Interdisciplinary Laboratory of Clinical Analysis Interlab-UMU, Regional Campus of International Excellence Mare Nostrum, University of Murcia, Espinardo, Murcia 30100, Spain.
Interdisciplinary Laboratory of Clinical Analysis Interlab-UMU, Regional Campus of International Excellence Mare Nostrum, University of Murcia, Espinardo, Murcia 30100, Spain; Department of Animal and Food Science, School of Veterinary Medicine, Universitat Autònoma de Barcelona, 08193, Bellaterra, Spain.
Fish Shellfish Immunol. 2018 Apr;75:41-47. doi: 10.1016/j.fsi.2018.01.038. Epub 2018 Jan 31.
A proteomic and biochemical approach was performed to assess the effects of an induced muscle injury on the haemolymph of bivalve molluscs. For this purpose, Mytilus galloprovincialis were exposed to puncture of adductor muscle for three consecutive days, and their haemolymph proteome was then compared to healthy animals using 2-dimensional electrophoresis (2-DE) to identify proteins that differed significantly in abundance. Those proteins were then subjected to tandem mass spectrometry and 6 proteins, namely myosin, tropomyosin, CuZn superoxide dismutase (SOD), triosephosphate isomerase, EP protein and small heat shock protein were identified. SOD and tropomyosin changes were verified by spectrophotometric measurements and western blotting, respectively. As some of the proteins identified are related to muscular damage and oxidative stress, other biomarkers associated with these processes that can be evaluated by automatic biochemical assays were measured including troponin, creatine kinase (CK), and aspartate aminotransferase (AST) for muscle damage, and SOD, trolox equivalent antioxidant capacity (TEAC) and esterase activity (EA) for oxidative stress. Significantly higher concentrations of troponin, CK, AST, and TEAC were observed in mussels after puncture, being also possible biomarkers of non-specific induced damage.
采用蛋白质组学和生物化学方法来评估诱导的肌肉损伤对双壳贝类软体动物血淋巴的影响。为此，将贻贝（Mytilus galloprovincialis）连续暴露于 3 天的肌肉穿刺损伤中，然后使用二维电泳（2-DE）将其血淋巴蛋白质组与健康动物进行比较，以鉴定丰度显著差异的蛋白质。然后对这些蛋白质进行串联质谱分析，鉴定到 6 种蛋白质，即肌球蛋白、原肌球蛋白、CuZn 超氧化物歧化酶（SOD）、磷酸丙糖异构酶、EP 蛋白和小热休克蛋白。通过分光光度法测量和 Western blot 验证了 SOD 和原肌球蛋白的变化。由于鉴定出的一些蛋白质与肌肉损伤和氧化应激有关，因此还测量了其他与这些过程相关的生物标志物，这些生物标志物可以通过自动生化分析进行评估，包括肌钙蛋白、肌酸激酶（CK）和天冬氨酸氨基转移酶（AST）用于肌肉损伤，以及 SOD、trolox 等效抗氧化能力（TEAC）和酯酶活性（EA）用于氧化应激。在穿刺后，贻贝中观察到肌钙蛋白、CK、AST 和 TEAC 的浓度显著升高，这些也是非特异性诱导损伤的生物标志物。
