白癜风患者病变表皮与正常表皮的比较蛋白质组学研究  

Comparative Proteomic Study on The Lesional and Non-lesional Epidermis From Vitiligo Patients

作　　者：艾力克木·吐尔逊 陈秀兰 艾尼瓦尔·塔力甫[4] 李娜[2,3] 王继峰 蔡潭溪[2,3] 郭晓静 丁翔 谢振声[3] 牛丽丽 张朦朦[3] Ghulam Abbas[2,3] 阿吉艾克拜尔·艾萨[1,2] 杨福全[2,3] Ailikemu Tuerxun;CHEN Xiu-Lan;Ainiwaer Talifu;LI Na;WANG Ji-Feng;CAI Tan-Xi;GUO Xiao-Jing;DING Xiang;XIE Zhen-Sheng;NIU Li-Li;ZHANG Meng-Meng;Ghulam Abbas;Haji Akber Aisa;YANG Fu-Quan(Key Laboratory of Plant Resources and Chemistry of Arid Zone,Xinjiang Technical Institute of Physics and Chemistry,Chinese Academy of Sciences,Urumqi 830011,China;University of Chinese Academy of Sciences,Beijing 100049,China;Laboratory of Protein and Peptide Pharmaceuticals and Laboratory of Proteomics,Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China;Hospital of Xinjiang Traditional Uyghur Medicine,Urumqi 830049,China)

机构地区：[1]中国科学院新疆理化技术研究所干旱区植物资源化学重点实验室,乌鲁木齐830011 [2]中国科学院大学,北京100049 [3]中国科学院生物物理研究所,北京100101 [4]新疆维吾尔自治区维吾尔医医院,乌鲁木齐830049

出　　处：《生物化学与生物物理进展》2023年第2期334-345,共12页Progress In Biochemistry and Biophysics

基　　金：国家重点研发计划(2020YFE0205600)资助项目。

摘　　要：目的本文通过开展白癜风患者病变表皮与正常表皮的比较蛋白质组学研究,发现和鉴定白癜风患者病变表皮与正常表皮之间的差异表达蛋白,以探讨白癜风患者表皮发生病变的分子机制。方法首先,建立和优化了表皮样品中蛋白质的最佳酶切条件。其次,采用基于串联质谱标签(TMT)标记的定量蛋白质组学技术策略开展了稳定期白癜风患者病变表皮与正常表皮的比较蛋白质组学研究,并筛选了差异表达蛋白。最后通过生物信息学分析工具及数据库(GO、KEGG、STRING、GSEA)对差异蛋白进行功能富集分析。结果优化所得到的最佳酶解条件是由Lys-C(酶∶底物,1∶100)和胰酶(酶∶底物,1∶50)组合而成的顺序酶切。比较蛋白质组学研究共鉴定4496个蛋白质,其中181个蛋白质为白癜风患者病变表皮中的差异表达蛋白。生物信息学分析表明差异表达蛋白主要与代谢、免疫、氧化还原和细胞黏附相关。其中119个上调蛋白主要参与角质化、转录、氧化应激及蛋白酶解等过程。62个下调蛋白主要参与细胞内物质运输、谷胱甘肽代谢和肌动蛋白细丝封端等过程。结论比较蛋白质组学研究揭示了白癜风患者病变表皮与正常表皮之间主要存在角质化、免疫、脂质代谢和氧化还原等方面的功能差异,发现了PRDX1、PRDX2、EEF2、ITGB1、SPTBN2、ANXA1及PFKL等蛋白质为潜在的白癜风患者病变表皮功能失调的关键蛋白质。Objective The comparative proteomic study on the paired lesional epidermis(LE) and non-lesional epidermis(NLE) from vitiligo patients to identify the differentially expressed proteins(DEPs) between LE and NLE, and to further explore the molecular mechanism of pathogenesis of vitiligo. Methods Firstly, the in solution digestion condition for proteins from epidermis were optimized to sequential tandem digestion with Lys-C and trypsin. Secondly, tandem mass tag(TMT) based quantitative proteomic strategy was performed to compare the proteome profile of the paired LE and NLE from three stable non-segmental vitiligo subjects, and differential expressed proteins(DEPs) were identified. At last, the functional enrichment analysis was performed via bioinformatics tool and database(GO, KEGG, STRING, GSEA). Results The optimal sequential tandem digestion condition was the combination of Lys-C(enzyme∶ substrate, 1∶ 100) and trypsin(enzyme∶ substrate, 1∶ 50). A total of 4 496 proteins were identified, and of which 181 were DEPs between LE and NLE from vitiligo patients. Bioinformatics analysis showed that DEPs were mainly related with metabolism, immunity, redox and cell adhesion. Among them, the 119 up-regulated proteins are mainly involved in the processes of keratinization, transcription, oxidative stress, and proteolysis. The 62 down-regulated proteins are mainly involved in intracellular transport, glutathione metabolism and actin filament capping. Conclusion The comparative proteomic study revealed that there were functional differences in keratinization, immunity, lipid metabolism and redox between LE and NLE in vitiligo patients. PRDX1, PRDX2, EEF2, ITGB1, SPTBN2, ANXA1 and PFKL were found as the key proteins to disfunction of LE.

关 键 词：白癜风 表皮蛋白组学 TMT标记定量 角质化 氧化应激 脂质代谢 

分 类 号：Q51[生物学—生物化学] Q594