肝素与高迁移率族蛋白1相互作用的光谱研究  

Spectroscopic Study on the Interaction of Heparin with High Mobility Group Protein 1

作　　者：凌燕[1] 王春友[1] 杨智勇[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院胰腺外科,湖北武汉430022

出　　处：《分析测试学报》2009年第2期146-149,156,共5页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(30600593)

摘　　要：研究了肝素对高迁移率族蛋白1(HMGB1)构象的影响,探讨了肝素抑制HMGB1促炎活性的机制。采用荧光光谱仪和圆二色谱仪研究终浓度分别为10、50、100、1 000和10 000 u/L的肝素与HMGB1结合的特征,不加肝素做空白对照。荧光光谱显示,加入肝素后,HMGB1的荧光峰值呈先升高再下降的特征,并且随着温度增高荧光强度有所下降;同步荧光光谱显示发射波长发生了红移;圆二色谱显示,HMGB1在加入肝素以后,各种二级结构的组分发生了明显的变化,β折叠的含量相应减少,α螺旋的含量相应增加。当HMGB1为0.025 g/L时,加入50 u/L的肝素对荧光峰值、同步荧光发射波长和二级结构组分的影响最为明显。实验表明肝素与HMGB1作用改变了HMGB1的构象,该作用不与肝素浓度呈正相关。提示肝素通过改变HMGB1构象起到抑制HMGB1促炎活性的作用,该效应可能不与肝素使用剂量呈正相关,使用较低剂量的肝素可能既能获得较强的抗炎效果,又能大幅降低使用肝素导致出血的风险。The effect of heparin on the conformation of high mobility group protein 1 （ HMGBI ） and the mechanism of heparin restraining the pro-inflammatory activity of HMGB1 were investigated. The characteristics of the reaction of HMGB1 with various concentrations of heparin（10, 50, 100, 1 000 and 10 000 u/L） were studied by fluorescence and circular dichroism（CD） spectroscopy. Experimental results showed that the fluorescence intensities of HMGB1 increased with the increase of heparin concentration; then decreased as the heparin concentration increased up to 100 u/L. In addition, the fluorescence intensities decreased with the increase of temperature. Synchronous fluorescence and circular dichroism spectra also showed that the emission wavelength of HMGB1 was red-shifted and the secondary structure of HMGB1 was significantly changed in the presence of heparin： β-pleated sheet structure reduced, whereas α-helix structure increased. Evidently, the reaction of HMGB1 with heparin has changed the conformation of HMGB1, indicating that the pro-inflammatory activity of HMGB1 could be restrained through varying the conformation of HMGB1. However, the restraint was not positively con＇elated with the concentration of heparin. By using low dose of heparin, the anti-inflammatory effect would be more effective and the risk of bleeding was substantially reduced.

关 键 词：高迁移率族蛋白1 肝素 荧光光谱 荧光增强 圆二色光谱 

分 类 号：O657.3[理学—分析化学] R973.2[理学—化学]