溶菌酶与循环RNA、细胞外囊泡及慢性应激的关联。
Lysozyme association with circulating RNA, extracellular vesicles, and chronic stress.
作者信息
Abey Sarah K, Yuana Yuana, Joseph Paule V, Kenea Natnael D, Fourie Nicolaas H, Sherwin LeeAnne B, Gonye Gregory E, Smyser Paul A, Stempinski Erin S, Boulineaux Christina M, Weaver Kristen R, Bleck Christopher K E, Henderson Wendy A
机构信息
Digestive Disorders Unit, Division of Intramural Research, National Institutes of Health, Department of Health and Human Services, Bethesda, MD, USA.
Image Sciences Institute, Division of Imaging, University Medical Centre Utrecht, Netherlands.
出版信息
BBA Clin. 2016 Dec 20;7:23-35. doi: 10.1016/j.bbacli.2016.12.003. eCollection 2017 Jun.
BACKGROUND
Stress has demonstrated effects on inflammation though underlying cell-cell communication mechanisms remain unclear. We hypothesize that circulating RNAs and extracellular vesicles (EVs) in patients with chronic stress contain signals with functional roles in cell repair.
METHODS
Blood transcriptome from patients with Irritable Bowel Syndrome versus controls were compared to identify signaling pathways and effectors. Plasma EVs were isolated (size-exclusion chromatography) and characterized for effectors' presence (immunogold labelling-electron microscopy). Based on transcriptome pathways and EV-labelling, lysozyme's effects on cell migration were tested in human colon epithelial CRL-1790 cells and compared to the effects of CXCL12, a migration inducer (wound assay). The effect of lysozyme on immune-linked mRNA and protein levels in cells which survived following serum starvation and scratch wound were investigated (NanoString).
RESULTS
Blood transcriptomes revealed pyridoxal 5'phosphate salvage, pyrimidine ribonucleotides salvage pathways, atherosclerosis, and cell movement signaling with membrane CD9 and extracellular lysozyme as effectors. Plasma EVs showed labelling with CD9, mucins, and lysozyme. This is the first identification of lysozyme on plasma EVs. In CRL-1790 cells, lysozyme induced migration and repaired scratch wound as well as CXCL12. Immune mRNA and protein expressions were altered in cells which survived following serum starvation and scratch wound, with or without lysozyme in serum-free media post-wounding: CD9, IL8, IL6 mRNAs and CD9, NT5E, PD-L1 proteins.
CONCLUSIONS
Repair and inflammatory signals are identified in plasma EVs and circulating RNAs in chronic stress. Registered clinicaltrials.gov #NCT00824941.
GENERAL SIGNIFICANCE
This study highlights the role of circulating RNAs and EVs in stress.
背景
压力已被证明对炎症有影响，但其潜在的细胞间通讯机制仍不清楚。我们假设慢性应激患者循环中的RNA和细胞外囊泡（EVs）含有在细胞修复中起作用的信号。
方法
比较肠易激综合征患者与对照组的血液转录组，以确定信号通路和效应物。分离血浆EVs（尺寸排阻色谱法），并通过免疫金标记电子显微镜鉴定效应物的存在。基于转录组通路和EV标记，在人结肠上皮CRL-1790细胞中测试溶菌酶对细胞迁移的影响，并与迁移诱导剂CXCL12的作用进行比较（伤口试验）。研究了溶菌酶对血清饥饿和划痕伤口后存活细胞中免疫相关mRNA和蛋白质水平的影响（NanoString）。
结果
血液转录组显示以膜CD9和细胞外溶菌酶为效应物的磷酸吡哆醛5'-磷酸补救、嘧啶核糖核苷酸补救途径、动脉粥样硬化和细胞运动信号。血浆EVs显示有CD9、粘蛋白和溶菌酶的标记。这是首次在血浆EVs上鉴定出溶菌酶。在CRL-1790细胞中，溶菌酶诱导迁移并修复划痕伤口，其效果与CXCL12相同。在血清饥饿和划痕伤口后存活的细胞中，免疫mRNA和蛋白质表达发生改变，在伤口后无血清培养基中添加或不添加溶菌酶：CD9、IL8、IL6 mRNA以及CD9、NT5E、PD-L1蛋白质。
结论
在慢性应激的血浆EVs和循环RNA中鉴定出修复和炎症信号。美国国立医学图书馆临床试验注册中心编号#NCT00824941。
普遍意义
本研究强调了循环RNA和EVs在应激中的作用。
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