中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (3): 435-440.doi: 10.3969/j.issn.2095-4344.0594

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

乌司他丁干预减体积肝移植模型大鼠的肝脏代谢

高红强,刘  静,李志强,王海雷,赵雄齐,张升宁,冉江华,李  立   

  1. (昆明医科大学附属甘美医院、昆明市第一人民医院肝胆胰外科,云南省昆明市   650011)
  • 收稿日期:2018-08-07
  • 通讯作者: 刘静,博士,副主任医师,昆明医科大学附属甘美医院、昆明市第一人民医院肝胆胰外科,云南省昆明市 650011
  • 作者简介:高红强,男,1981年生,汉族,云南省曲靖市人,2016年昆明医科大学外科学专业毕业,博士,主治医师,主要从事肝胆胰外科及器官移植方面的研究。
  • 基金资助:

    云南省应用基础研究计划(2013FZ218),项目负责人:刘静;云南省卫生科技计划(2014NS199),项目负责人:李立

Ulinastatin improves rat liver metabolism after reduced-size liver transplantation 

Gao Hongqiang, Liu Jing, Li Zhiqiang, Wang Hailei, Zhao Xiongqi, Zhang Shengning, Ran Jianghua, Li Li   

  1. (Department of Hepato-Biliary-Pancreatic Surgery, Calmette Hospital & the First Hospital of Kunming, Kunming Medical University, Kunming 650011, Yunnan Province, China)
  • Received:2018-08-07
  • Contact: Liu Jing, MD, Associate chief physician, Department of Hepato-Biliary-Pancreatic Surgery, Calmette Hospital & the First Hospital of Kunming, Kunming Medical University, Kunming 650011, Yunnan Province, China
  • About author:Gao Hongqiang, MD, Attending physician, Department of Hepato-Biliary-Pancreatic Surgery, Calmette Hospital & the First Hospital of Kunming, Kunming Medical University, Kunming 650011, Yunnan Province, China
  • Supported by:

    the Basic and Applied Research Project of Yunnan Province, No. 2013FZ218 (to LJ); the Health and Scientific Technology Program of Yunnan Province, No. 2014NS199 (to LL)

摘要:

文章快速阅读:

文题释义:
基因表达谱芯片:基因表达谱芯片是采用 cDNA 或寡核苷酸片段作探针,固化在芯片上;将实验样品与对照样品的 mRNA 以两种不同的荧光分子进行标记,同时与芯片进行杂交,分析两种样品与探针杂交的荧光强度的比值,来检测基因表达水平的变化。
蛋白质质谱分析:蛋白质经过蛋白酶酶切后成肽段混合物,经质量分析器得到蛋白质的一级质谱峰图。离子选择装置自动选取强度较大肽段离子进行二级质谱分析,输出选取肽段的二级质谱峰图,通过和已知蛋白质的两级质谱峰图进行比对而鉴定蛋白质。
摘要
背景
:乌司他丁在肝切除、肝移植后抗炎、脏器保护、改善微循环等方面的作用已经有大量的研究,然而其作用的microRNA调控机制尚未见报道。
目的:观察模型大鼠减体积肝移植后乌司他丁干预microRNA及蛋白组学的变化,并在差异表达的microRNAs和蛋白中预测microRNA对其靶向蛋白的调控,为乌司他丁的临床应用提供更深入的理论依据。
方法:以Kamada的双袖套法为基础建立40%减体积肝移植模型大鼠,实验分2组:实验组在肝移植后0,12,24,36,48 h经腹腔注入乌司他丁(100 U/g);对照组在同样时间点注入生理盐水2 mL。在移植后24,48 h取2组大鼠肝脏分别行microRNA芯片检查及蛋白质质谱分析。将二者的结果输入mirTarBase软件进行靶基因预测。
结果与结论:①与对照组相比,实验组表达差异超过2倍的miRNAs有19个,蛋白丰度表达差异超过1.5倍的蛋白有17个,靶基因预测发现一组具有强力证据推荐的microRNA目标蛋白的调控通道:rno-miR-181a-5p和Gpx1;②结果表明,模型大鼠减体积肝移植后,乌司他丁的使用,改变了rno-miR-181a-5p的表达,rno-miR-181a-5p通过调控Gpx1表达来改善模型大鼠减体积肝移植术后肝脏的代谢,这可能是乌司他丁作用机制之一。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0002-6765-1030(高红强)

关键词: 乌司他丁, microRNAs芯片, 蛋白质质谱,miR-181-a-5p, 大鼠减体积肝移植, 缺血再灌注损伤, 组织构建

Abstract:

BACKGROUND: Effects of ulinastatin in anti-inflammatory, viscera protection and improving microcirculation after liver resection or liver transplantation have been widely researched. However, the role of microRNA regulation mechanism of ulinastatin has not yet been reported.
OBJECTIVE: To discover the variation of microRNA expression profile and the change of proteomics in rats after reduced-size liver transplantation using ulinastatin, and to investigate if there is a lien between differentially expressed miroRNAs and proteins, so as to provide an in-depth theoretical evidence for clinical application of ulinastatin.
METHODS: Rat models of reduced-size liver transplantation were established using the Kamada double cuff method and then divided into two groups. Rats in the experimental and control groups were given intraperitoneal injection of ulinastatin (100 U/g) and normal saline (2 mL) respectively at 0, 12, 24, 36, and 48 hours after surgery. miroRNA chip and proteomic analysis were performed at 24 and 48 hours after transplantation. Then results were then imported into mirTarBase software for target prediction.
RESULTS AND CONCLUSION: Compared with the control group, in the experimental group, there were 19 differentially expressed (> two-fold) proteins and 17 differentially expressed (> 1.5 fold) proteins in rats. A miRNA regulation channel: miR-181-a-5p and Gpx1 were found. To conclude, after reduced-size liver transplantation in rats, use of ulinastatin alters miR-181-a-5p expression, which improves liver metabolism through regulating Gpx1 expression. This may be one mechanism of action of ulinastatin.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: MicroRNAs, Mass Spectrometry, Liver Transplantation, Reperfusion Injury, Tissue Engineering

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