School of Clinical Medicine, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi 330004, China.
Department of Integrated Traditional Chinese and Western Medicine, Dermatology Hospital of Jiangxi, Nanchang, Jiangxi 330001, China.
Oxid Med Cell Longev. 2022 Oct 12;2022:6264474. doi: 10.1155/2022/6264474. eCollection 2022.
NHEKs, HaCaT cells, and HEK 293T cells were treated with IL-17A. CCK-8 assays were performed to detect cell activity, and immunofluorescence staining and Western blotting were performed to detect the protein expression of STAT3. After isolation of exosomes via ultracentrifugation, the contents of miR-124-3p and oxidative stress markers such as superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) in keratinocytes were measured. Subsequently, transcriptomic analysis was performed using RNA-seq. Data were analysed by using the "edgeR" package within R. After verifying the abnormally expressed genes stimulated by IL-17A, a dual luciferase reporter assay was used to determine the interaction between miR-124-3p and STAT3. Finally, BALB/c mice were used to establish a psoriasis model for analysis. The effect of elevated miR-124-3p on the psoriasis mouse model was determined by exosomal delivery of miR-124-3p.
IL-17 intervention enhanced the cell activity of keratinocytes ( < 0.05). miR-124-3p was identified by RNA-seq as one of the differentially expressed miRNAs stimulated by IL-17A. miR-124-3p overexpression induced decreased STAT3 and MDA levels, increased SOD and GSH-Px levels in keratinocytes, and alleviated emergency responses of sclerosis damage ( < 0.05). The dual luciferase reporter assay results confirmed that STAT3 was regulated by miR-124-3p in a targeted manner ( < 0.05). Finally, miR-124-3p delivered by exosomes effectively alleviated the pathological manifestations and oxidative stress responses of psoriatic mice.
miR-124-3p regulates keratinocyte activity via STAT3 in response to IL-17A stimulation. The ectopic expression of miR-124-3p in psoriatic skin reduces IL-17A-induced inflammation and inhibits the STAT3 pathway, thus alleviating the symptoms of psoriasis. The findings of this study suggest that exosomes can be used to therapeutically deliver miR-124-3p to keratinocytes and psoriatic lesions, which may provide novel insight for psoriasis treatment.
采用白细胞介素 17A（IL-17A）处理正常人表皮角质形成细胞（NHEKs）、HaCaT 细胞和人胚肾 293T 细胞。通过 CCK-8 检测试剂盒检测细胞活性，通过免疫荧光染色和 Western blot 检测 STAT3 蛋白的表达。通过超速离心分离角质形成细胞中的外泌体，测量外泌体中 miR-124-3p 的含量和氧化应激标志物（超氧化物歧化酶（SOD）、丙二醛（MDA）和谷胱甘肽过氧化物酶（GSH-Px））的含量。然后，使用 RNA-seq 进行转录组分析。使用 R 中的“edgeR”包分析数据。验证了 IL-17A 刺激下异常表达的基因后，使用双荧光素酶报告基因检测 miR-124-3p 与 STAT3 之间的相互作用。最后，使用 BALB/c 小鼠建立银屑病模型进行分析。通过外泌体递送 miR-124-3p 来确定升高的 miR-124-3p 对银屑病小鼠模型的影响。
IL-17 干预增强了角质形成细胞的细胞活性（<0.05）。通过 RNA-seq 鉴定到 miR-124-3p 是 IL-17A 刺激下差异表达的 miRNAs 之一。miR-124-3p 过表达诱导角质形成细胞中 STAT3 和 MDA 水平降低，SOD 和 GSH-Px 水平升高，并减轻硬化损伤的应急反应（<0.05）。双荧光素酶报告基因检测结果证实 STAT3 受 miR-124-3p 靶向调控（<0.05）。最后，外泌体递送的 miR-124-3p 有效缓解了银屑病小鼠的病理表现和氧化应激反应。
miR-124-3p 通过 STAT3 调节角质形成细胞对白细胞介素 17A 刺激的活性。在银屑病皮肤中异位表达 miR-124-3p 可减少 IL-17A 诱导的炎症，并抑制 STAT3 通路，从而缓解银屑病的症状。本研究结果表明，外泌体可以用于治疗性递送至角质形成细胞和银屑病病变部位的 miR-124-3p，这可能为银屑病治疗提供新的思路。
Zotero 插件
