文章摘要
邵慧静,王天俊,张嘉嘉,杨娜娜,孙丽洲.子痫前期胎盘样本的多肽组学分析及差异多肽功能的初步探究[J].南京医科大学学报,2021,(8):1151~1159
子痫前期胎盘样本的多肽组学分析及差异多肽功能的初步探究
Comparative peptidome analysis of preeclamptic placenta and functional study of the differential peptides
投稿时间:2021-01-19  
DOI:doi:10.7655/NYDXBNS20210807
中文关键词: 子痫前期  多肽组学  内源性多肽  滋养细胞
英文关键词: preeclampsia  peptidomics  endogenous peptide  trophoblast cell
基金项目:国家重点研发计划(2018YFC1002205);国家自然科学基金面上项目(81771603);江苏省妇幼健康科学研究项目(FRC201755)
作者单位
邵慧静 南京医科大学第一附属医院产科江苏 南京 210029 
王天俊 南京医科大学第一附属医院产科江苏 南京 210029 
张嘉嘉 南京医科大学第一附属医院产科江苏 南京 210029 
杨娜娜 南京医科大学第一附属医院产科江苏 南京 210029 
孙丽洲 南京医科大学第一附属医院产科江苏 南京 210029 
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中文摘要:
      目的:通过液相色谱?串联质谱(LC?MS/MS)探索子痫前期胎盘样本中的多肽表达,并对其中差异多肽的体外功能进行初步探究。方法:收集2019年5—9月在南京医科大学第一附属医院产科行计划性剖宫产手术的4例正常足月孕妇胎盘和4例子痫前期孕妇胎盘组织。利用超滤法、LC?MS/MS检测样本中多肽的表达水平差异。通过CCK?8法,克隆形成实验以及Transwell实验初步研究差异多肽405SPLFMGKVVNPTQK418对滋养细胞(HTR?8/SVneo)增殖、迁移能力的影响。脐静脉内皮细胞(HUVECs)成管实验初步研究该多肽对HUVECs细胞血管形成能力的影响。使用qPCR和Western blot检测多肽处理组与对照组中的金属蛋白质酶MMP2以及金属蛋白酶组织抑制剂(TIMP1)的表达。结果:共鉴定到3 582条多肽片段,48条差异表达多肽。体外功能实验表明,差异多肽405SPLFMGKVVNPTQK418能够促进滋养细胞增殖迁移以及内皮细胞的管状形成能力。多肽处理后MMP2表达明显上调,TIMP1的表达明显下调。结论:子痫前期胎盘与正常胎盘间存在差异多肽,且差异多肽405SPLFMGKVVNPTQK418能够促进滋养细胞以及内皮细胞的功能,为子痫前期发病机制研究以及治疗提供了多肽组学的支持。
英文摘要:
      Objective:This study aims to identify the peptide with different expression in preeclamptic placenta by liquid chromatography?tandem mass spectrometry(LC?MS/MS),and to study the bio?active function of the differentially expressed peptides. Methods:Placental tissues were collected from 4 normal pregnant women and 4 pregnant women with preeclampsia. All participants underwent planned cesarean section in the Department of Obstetrics,the First Affiliated Hospital of Nanjing Medical University from May 2019 to September 2019. The effects of the differential peptide 405SPLFMGKVVNPTQK418 on the proliferation and migration of trophoblast cells(HTR?8/SVneo) were studied by CCK?8 assay,clone formation assay and transwell assay. And the matrigel assay was used to study the effect of this peptide on the angiogenesis ability of human umbilical vein endothelial cells(HUVECs). Western blot was used to detect the metalloproteinase 2 (MMP2) and metalloproteinase tissue inhibitor 1 (TIMP1) in peptide?treated group and control group. Results:A total of 3 582 peptides were screened out and 48 differentially expressed peptides were identified. In vitro functional assays showed that the differential peptide 405SPLFMGKVVNPTQK418 could promote the proliferation and migration of trophoblast cells and the tube formation ability of endothelial cells. After peptide treatment,the expression of MMP2 was up?regulated and the expression of TIMP1 was down?regulated. Conclusion:A series of differentially expressed peptides were detected in preeclamptic placenta,and the differential peptide 405SPLFMGKVVNPTQK418 exhibited a promotive role in the behaviour of trophoblast cells and endothelial cells,which provides support for the study of pathogenesis and treatment of preeclampsia.
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