Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Siena, Italy.
Dipartimento Scienze Mediche, Chirurgiche e Neuroscienze, Università degli Studi di Siena, Siena, Italy.
Osteoarthritis Cartilage. 2018 Aug;26(8):1078-1086. doi: 10.1016/j.joca.2018.05.017. Epub 2018 May 29.
The aim of this work was to assess baseline serum levels of established biomarkers related to inflammation and oxidative stress in samples from alkaptonuric subjects enrolled in SONIA1 (n = 40) and SONIA2 (n = 138) clinical trials (DevelopAKUre project).
Baseline serum levels of Serum Amyloid A (SAA), IL-6, IL-1β, TNFα, CRP, cathepsin D (CATD), IL-1ra, and MMP-3 were determined through commercial ELISA assays. Chitotriosidase activity was assessed through a fluorimetric method. Advanced Oxidation Protein Products (AOPP) were determined by spectrophotometry. Thiols, S-thiolated proteins and Protein Thiolation Index (PTI) were determined by spectrophotometry and HPLC. Patients' quality of life was assessed through validated questionnaires.
We found that SAA serum levels were significantly increased compared to reference threshold in 57.5% and 86% of SONIA1 and SONIA2 samples, respectively. Similarly, chitotriosidase activity was above the reference threshold in half of SONIA2 samples, whereas CRP levels were increased only in a minority of samples. CATD, IL-1β, IL-6, TNFα, MMP-3, AOPP, thiols, S-thiolated protein and PTI showed no statistically significant differences from control population. We provided evidence that alkaptonuric patients presenting with significantly higher SAA, chitotriosidase activity and PTI reported more often a decreased quality of life. This suggests that worsening of symptoms in alkaptonuria (AKU) is paralleled by increased inflammation and oxidative stress, which might play a role in disease progression.
Monitoring of SAA may be suggested in AKU to evaluate inflammation. Though further evidence is needed, SAA, chitotriosidase activity and PTI might be proposed as disease activity markers in AKU.
本研究旨在评估 SONIA1（n=40）和 SONIA2（n=138）临床试验中纳入的尿黑酸症受试者的基线血清中与炎症和氧化应激相关的既定生物标志物水平。
通过商业 ELISA 检测试剂盒测定血清淀粉样蛋白 A（SAA）、白细胞介素 6（IL-6）、白细胞介素 1β（IL-1β）、肿瘤坏死因子-α（TNFα）、C 反应蛋白（CRP）、组织蛋白酶 D（CATD）、白细胞介素 1 受体拮抗剂（IL-1ra）和基质金属蛋白酶 3（MMP-3）的基线血清水平。通过荧光法测定壳聚糖酶活性。通过分光光度法测定高级氧化蛋白产物（AOPP）。通过分光光度法和 HPLC 测定硫醇、S-硫代蛋白和蛋白硫醇化指数（PTI）。通过验证问卷评估患者的生活质量。
我们发现，与参考阈值相比，SONIA1 和 SONIA2 样本中分别有 57.5%和 86%的 SAA 血清水平显著升高。同样，SONIA2 样本中有一半的壳聚糖酶活性超过参考阈值，而 CRP 水平仅在少数样本中升高。CATD、IL-1β、IL-6、TNFα、MMP-3、AOPP、硫醇、S-硫代蛋白和 PTI 与对照组人群无统计学差异。我们提供的证据表明，SAA、壳聚糖酶活性和 PTI 显著升高的尿黑酸症患者报告生活质量下降的频率更高。这表明尿黑酸症（AKU）症状的恶化与炎症和氧化应激的增加平行，这可能在疾病进展中起作用。
监测 SAA 可能有助于评估 AKU 中的炎症。尽管需要更多的证据，但 SAA、壳聚糖酶活性和 PTI 可能被提议作为 AKU 的疾病活动标志物。
