Argade Sulabha P, Vanichsarn Christopher, Chenoweth Marianne, Parsons C Lowell
Division of Urology, University of California, San Diego Medical Center, San Diego, CA 92103-8897, USA.
BJU Int. 2009 Apr;103(8):1085-9. doi: 10.1111/j.1464-410X.2008.08163.x. Epub 2008 Oct 31.
To confirm abnormal glycosylation of Tamm-Horsfall protein (THP) in patients with interstitial cystitis (IC).
PATIENTS, SUBJECTS AND METHODS: The sialic acid content of THP, a critical component of its biological activity, is reduced in patients with IC. N-glycan shows reduced levels of high molecular weight tri- and tetra-antennary sialylated oligosaccharides. These results are supported by quantitative monosaccharide analysis of neutral and amino sugars in patients vs control subjects. THP was isolated from urine samples of 23 patients with IC and 24 control subjects by salt precipitation. The sialic acid contents were measured using 1,2-diamino-4,5-methylene dioxybenzene-high performance liquid chromatography analysis. For N-glycan profiling, purified THP was treated with peptide:N-glycosidase F to release N-glycans. The purified N-glycans were labelled with 2-aminobenzamide and were profiled by high-pH anion exchange chromatography (HPAEC) with fluorescence detection. The neutral and amino sugars were determined by HPAEC with pulsed amperometric detection.
The total sialic acid in patients was half of that in controls. There was a pattern of reduced level of high molecular weight sialylated oligosaccharide in 17 of 23 patients vs four of 24 controls. The total neutral and amino sugars showed a approximately 30% reduction in patients. The mean (sem) for the controls was 133.79 (6.51) vs 94.76 (6.67) nmol/200 microg of THP for patients (P < 0.001).
THP in patients with IC has reduced sialylation and overall glycosylation, and by inference, THP has a role in the pathophysiology of IC.
证实间质性膀胱炎（IC）患者中Tamm-Horsfall蛋白（THP）存在糖基化异常。
患者、研究对象与方法：IC患者THP生物活性的关键成分唾液酸含量降低。N-聚糖显示高分子量三触角和四触角唾液酸化寡糖水平降低。患者与对照者中性糖和氨基糖的定量单糖分析结果支持了这些发现。通过盐沉淀从23例IC患者和24例对照者的尿液样本中分离出THP。采用1,2-二氨基-4,5-亚甲基二氧基苯-高效液相色谱分析法测定唾液酸含量。对于N-聚糖分析，将纯化的THP用肽：N-糖苷酶F处理以释放N-聚糖。纯化的N-聚糖用2-氨基苯甲酰胺标记，并通过带荧光检测的高pH阴离子交换色谱法（HPAEC）进行分析。通过带脉冲安培检测的HPAEC测定中性糖和氨基糖。
患者的总唾液酸含量是对照者的一半。23例患者中有17例高分子量唾液酸化寡糖水平降低，而24例对照者中只有4例出现这种情况。患者的总中性糖和氨基糖含量降低了约30%。对照者的平均值（标准误）为133.79（6.51）nmol/200μg THP，而患者为94.76（6.67）nmol/200μg THP（P<0.001）。
IC患者的THP唾液酸化和整体糖基化降低，由此推断，THP在IC的病理生理学中起作用。
