原发性高血压特征性N-糖基化糖链及相关机制研究

China has the highest mortality rate induced by Essential Hypertension (EH) around the world, moreover, the morbidity rate is rising with years. The pathogenesis of EH is not completely clear at present, in particular, there is no report in the Glycobiology field yet. Our preliminary study shows the significant change of N-glycans in EH patients’ serum. Therefore, patients’ serum and spontaneously hypertensive rats (SHR) as samples, using the high-efficiency and high throughput DSA-FACE technique, the N-glycans of the serum glycoprotein from 400 patients and 50 SHR, and the membrane proteins of impaired heart, brain and kidney from 50 SHR are detected. The characteristic N-glycans of serum glycoprotein and cell surface of impaired target organs during the process of EH are obtained with comparison to healthy control. Using DSA-FACE with exoglycosidases and LC/MS, the structure of characteristic N-glycans is analyzed. The expression of glycosyltransferase genes which are involved in N-glycan synthesis is further studied and the molecular mechanism of abnormal N-glycan modification during the course of EH is investigated. This study increases the knowledge on the pathogenetic mechanism of EH from the point view of Glycobiology. In the meantime, the characteristic N-glycans provide novel potential molecular targets for the treatment of EH and the development of new medicine.

我国是原发性高血压（EH）促发心脑血管疾病死亡率最高的国家，且EH发病率正逐年增高。目前对于该疾病的发病机制还未完全清楚，尤其在糖生物学领域未见研究报道。经初步研究发现EH患者血清N-糖链有明显改变。因此，本项目拟以患者血清及自发性高血压大鼠（SHR）为研究样本，利用高效、高通量的DSA-FACE技术，对400例患者、50例SHR血清糖蛋白及50例SHR受损的心、脑、肾等组织细胞膜蛋白N-糖组进行检测，通过与对照比对分析，筛选获得EH发生时血清糖蛋白及受损靶器官组织细胞表面特征性N-糖基化糖链；以DSA-FACE结合外切糖苷酶、液-质联用技术，对特征性糖链进行结构解析；并进一步分析与糖链合成相关的糖基转移酶基因的表达，初步探讨EH发生过程中N-糖基化修饰异常的分子机制。本研究将从糖生物学角度拓展对EH发生机制的认识，同时特征性N-糖链的获得可为EH的治疗及新药开发提供新的潜在的分子靶标。

我国是原发性高血压（EH）促发心脑血管疾病死亡率最高的国家，且EH发病率还在逐年增高。目前对于该疾病的发病机制还未完全清楚，尤其在糖生物学领域鲜有研究报道。因此，本项目以患者血清及自发性高血压大鼠（SHR）为研究样本，利用DSA-FACE技术，对396例人血清样本（208例EH患者血清和188例无高血压对照血清）、90例大鼠血清（40例SHR血清和50例无高血压对照WKY血清）及140例大鼠心、肾组织细胞膜蛋白N-糖组进行检测。结果发现，1）EH患者血清N-糖组中NGA2FB（Peak2）显著性增高（P<0.01），在1级高血压患者（P=0.004）、尤其是高龄（60岁以上）男性患者血清中显著性增多（P=0.002）；在14周龄SHR大鼠血清中该糖链同样具有显著性增高趋势（P<0.05），表明NGA2FB是EH患者血清中特征性N-糖链；2）NA3FB（Peak10）从单纯收缩性高血压到2级高血压均具有明显降低趋势（P<0.05），且在未发生高血压性心脑血管疾病患者血清中具有显著性（P<0.05）；3）此外，NA2F（Peak6）在3级高血压中极显著降低（P=0），与单纯收缩性高血压、1级、2级高血压患者之间均存在显著性差异（P<0.01），而且在发生靶器官损伤的EH患者中同样具有显著性（P=0.019），表明NA2F为恶性高血压、靶器官受损特征性N-糖链；4）除上述血清中N-糖链具有特异性改变外，经分析140例大鼠心、肾组织细胞表面N-糖组，SHR肾脏组织细胞表面NGA2FB（Peak2）同样也具有显著性增高趋势（P<0.05），说明当EH发生时分泌型糖蛋白（血清中）和驻留型糖蛋白（肾细胞表面）NGA2FB结构修饰异常增加，与EH发生密切相关。本研究从糖生物学角度拓展了对EH发生机制的认识，同时特征性N-糖链的获得可为EH的个体化诊疗及新药开发提供新的潜在分子靶标。