Detecting cathepsin activity in human osteoarthritis via activity-based probes.
Detecting cathepsin activity in human osteoarthritis via activity-based probes.
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DOI：
10.1186/s13075-015-0586-5
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发表时间：
2015-03-20
影响因子：
4.9
通讯作者：
Dvir-Ginzberg M
中科院分区：
文献类型：
--
作者：
Ben-Aderet L;Merquiol E;Fahham D;Kumar A;Reich E;Ben-Nun Y;Kandel L;Haze A;Liebergall M;Kosińska MK;Steinmeyer J;Turk B;Blum G;Dvir-Ginzberg M
据报道，溶酶体组织蛋白酶与骨关节炎（OA）的病理生理有关，因为它们的促炎条件增加。鉴于组织蛋白酶在OA中的因果作用，监测其特定活性可以提供评估OA严重程度的手段。为此，我们在此试图在体外和体内评估一种基于组织蛋白酶活性的探针（ABP） GB123。在肿瘤坏死因子α (TNFα)和/或白细胞介素1β (IL-1β)刺激下，通过免疫印迹分析和GB123标记，监测培养的原代软骨细胞和条件培养基中组织蛋白酶B和S的蛋白水平和活性。同样，在OA的完整软骨（IC）和降解软骨（DC）区域切片中检测组织蛋白酶活性。最后，用GB123分析无疾病体征、早期OA、晚期OA和类风湿性关节炎（RA）患者的供体滑液（SF）和血清，以检测组织蛋白酶B和s的活性水平。条件培养基外植体和DC切片显示组织蛋白酶B和S的酶活性增强。进一步的组织学分析显示，组织蛋白酶活性在DC浅表区高于IC。检测血清和SF显示，组织蛋白酶B随着OA严重程度在血清和SF中显著升高，但组织蛋白酶S的水平与滑膜炎和RA更相关。根据我们的数据，ABPs监测的组织蛋白酶活性与OA严重程度和关节炎症有很好的相关性，这指向了OA的一个新的病因学靶点，在开发OA早期症状的无创检测方法方面具有重要的转化潜力。本文的在线版本（doi:10.1186/s13075-015-0586-5）包含补充材料，可供授权用户使用。
Lysosomal cathepsins have been reported to contribute to Osteoarthritis (OA) pathophysiology due to their increase in pro-inflammatory conditions. Given the causal role of cathepsins in OA, monitoring their specific activity could provide means for assessing OA severity. To this end, we herein sought to assess a cathepsin activity-based probe (ABP), GB123, in vitro and in vivo. Protein levels and activity of cathepsins B and S were monitored by immunoblot analysis and GB123 labeling in cultured primary chondrocytes and conditioned media, following stimuli with tumor necrosis factor alpha (TNFα) and/or Interleukin 1 beta (IL-1β). Similarly, cathepsin activity was examined in sections of intact cartilage (IC) and degraded cartilage (DC) regions of OA. Finally, synovial fluid (SF) and serum from donors with no signs of diseases, early OA, late OA and rheumatoid arthritis (RA) patients were analyzed with GB123 to detect distinct activity levels of cathepsin B and S. Cathepsin activity in cell lysates, conditioned media explants and DC sections showed enhanced enzymatic activity of cathepsins B and S. Further histological analysis revealed that cathepsin activity was found higher in superficial zones of DC than in IC. Examining serum and SF revealed that cathepsin B is significantly elevated with OA severity in serum and SF, yet levels of cathepsin S are more correlated with synovitis and RA. Based on our data, cathepsin activity monitored by ABPs correlated well with OA severity and joint inflammation, directing towards a novel etiological target for OA, which possesses significant translational potential in developing means for non-invasive detection of early signs of OA. The online version of this article (doi:10.1186/s13075-015-0586-5) contains supplementary material, which is available to authorized users.
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影响因子：
3.7
作者：
Blum G;Weimer RM;Edgington LE;Adams W;Bogyo M
通讯作者：
Bogyo M
影响因子：
4.8
作者：
Cirman, T;Oresic, K;Turk, B
通讯作者：
Turk, B
影响因子：
14.8
作者：
Blum, Galia;von Degenfeld, Georges;Bogyo, Matthew
通讯作者：
Bogyo, Matthew
DOI：
10.1073/pnas.0610811104
发表时间：
2007-06-19
影响因子：
11.1
作者：
Clark, Anna K.;Yip, Ping K.;Malcangio, Marzia
通讯作者：
Malcangio, Marzia
影响因子：
4.1
作者：
BAYLISS, MT;YOUSUFALI, S
通讯作者：
YOUSUFALI, S