Identification of α1-Antitrypsin as a Potential Candidate for Internal Control for Human Synovial Fluid in Western Blot.
Identification of α1-Antitrypsin as a Potential Candidate for Internal Control for Human Synovial Fluid in Western Blot.
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DOI：
10.4172/2161-1149.s6-006
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Wei L
中科院分区：
文献类型：
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作者：
Wang S;Zhou J;Wei X;Li P;Li K;Wang D;Wei F;Zhang J;Wei L
滑液蛋白质印迹法已广泛应用于骨关节炎（OA）的研究和诊断，但没有理想的负荷控制。虽然β-肌动蛋白被广泛用作蛋白质印迹中的上样对照，但它不适用于滑液，因为滑液中不需要它作为细胞骨架蛋白。在OA研究中，滑液的良好负荷对照应在正常和OA组的滑液中保持不变，因为滑液蛋白含量可随OA发作时滑膜血管通透性的变化而变化。在本研究中，我们探索了使用α 1-抗胰蛋白酶（A1AT）作为OA滑液的western blot的负载对照的可能性。A1AT水平在炎性病症如类风湿性关节炎（RA）中升高。与RA不同，OA是一种非炎症性疾病，不诱导A1AT。在这项研究中，我们确定A1AT作为一个丰富的组成部分，滑液的质谱和证实，A1AT的水平是相对恒定的人OA和正常滑液之间的蛋白质印迹和ELISA。因此，我们建议，A1AT可能是一个很好的加载控制蛋白质印迹在人类OA滑液研究提供的病理条件，如RA或A1AT缺乏相关的肝脏或肺部疾病被排除。
Western blot of synovial fluid has been widely used for osteoarthritis (OA) research and diagnosis, but there is no ideal loading control for this purpose. Although β-actin is extensively used as loading control in western blot, it is not suitable for synovial fluid because it is not required in synovial fluid as a cytoskeletal protein. A good loading control for synovial fluid in OA studies should have unchanged content in synovial fluids from normal and OA groups, because synovial fluid protein content can vary with changes in synovial vascular permeability with OA onset. In this study, we explore the potential of using α1-antitripsin (A1AT) as loading control for OA synovial fluid in western blot. A1AT level is elevated in inflammatory conditions such as rheumatoid arthritis (RA). Unlike RA, OA is a non-inflammation disease, which does not induce A1AT. In this study, we identified A1AT as an abundant component of synovial fluid by Mass Spectrometry and confirmed that the level of A1AT is relative constant between human OA and normal synovial fluid by western blot and ELISA. Hence, we proposed that A1AT may be a good loading control for western blot in human OA synovial fluid studies provided that pathological conditions such as RA or A1AT deficiency associated liver or lung diseases are excluded.