蛋白组学分析寻常型银屑病患者皮损中的差异表达蛋白

Investigation of the protein expression level in lesions of patients with psoriasis vulgaris by proteomics technology

目的:通过蛋白组学技术探讨寻常型银屑病患者皮损中的差异表达蛋白。方法:纳入6例于2020年6月—9月在我院就诊的寻常型银屑病(PV)患者和6例色素痣患者作为研究对象,手术获取PV患者皮损和色素痣皮损周边正常皮肤组织。通过蛋白组学技术筛选出PV患者(n=4)皮损与正常皮肤组织(n=4)中的差异表达蛋白,并将差异表达蛋白进行GO富集和KEGG富集分析,通过蛋白互作(PPI)分析筛选出PV皮损中的核心蛋白。将核心蛋白在2例PV皮损和2例正常皮肤组织中进行Western blot验证。结果:相对于正常皮肤组织,PV皮损中发生显著上调的蛋白有291个,显著下调的蛋白有163个,涉及的生物学过程主要为中性粒细胞活化、中性粒细胞介导的免疫、TNF-α介导的信号通路、B细胞介导的免疫、上皮细胞分化及角质化为主,涉及的发病机制可能与IL-17信号通路、氧化磷酸化、PPAR信号通路等有关。PPI结果显示有12个蛋白(S100A7、S100A9、neutrophil gelatinase-associated lipocalin、cytochrome c oxidase subunit NDUFA4、fatty acid-binding protein 5、peroxisomal acyl-coenzyme A oxidase 1、platelet glycoprotein 4、cathepsin B、eosinophil cationic protein、haptoglobin、arginase-1和apoptosis-associated speck-like protein containing a CARD)可能在银屑病的发病机制中扮演重要作用。相对于正常皮肤组织,PV患者皮损中S100A7 (t=5.54)、S100A9 (t=45.23)、Cathepsin B(t=4.23)、Haptoglobin(t=4.12)和Arginase-1(t=6.65)蛋白表达水平升高,差异均具有统计学意义(均P<0.05)。结论:多种免疫信号通路及差异表达蛋白可能在PV的发病机制中扮演了重要作用,为PV的治疗靶点研究进一步提供了方向。

Objective:To investigate the differentially expressed proteins in lesions of patients with psoriasis vulgaris(PV) by proteomics technology. Methods:A total of six patients with PV and six patients with pigmented nevus presented to our hospital between June 2020 and September 2020 were enrolled. The lesions of PV and pigmented nevus-adjacent normal skin tissues were obtained by surgical removation. These differentially expressed proteins between the lesions of patients with PV(n=4) and normal skin tissues(n=4) was explored by proteomics technique. These differentially expressed proteins were further analyzed by GO and KEGG enrichment analysis. The protein protein interaction(PPI) analysis was used to screen out key proteins in the lesions of PV. Key proteins were independently verified in the lesions with PV(n=2) and normal skin tissues(n=2). Results:When compared with normal skin tissues, 291 proteins were significantly up-regulated and 163 proteins were significantly down-regulated in lesions of PV. These involved biological processes were mainly associated with neutrophil activation, neutrophil-mediated immunity, TNF-α-mediated signaling pathway, B cell-mediated immunity, epithelial cell differentiation and keratinization. The involved pathogenesis was mainly associated with IL-17 signaling pathway, oxidative phosphorylation, PPAR signal pathways and others. The PPI results showed that S100 A7, S100 A9, neutrophil gelatinase-associated lipocalin, cytochrome c oxidase subunit NDUFA4, fatty acid-binding protein 5, peroxisomal acyl-coenzyme A oxidase 1, platelet glycoprotein 4, cathepsin B, eosinophil cationic protein, haptoglobin, haptog-1 lobin, apoptosis-associated speck-like protein containing a CARD may paly the key role in the pathogenesis of PV. When compared with normal skin tissues, these expression levels of S100 A7(t=5.54, P<0.05), S100 A9(t=45.23, P<0.05), cathepsin B(t=4.23, P<0.05), haptoglobin(t=4.12, P<0.05) and arginase-1(t=6.65, P<0.05) proteins in the lesions of PV were significantly increased, and the differences were statistically significant. Conclusions:The mutiple immune-related pathways and differentially expressed proteins may paly an important role in the pathogenesis of PV, which could provide new therapeutic targets for PV.