Department of Dermatology, The Second Hospital of Jilin University, Changchun, China.
Department of Thoracic Surgery, China-Japan Union Hospital of Jilin University, Changchun, China.
J Cell Mol Med. 2021 May;25(10):4551-4561. doi: 10.1111/jcmm.16037. Epub 2021 Mar 18.
Intriguingly, microRNAs (miRs) transferred as cargo in extracellular vesicles (EVs) can modulate wound healing through their regulation of fibroblast functions. In this study, we investigated the effects of miR-106b transfer via EVs derived from human umbilical vein endothelial cells (HUVECs) on skin wound healing. Dual-luciferase reporter gene assay identified that miR-106b could target and inhibit JMJD3. RT-qPCR analysis showed EVs isolated from HUVECs had enriched expression of miR-106b. LL29 fibroblast cells and HaCaT keratinocytes were co-cultured with HUVEC-derived EVs, in which miR-106b had been up-regulated or down-regulated by its mimic or inhibitor. The co-culture with HUVEC-derived EVs increased miR-106b expression, and reduced the viability and adhesion of LL29 and HaCaT cells, whereas the inhibition of miR-106b in HUVEC-derived EVs enhanced the viability and adhesion of LL29 and HaCaT cells through up-regulation of JMJD3. Next, we showed that JMJD3 overexpression enhanced LL29 and HaCaT cell viability and adhesion through elevating RIPK3, which induced the phosphorylation of AKT during the wound-healing process. We next developed a mouse skin wound model to investigate the actions of miR-106b in vivo after 14 days. The delivery of miR-106b via HUVEC-derived EVs delayed wound healing through suppression of collagen I content and angiogenesis, but had no effects on pro-inflammatory cytokines. In conclusion, miR-106b from HUVEC-derived EVs inhibits JMJD3 and RIPK3, leading to the inhibition of skin wound healing, thus constituting a new therapeutic target.
有趣的是，作为细胞外囊泡 (EVs) 货物转移的 microRNAs (miRs) 可以通过调节成纤维细胞功能来调节伤口愈合。在这项研究中，我们研究了源自人脐静脉内皮细胞 (HUVECs) 的 EVs 中转录的 miR-106b 对皮肤伤口愈合的影响。双荧光素酶报告基因检测鉴定出 miR-106b 可以靶向并抑制 JMJD3。RT-qPCR 分析显示，从 HUVECs 分离的 EVs 富含 miR-106b 的表达。将 LL29 成纤维细胞和 HaCaT 角质形成细胞与 HUVEC 衍生的 EV 共培养，其中 miR-106b 通过其模拟物或抑制剂上调或下调。与 HUVEC 衍生的 EV 共培养增加了 miR-106b 的表达，降低了 LL29 和 HaCaT 细胞的活力和粘附，而 HUVEC 衍生的 EV 中 miR-106b 的抑制通过上调 JMJD3 增强了 LL29 和 HaCaT 细胞的活力和粘附。接下来，我们表明 JMJD3 过表达通过升高 RIPK3 增强了 LL29 和 HaCaT 细胞的活力和粘附，从而在伤口愈合过程中诱导 AKT 的磷酸化。我们接下来开发了一种小鼠皮肤伤口模型，以在 14 天后研究体内 miR-106b 的作用。通过 HUVEC 衍生的 EVs 递送 miR-106b 通过抑制胶原 I 含量和血管生成来延迟伤口愈合，但对促炎细胞因子没有影响。总之，源自 HUVEC 的 EVs 的 miR-106b 抑制 JMJD3 和 RIPK3，导致皮肤伤口愈合受到抑制，从而构成新的治疗靶标。
