NRG2通过调控PI3K-AKT通路促进创面修复相关巨噬细胞的M2极化  

NRG2 Promotes M2 Polarization of Wound Healing-Related Macrophages by Regulating the PI3K-AKT Pathway

作　　者：彭友华[1] 李婧林 张予晋 周蓉 潘意 高贵云[2] PENG You-hua;LI Jing-lin;ZHANG Yu-jin;ZHOU Rong;PAN Yi;GAO Gui-yun(Department of Dermatology,The Second Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Hunan Province,410005,China;Department of Dermatology,Hunan Aerospace Hospital,Hunan Province,410221,China)

机构地区：[1]湖南中医药大学第二附属医院皮肤科,湖南长沙410005 [2]湖南航天医院皮肤科,湖南长沙410221

出　　处：《中国医疗美容》2022年第11期54-57,共4页China Medical Cosmetology

基　　金：湖南省自然科学基金资助(2020JJ5431);湖南中医药管理局重点项目(C2022025)。

摘　　要：目的探讨NRG2对皮肤创面的修复过程及单核巨噬细胞表型转化的影响及具体机制。方法生物信息学分析获得巨噬细胞相关的创面修复关键因子,验证NRG2的差异表达。构建小鼠皮肤创面模型,在模型中验证NRG2 mRNA及蛋白表达水平;构建NRG2过表达质粒,将NRG2过表达质粒及对照质粒转染至THP-1细胞,qPCR及western blot检测转染效率,流式细胞术检测NRG2对巨噬细胞极化的影响;通过NRG2过表达质粒和(或)PI3K-AKT抑制剂LY294002进行干预,Western blot法检测p-PI3K、PI3K、p-AKT、AKT蛋白表达水平,流式细胞术检测巨噬细胞表型。结果数据集GSE157291中663个在创面修复晚期上调的DEGs及444个在创面修复晚期下调的DEGs,其中,NRG2在创面修复晚期显著上调。相较于创面修复早期,创面修复晚期小鼠NRG2 mRNA及蛋白水平显著上调。NRG2过表达质粒可在细胞中成功实现NRG2的过表达。与vector-NC组相比,NRG2组的CD80、86标记的M1巨噬细胞百分率显著降低,p-PI3K、p-AKT表达水平及CD163、CD206标记的M2巨噬细胞百分率显著升高。PI3K-AKT抑制剂LY294002抵消了NRG2的促进作用。结论NRG2通过调控PI3K-AKT通路促进创面修复相关巨噬细胞的M2极化。Objective To explore the healing process of NRG2 on skin wounds and its effect on monocyte-macrophage phenotype transformation and its specific mechanism.Methods Bioinformatics analysis obtained the key factors of macrophage-related wound repair,and verified the differential expression of NRG2.mouse skin wound model was constructed to verify the expression levels of NRG2 mRNA and protein in the model;The NRG2 overexpression plasmid was constructed,and the NRG2 overexpression plasmid and the control plasmid were transfected into THP-1 cells.The transfection efficiency was detected by qPCR and western blot,and the effect of NRG2 on macrophage polarization was detected by flow cytometry.The NRG2 overexpression plasmid and/or PI3K-AKT inhibitor LY294002 were intervened,the protein expression levels of p-PI3K,PI3K,p-AKT and AKT were detected by Western blot,and the expression of macrophages was detected by flow cytometry.Results In the dataset GSE157291,663 DEGs were up-regulated in the late wound healing stage and 444 DEGs were down-regulated in the late wound healing stage.Among them,NRG2 was significantly up-regulated in the late wound healing stage.Compared with the early wound healing stage,the NRG2 mRNA and protein levels were significantly up-regulated in the late wound healing mouse.The NRG2 overexpression plasmid can successfully achieve the overexpression of NRG2 in cells.Compared with the vector-NC group,the percentages of M1 macrophages marked by CD80 and 86 in the NRG2 group were significantly decreased,while the expression levels of p-PI3K and p-AKT and the percentages of M2 macrophages marked by CD163 and CD206 were significantly increased.The PI3K-AKT inhibitor LY294002 counteracted the promotion of NRG2.Conclusion NRG2 promotes M2 polarization of wound healing-related macrophages by regulating the PI3K-AKT pathway.

关 键 词：创面修复 神经调节素2 巨噬细胞 极化 

分 类 号：R641[医药卫生—外科学]