摘要
目的:探讨基于定量蛋白质组学串联质谱标签(TMT)技术筛选糖尿病足皮肤抗菌肽的可行性和价值。方法:收集2017年4月至2018年12月在东南大学附属中大医院行清创手术的糖尿病足患者的下肢溃疡周围皮肤全层和同期行其他手术非糖尿病患者的下肢皮肤全层标本各3例患者标本。利用TMT技术分别对上述标本进行蛋白组学分析,筛选出发生显著变化的差异蛋白。搜索UniProt-GOA数据库对鉴定到的差异蛋白生物学作用阐释。以鉴定到的蛋白为背景,利用Fisher精确检验法评价差异表达蛋白。利用InterPro数据库分析差异表达蛋白功能结构域的富集情况。结果:共发现133个差异蛋白,其中表达上调和下调的蛋白分别为101个和32个。基因本体(GO)富集分析表明,在蛋白生物学过程中最显著富集的过程为对细菌的防御过程,其次为抗菌体液反应过程。共有19个蛋白涉及对细菌的防御反应过程,其中18个蛋白发生了上调,1个蛋白发生了下调。差异蛋白上调倍数达4倍以上的共有8个,分别为蛋白为溶菌酶C、中性粒细胞弹性蛋白酶、杀菌渗透性增加蛋白、乳铁蛋白、膜相关磷脂酶A2、肽聚糖识别蛋白1、免疫球蛋白J链和蛋白S100-A12。共有8个蛋白涉及抗菌体液反应过程,其中7个蛋白与对细菌的防御反应中涉及的蛋白重叠。通过蛋白互作分析显示,差异表达的抗菌反应蛋白形成了一个包含14个节点和34条边的复杂调控网络,平均节点度为4.86,聚类系数为0.652。结论:通过TMT标记蛋白组学技术发现,参与对细菌的防御反应及抗菌体液免疫反应的差异蛋白,如溶菌酶C、中性粒细胞弹性蛋白酶、杀菌渗透性增加蛋白、乳铁蛋白、膜相关磷脂酶A2、肽聚糖识别蛋白1、免疫球蛋白J链和蛋白S100-A12,有望成为治疗糖尿病足感染的新靶点。
Objective To explore the feasibility and utility of quantitative proteomics tandem mass tag(TMT)in screening antimicrobial peptides from diabetic foot skin.Methods During April 2017 to December 2018,3 samples from the skin around the ulcer of the lower extremity in patients with diabetic foot who underwent debridement,and another 3 from the skin around the ulcer of the lower extremity in non-diabetic patients who received other operations in Zhongda Hospital,Southeast University,were collected.TMT was used to analyze the proteomics of these samples,and the significantly changed proteins were screened.The biological functions of the identified differential proteins were interpreted based on UniProt-GOA database.Using the identified proteins as the background,Fisher exact test was used to evaluate the function of differentially expressed proteins.The magnitude of the enrichment of functional domains of differentially expressed proteins was analyzed using InterPro database.Results A total of 133 differentially expressed proteins were identified,of which 101 and 32 were up-regulated and down-regulated,respectively.Gene ontology(GO)enrichment analysis showed that the most significant enrichment process in protein biology was the defense process against bacteria,followed by the humoral response process against bacteria.A total of 19 proteins involved in the defense response process to bacteria,of which 18 proteins were up-regulated and 1 protein was down-regulated.Eight proteins were up-regulated more than four-fold,which included Lysozyme C,neutrophil elastase,bactericidal permeability increasing protein,lactoferrin,membrane-associated phospholipase A2,peptidoglycan recognition protein 1,immune globulin J chain,and protein S100-A12.A total of 8 proteins were involved in the process of antimicrobial humoral response,of which 7 proteins overlapped with proteins involved in the defense response to bacteria.Protein interaction analysis showed that the differentially expressed antimicrobial proteins formed a complex regula
作者
刘德林
殷汉
杨兵全
嵇振岭
Liu Delin;Yin Han;Yang Bingquan;Ji Zhenling(Department of Endocrinology,Zhongda Hospital,Southeast University,Nanjing 210009,China;Department of General Surgery,Zhongda Hospital,Southeast University,Nanjing 210009,China)
出处
《中华糖尿病杂志》
CAS
CSCD
北大核心
2020年第11期923-928,共6页
CHINESE JOURNAL OF DIABETES MELLITUS
基金
中央高校基本科研业务费专项(3290002043C2)
中国微循环学会重点项目(TW2018C001)。
关键词
糖尿病足
蛋白组学
抗菌肽
串联质谱标签
Diabetic foot
Proteomic
Antimicrobial peptides
Tandem mass tag
