Hypoxia macrophage-derived exosomal miR-26b-5p targeting PTEN promotes the development of keloids.

IF 6.3 1区医学 Q1 DERMATOLOGY Burns & Trauma Pub Date : 2024-02-29 eCollection Date: 2024-01-01 DOI:10.1093/burnst/tkad036

Siya Dai, Mingyuan Xu, Qianqian Pang, Jiaqi Sun, Xiaohu Lin, Xi Chu, Chunyi Guo, Jinghong Xu

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Abstract

Background: Hypoxia is the typical characteristic of keloids. The development of keloids is closely related to the abnormal phenotypic transition of macrophages. However, the role of exosomal microRNAs (miRNAs) derived from hypoxic macrophages in keloids remains unclear. This study aimed to explore the role of hypoxic macrophage-derived exosomes (HMDE) in the occurrence and development of keloids and identify the critical miRNA.

Methods: The expression of CD206⁺ M2 macrophage in keloids and normal skin tissues was examined through immunofluorescence. The polarization of macrophages under a hypoxia environment was detected through flow cytometry. The internalization of macrophage-derived exosomes in human keloid fibroblasts (HKFs) was detected using a confocal microscope. miRNA sequencing was used to explore the differentially expressed miRNAs in exosomes derived from the normoxic and hypoxic macrophage. Subsequently, the dual-luciferase reporter assay verified that phosphatase and tension homolog (PTEN) was miR-26b-5p's target. The biological function of macrophage-derived exosomes, miR-26b-5p and PTEN were detected using the CCK-8, wound-healing and Transwell assays. Western blot assay was used to confirm the miR-26b-5p's underlying mechanisms and PTEN-PI3K/AKT pathway.

Results: We demonstrated that M2-type macrophages were enriched in keloids and that hypoxia treatment could polarize macrophages toward M2-type. Compared with normoxic macrophages-derived exosomes (NMDE), HMDE promote the proliferation, migration and invasion of HKFs. A total of 38 differential miRNAs (18 upregulated and 20 downregulated) were found between the NMDE and HMDE. miR-26b-5p was enriched in HMDE, which could be transmitted to HKFs. According to the results of the functional assay, exosomal miR-26b-5p produced by macrophages facilitated HKFs' migration, invasion and proliferation via the PTEN-PI3K/AKT pathway.

Conclusions: The highly expressed miR-26b-5p in HMDE promotes the development of keloids via the PTEN-PI3K/AKT pathway.

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缺氧巨噬细胞源性外泌体miR-26b-5p靶向PTEN促进瘢痕疙瘩的发展。

背景：缺氧是瘢痕疙瘩的典型特征：缺氧是瘢痕疙瘩的典型特征。瘢痕疙瘩的发生与巨噬细胞的异常表型转变密切相关。然而，来自缺氧巨噬细胞的外泌体微RNA（miRNA）在瘢痕疙瘩中的作用仍不清楚。本研究旨在探讨缺氧巨噬细胞衍生的外泌体（HMDE）在瘢痕疙瘩发生和发展中的作用，并确定关键的miRNA：方法：通过免疫荧光检测CD206+ M2巨噬细胞在瘢痕疙瘩和正常皮肤组织中的表达。通过流式细胞术检测巨噬细胞在缺氧环境下的极化。利用共聚焦显微镜检测了巨噬细胞外泌体在人瘢痕疙瘩成纤维细胞（HKFs）中的内化情况。随后，双荧光素酶报告实验验证了磷酸酶和张力同源物（PTEN）是miR-26b-5p的靶标。利用 CCK-8、伤口愈合和 Transwell 试验检测了巨噬细胞外泌体、miR-26b-5p 和 PTEN 的生物功能。结果表明，M2型巨噬细胞中的miR-26b-5p与PTEN-PI3K/AKT通路密切相关：结果：我们证实，M2型巨噬细胞在瘢痕疙瘩中富集，缺氧处理可使巨噬细胞向M2型极化。与常氧巨噬细胞衍生的外泌体（NMDE）相比，HMDE能促进香港瘢痕疙瘩的增殖、迁移和侵袭。NMDE和HMDE共发现了38个不同的miRNAs（18个上调，20个下调），其中miR-26b-5p在HMDE中富集，可传递给HKFs。功能检测结果显示，巨噬细胞产生的外泌体miR-26b-5p通过PTEN-PI3K/AKT通路促进了HKFs的迁移、侵袭和增殖：结论：高表达的miR-26b-5p通过PTEN-PI3K/AKT途径促进瘢痕疙瘩的发展。

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来源期刊

Burns & Trauma 医学-皮肤病学

CiteScore

8.40

自引率

9.40%

发文量

186

审稿时长

6 weeks

期刊介绍： The first open access journal in the field of burns and trauma injury in the Asia-Pacific region, Burns & Trauma publishes the latest developments in basic, clinical and translational research in the field. With a special focus on prevention, clinical treatment and basic research, the journal welcomes submissions in various aspects of biomaterials, tissue engineering, stem cells, critical care, immunobiology, skin transplantation, and the prevention and regeneration of burns and trauma injuries. With an expert Editorial Board and a team of dedicated scientific editors, the journal enjoys a large readership and is supported by Southwest Hospital, which covers authors'' article processing charges.