HaCaTKeratinocytes，HaCaT）模型，通过EdU法和划痕试验考察细胞的增殖和迁移情况，Western blot法分析细胞中FOXO1蛋白表达。
结果 miR-486-5p在小鼠深二度烧伤伤面愈合过程中显著上调。体内模型显示，miR-486-5p通过抑制FOXO1表达促进伤面愈合。在HaCaT细胞中，miR-486-5p过表达增加了细胞的增殖和迁移，抑制miR-486-5p则具有相反的效果。FOXO1是角质形成细胞中miR-486-5p的直接靶标。FOXO1过表达导致HaCaT细胞的EdU阳性细胞的百分比和伤面愈合降低，并且逆转了miR-486-5p对EdU阳性细胞和伤面愈合的诱导作用。
结论 miR-486-5p是深二度烧伤创面修复的关键调节剂，通过抑制FOXO1表达促进角质形成细胞的增殖和迁移。

关键词: 烧伤, miR-486-5p, 叉头框转录因子O亚族1

Abstract: Objective To explore the regulatory mechanism of the miR-486-5p/forkhead box transcription factor O subfamily 1 (FOXO1) axis on the proliferation and migration of keratinocytes during repair of deep second-degree burn wound.
Methods A mouse model of deep second-degree burn was established, and the mice were randomly divided into 4 groups: NC mimic+Vector group, FOXO1 group, miR-486-5p mimic group and miR-486-5p mimic+FOXO1 group. The wound was treated with miR-486-5p mimic and/or FOXO1 to observe the wound healing. At different time points after burn, including hemostasis (0 h), inflammation (24 h), and proliferation (7th and 14th day), in situ hybridization and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were used to detect miR-486-5p expression sites and levels in wound edge tissues, respectively. The expression of FOXO1 was detected by immunofluorescence staining. Luciferase reporter analysis confirmed the binding relationship between FOXO1 and miR-486-5p. The human HaCaTkeratinocytes(HaCaT) cell model of miR-486-5p overexpression or knockdown was constructed. The proliferation and migration of cells were investigated by EdU method and scratch test, and the expression of FOXO1 protein in cells was analyzed by Western blot.
Results miR-486-5p was significantly up-regulated during the healing of deep second-degree burn wounds in mice. In vivo model showed that miR-486-5p promoted wound healing by inhibiting the expression of FOXO1. In HaCaT cells, miR-486-5p overexpression increased proliferation and migration of the cells, where as miR-486-5p inhibition had the reverse effect. FOXO1 was a direct target of miR-486-5p in keratinocytes. FOXO1 overexpression decreased the percentage of EdU positive cells and wound healing of HaCaT cells, and reversed the induction of EdU positive cells and wound healing by miR-486-5p.
Conclusion MiR-486-5p is a crucial regulator in repair of deep second-degree burn wound that facilitates keratinocyte proliferation and migration by inhibiting the expression of FOXO1.

Key words: burns, miR-486-5p, forkhead box transcription factor O subfamily 1

高富成, 王耀军, 薛佳杰, 郝青晔, 高登文, 郭雷. MiR-486-5p/FOXO1轴对深二度烧伤创面修复过程中角质形成细胞增殖和迁移的调节作用 [J]. 河北医科大学学报, 2023, 44(7): 773-780,797.

GAO Fu-cheng, WANG Yao-jun, XUE Jia-jie, HAO Qing-ye, GAO Deng-wen, GUO Lei. MiR-486-5p/FOXO1 axis regulates keratinocyte proliferation and migration during repair of deep second-degree burn wound [J]. Journal of Hebei Medical University, 2023, 44(7): 773-780,797.

[1]	徐琦量, 陈晓明, 戴强, 施文娟, 周剑, 蒋逸. 血乳酸、降钙素原清除率对烧伤休克患者预后的预测价值 [J]. 河北医科大学学报, 2023, 44(8): 941-944,951.
[2]	王旭文, 李亮, 苏伟海, 齐杰, 生孟军. 重组人酸性成纤维细胞生长因子联合夫西地酸乳膏局部应用对深Ⅱ度烧伤患者创面愈合、炎症水平、疼痛介质的影响 [J]. 河北医科大学学报, 2023, 44(6): 681-685,691.
[3]	何静, 徐刚, 姜海, 刘宇, 刘冬梅. 双层人工真皮移植在关节深度烧伤创面的早期修复效果[J]. 河北医科大学学报, 2021, 42(10): 1166-1170.
[4]	张莉, 王莉, 田长征, 郝雪莲. 右美托咪定对重度烧伤患者血浆IL-1β、IL-6、血清S-100β水平及术后谵妄的影响[J]. 河北医科大学学报, 2020, 41(11): 1344-1348.
[5]	王丽娇，谢颖，胡环宇，杜红珍，骆彬*，李增宁. 应用益生菌制剂对严重烧伤患者炎症反应的影响[J]. 河北医科大学学报, 2019, 40(12): 1426-1429.
[6]	石〓聪，王东凯，耿志辉*. 龙血竭联合重组牛碱性成纤维细胞生长因子凝胶治疗烧烫伤临床研究[J]. 河北医科大学学报, 2018, 38(10): 1217-1220.
[7]	聂〓婵1，王〓涛1，施尚鹏1，邓呈亮2，石修权1，3*. 烧伤对儿童伤残率和瘢痕率的影响分析#br#[J]. 河北医科大学学报, 2018, 38(1): 58-63.
[8]	徐丽红. FGF+Collagen联合百多邦治疗烧伤后残余创面对炎性因子表达及创面愈合的影响[J]. 河北医科大学学报, 2017, 37(10): 1168-1171.
[9]	邵洪波;姚月明;魏伟. 碱烧伤后血液流变学的实验研究[J]. , 2015, 36(7): 821-821.
[10]	李娟;任丽青;张翠娜;白永强. 护理干预对烧伤患者出院后抗瘢痕行为能力的影响[J]. , 2014, 35(7): 765-765.
[11]	王车江;张庆富;吴泳泽;冯建科;白永强;邵洪波. 高压电烧伤大鼠坐骨神经微循环灌流量变化及己酮可可碱的干预作用[J]. , 2013, 34(9): 1023-1023.
[12]	于蕾;张久新. 重度烧伤休克期68例输液管理分析[J]. , 2013, 34(9): 0-0.
[13]	李平;周长浩;魏砚砚. 枸橼酸舒芬太尼超前镇痛对大面积烧伤患者切削痂植皮术术后镇痛的影响[J]. , 2013, 34(8): 972-972.
[14]	陈磊. 新鲜羊膜移植联合肝素在角膜烧伤治疗中的应用[J]. , 2012, 33(5): 531-531.
[15]	王车江;张桂华;刘洪霞;张庆富. 特重度烧伤合并Coombs阳性1例[J]. , 2012, 33(5): 0-0.

MiR-486-5p/FOXO1轴对深二度烧伤创面修复过程中角质形成细胞增殖和迁移的调节作用

MiR-486-5p/FOXO1 axis regulates keratinocyte proliferation and migration during repair of deep second-degree burn wound

HTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价