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鼠肝硫酸乙酰肝素生化结构的试验分析
Experimental analysis of the biochemical structure of mouse liver heparin sulfate
【摘要】 目的评估一种硫酸乙酰肝素的生化学分析方法,并对小鼠肝脏硫酸乙酰肝素的二糖组成及分子量进行生化分析。方法充分利用硫酸乙酰肝素的物理化学特点,进行提取及纯化。用酶降解及2-AB标记后,行高效液相色谱分析。所得色谱与二糖标准色谱比较得出样品的二糖组成,与分子量色谱比较得出样品的分子量。结果纯化提取的小鼠肝脏糖胺聚糖经肝素酶消化后,可获得相近的二糖含量及二糖组成。0S二糖,含量最高为(7.01±0.81)pmol/mg干重,占二糖总量的58.27%,其次为NS二糖,含量为(3.47±0.99)pmol/mg,占28.27%,再次依次为6S二糖[(0.85±0.99)pmol/mg,6.87%],Tris二糖[(0.50±0.33)pml/mg,3.85%],S1二糖(0.21pmol/mg,1.65%)和S2二糖(0.14pmol/mg,1.1%)。硫酸乙酰肝素二糖总量为(12.17±2.44)pmol/mg。硫酸乙酰肝素的分子量大小约为28kD。结论采用该文提取纯化方法的硫酸乙酰肝素纯度高,得出的小鼠肝脏硫酸乙酰肝素二糖组成及分子量可信度高,适于用来分析各种肝脏疾病时硫酸乙酰肝素的生化改变。
【Abstract】 [Objective] To evaluate a biochemical analysis method for heparin sulfate and to experimentally analyze the disaccharide composition and chain length of mouse liver heparin sulfate. [Methods] Extraction and purification of the heparan sulfate were adapted by taking the advantage of its physical and chemical properties. The purified samples were labeled with 2-Aminobenzomide (2-AB) followed by high performance liquid chromatography (HPLC) analysis. Resultant chromatography was compared with the disaccharides standards to get the disaccharide composition and amount compared with the molecular standards to know the molecular weight. [Results] Similar disaccharide composition and disaccharide amount were obtained by digesting and analyzing the glycansaminoglycans purified from the mouse liver. The 0S disaccharide was (7.01±0.81) pmol/mgdry weight,around 58.27% of the total disaccharides,next was NS disaccharide, (3.47±0.99) pmol/mg, 28.27% of the total disaccharides,following was 6S disaccharide[(0.85±0.99) pmol/mg, 6.87%], Tris disaccharide[(0.50±0.33) pml/mg, 3.85%], S1 disaccharide(0.21 pmol/mg, 1.65%) and S2 disaccharide (0.14 pmol/mg, 1.1%). The total disaccharide amount of heparin sulfate was (12.17±2.44) pmol/mg. The molecular weight of heparin sulfate chain was around 28 kD. [Conclusion] High purity heparin sulfate can be obtained by using the extraction and purification methods introduced in this article. The reproducible heparan sulfate disaccharide composition and chain length were obtained by analyzing the purified samples indicating the methods are suitable for examining the heparin sulfate biochemical changes in liver diseases.
【Key words】 heparan sulfate; disaccharide composition; chain length; glycansaminoglycans;
- 【文献出处】 中国现代医学杂志 ,China Journal of Modern Medicine , 编辑部邮箱 ,2007年18期
- 【分类号】Q53
- 【被引频次】1
- 【下载频次】138