iTRAQ-Based Proteomic Analysis of Neonatal Kidney from Offspring of Protein Restricted Rats Reveals Abnormalities in Intraflagellar Transport Proteins
iTRAQ-Based Proteomic Analysis of Neonatal Kidney from Offspring of Protein Restricted Rats Reveals Abnormalities in Intraflagellar Transport Proteins
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基于 iTRAQ 的蛋白质组学分析，来自蛋白质限制性大鼠后代的新生肾脏，揭示了鞭毛内转运蛋白的异常
DOI：
10.1159/000484626
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Liu, Caixia
中科院分区：
文献类型：
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作者：
Liu, Xiaomei;Wang, Jun;Liu, Caixia
背景资料：众所周知，宫内不良事件可损害胎儿发育，并导致成年后肾损伤和高血压的发生。我们以前曾报道过母体蛋白质营养不良引起的胎儿宫内生长受限（IUGR）后代的肾脏指数、肾小球数量和肾小球滤过率降低。为了探讨胎儿生长受损与肾脏疾病的分子机制，我们研究了IUGR新生儿肾脏中差异表达的蛋白质（DEPs）的同量异位素标签的相对和绝对定量（iTRAQ）分析。方法：采用母体蛋白质营养不良诱发IUGR。收集新生儿肾脏;提取蛋白质;在iTRAQ标记前合并，并进行质谱分析。然后通过定量PCR（qPCR）、免疫组织化学（IHC）和/或蛋白质印迹分析评估单个标本中的五种代表性蛋白质，进一步确认质谱分析结果。结果：共检测到367个DEPs，其中263个表达上调，104个表达下调。IUGR肾和对照肾之间的阈值变化为1.2倍，P值≤ 0.05。进一步的生物信息学分析表明，这些蛋白在氧化磷酸化、嘌呤代谢、嘧啶代谢、RNA小体、剪接体组装和鞭毛内转运（IFT）等过程中发挥重要作用。经蛋白质印迹、免疫组化和Q-PCR证实，IFT家族蛋白（IFT 80、88、144）和PKD 2在IUGR肾脏中上调。表观遗传调节因子SET和MYND结构域3（SMYD 3），组蛋白赖氨酸N-甲基转移酶，和H3 K4 me 3水平也显着增强IUGR新生儿肾脏。结论：新生儿肾脏蛋白质组的首次全面分析揭示了肾脏发育的新见解，并可能对鉴定成人疾病发育起源中涉及的病理机制做出有价值的贡献。
Background: It is well recognized that adverse events in utero can impair fetal development and lead to the development of kidney injury and hypertension in adulthood. We previously reported a lower kidney index, glomeruli number, and decreased glomerular filtration rate in intrauterine growth restriction (IUGR) offspring induced by maternal protein malnutrition. To explore the molecular mechanisms linking impaired fetal growth to renal diseases, we investigated differentially expressed proteins (DEPs) in the IUGR neonatal kidneys by isobaric tags for relative and absolute quantitation (iTRAQ) analysis. Methods: We induced IUGR through maternal protein malnutrition. Neonatal kidneys were collected; the protein was extracted; pooled before iTRAQ labeling, and subjected to mass spectrometric analysis. Mass spectrometry results were then further confirmed by assessing five representative proteins in individual specimens with quantitative PCR (qPCR), immunohistochemical (IHC) and / or western blot analysis. Results: A total of 367DEPs (263 up-regulated, 104 down-regulated.) with a threshold of a 1.2-fold change and a P value ≤ 0.05 between IUGR kidneys and control kidneys were identified. Further bioinformatics analysis revealed that these proteins play important roles in oxidative phosphorylation, purine metabolism, pyrimidine metabolism, RNA small body, spliceosome assembly and intraflagellar transport (IFT). IFT family proteins (IFT80, 88,144) and PKD2 were shown to be up-regulated in IUGR kidneys, confirmed by western blotting, IHC and Q-PCR. Epigenetic modulating factors SET and MYND domain containing 3 (SMYD3), a histone-lysine N-methyltransferase, and H3K4me3 level were also remarkably enhanced in IUGR neonatal kidneys. Conclusions: This first comprehensive analysis of the neonatal kidney proteome reveals new insights in nephridial development, and may make a valuable contribution towards the identification of the pathological mechanisms involved in the developmental origins of adult disease.