酒精性肝病患者血浆外泌体差异蛋白的分析

目的筛选酒精性肝病(alcoholic liver disease, ALD)患者血浆外泌体中的差异蛋白,分析其功能及生物学过程,为ALD患者的治疗和诊断提供参考依据。方法选取2021年5月至2021年10月在首都医科大学附属北京佑安医院住院并确诊为ALD的患者和健康体检者各3例,超速离心分离提纯外泌体,提取蛋白,串联质谱标签(tandem mass tag, TMT)标记后,采用定量蛋白组学技术对两者血浆中的外泌体蛋白进行鉴定和定量分析,筛选出差异蛋白,并用生物信息学分析差异蛋白的功能及其参与的生物学过程。结果共鉴定到可定量蛋白质387种,以倍数上调>1.2倍或下调>1.2倍且P<0.05为标准筛选出差异蛋白27种,与健康对照组相比,ALD组上调蛋白15种、下调蛋白12种,生物信息学分析结果显示,这些蛋白主要参与了脂质的代谢、免疫反应和细胞的死亡等生物学过程,并与炎症反应、细胞的损伤和补体级联反应等信号通路密切相关。结论 TMT标记定量蛋白质组学技术筛选出的差异蛋白可能作为ALD早期诊断的血清学标志物及治疗靶点。

Analysis of differential proteins of plasma exosome in patients with alcoholic liver disease

Objective To screen the differential proteins in plasma exosome of patients with alcoholic liver disease(ALD), analyze their functions and biological processes, and provide a reference for the treatment and diagnosis of ALD patients. Methods A total of 3 ALD patients and 3 healthy examiners who were hospitalized and diagnosed in Beijing YouAn Hospital, Capital Medical University from May 2021 to Oct. 2021 were selected. Ultracentrifugation was used to separate and purify exosome, extract proteins, and tandem mass tag(TMT). After labeling, quantitative proteomics technology was used to identify and quantitatively analysis the exosome proteins in the plasma of the two, screen out the differential proteins, and use bioinformatics to analyze the functions of the differential proteins and the biological processes that they participated in. Results A total of 387 quantifiable proteins were identified, and 27 differential proteins were screened based on the criteria of up-regulation > 1.2 folds or down-regulation > 1.2 folds and P<0.05. Compared with the healthy control group, 15 up-regulated proteins and 12 down-regulated proteins in the ALD group, bioinformatics analysis results showed that these proteins were mainly involved in lipid metabolism, immune response and cell death and other biological processes, and were closely related to signaling pathways such as inflammation, cell damage, and complement cascade. Conclusion The differential proteins screened by TMT-labeled quantitative proteomics technology may be used as serological markers and therapeutic targets for early diagnosis of ALD.