Yan Jianjun, Zhen Yunyue, Wang Ruijie, Li Xueqing, Huang Shan, Zhong Hua, Wen He, Sun Qing
Department of Dermatology, Qilu Hospital of Shandong University, Jinan, China.
Laboratory of Basic Medical Science, Qilu Hospital of Shandong University, Jinan, China.
Front Genet. 2022 Aug 10;13:891465. doi: 10.3389/fgene.2022.891465. eCollection 2022.
The role of serum extracellular vesicles (EVs) is less known in psoriasis. To explore the transcriptomic profile of serum EVs and the potential biomarkers in psoriasis. EVs were isolated by differential ultracentrifugation and identified by transmission electron microscope. The diameters of EVs were detected using nanoparticle tracking analysis. Serum EVs-keratinocyte interaction was observed through confocal fluorescence microscopy. miRNA microarray and mRNA microarray were performed in serum EVs ( = 4) and skin lesions ( = 3), respectively. Quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and fluorescence hybridization were used to detect the expression of miRNAs in serum EVs and skin lesions ( = 15). Bioinformatics analysis was performed to predict the potential target genes and functions of miR-1305 and miR-6785-5p. Western blot, CCK-8 and enzyme-linked immunosorbent assay (ELISA) were used to detect the EVs' biomarkers, keratinocytes proliferation and cytokines secretion. A total of 16 miRNAs and 1,725 mRNAs were significantly dysregulated in serum EVs and skin lesions, respectively. miR-1305 was down-regulated and miR-6785-5p was upregulated in both serum EVs and skin lesions. Serum EVs could be taken up by keratinocytes. miR-1305 was downregulated and miR-6785-5p were upregulated in keratinocytes after co-cultured with psoriasis serum EVs compared with controls. Psoriasis serum EVs promoted keratinocyte proliferation and the secretion of CCL20 and IL-8. Serum EVs miR-1305 and miR-6785-5p were associated with disease severity. Serum EVs might be involved in the activation of keratinocytes through loaded miRNAs in psoriasis. Serum EVs miR-1305 and miR-6785-5p may be associated with psoriasis.
血清细胞外囊泡(EVs)在银屑病中的作用鲜为人知。为了探索血清EVs的转录组图谱以及银屑病中的潜在生物标志物。通过差速超速离心法分离EVs,并通过透射电子显微镜进行鉴定。使用纳米颗粒跟踪分析检测EVs的直径。通过共聚焦荧光显微镜观察血清EVs与角质形成细胞的相互作用。分别对血清EVs(n = 4)和皮肤病变组织(n = 3)进行miRNA微阵列和mRNA微阵列检测。采用定量逆转录聚合酶链反应(qRT-PCR)和荧光原位杂交技术检测血清EVs和皮肤病变组织(n = 15)中miRNAs的表达。进行生物信息学分析以预测miR-1305和miR-6785-5p的潜在靶基因和功能。采用蛋白质免疫印迹法、CCK-8法和酶联免疫吸附测定(ELISA)法检测EVs的生物标志物、角质形成细胞增殖和细胞因子分泌。血清EVs和皮肤病变组织中分别共有16种miRNAs和1725种mRNAs显著失调。血清EVs和皮肤病变组织中miR-1305均下调,miR-6785-5p均上调。血清EVs可被角质形成细胞摄取。与对照组相比,与银屑病血清EVs共培养后的角质形成细胞中miR-1305下调,miR-6785-5p上调。银屑病血清EVs促进角质形成细胞增殖以及CCL20和IL-8的分泌。血清EVs中的miR-1305和miR-6785-5p与疾病严重程度相关。血清EVs可能通过在银屑病中负载miRNAs参与角质形成细胞的激活。血清EVs中的miR-1305和miR-6785-5p可能与银屑病相关。