Kankofer M, Wawrzykowski J, Hoedemaker M
Departament of Animal Biochemistry and Physiology, Faculty of Veterinary Medicine, University of Life Sciences, Lublin, Poland.
Reprod Domest Anim. 2013 Aug;48(4):699-704. doi: 10.1111/rda.12152. Epub 2013 Feb 11.
The maintenance of antioxidative/oxidative balance is crucial for cellular and extracellular environment. That is why antioxidative enzymes express their activity in different isoforms in different cell compartments and extracellular space. The aim of study was to verify the results of previous experiment on activities of antioxidative enzymes by the determination of their enzymatic proteins in bovine placental tissues by Western blotting technique. Moreover, the presence of particular isoenzymes was detected and differentiated. Homogenates of maternal and foetal part of both properly released and retained bovine placenta were subjected to PAGE electrophoresis in non-reducing and reducing conditions and Western blotting with appropriate antibodies against superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Electrophoresis allowed for the detection of protein bands of molecular weight related to CuZn-SOD as well as cGSH-Px isoenzymes. The reaction with appropriate antibodies confirmed this. Densitometric analysis, although semi-quantitative, allowed for the observation of trends in differences in antioxidative enzyme proteins, which may partly confirm previously described results in cases of retained and released placenta. Local antioxidative enzymatic mechanisms in bovine placental tissues are represented by CuZn-SOD and cGSH-Px, which show the changes in their expression during improper placental release.
维持抗氧化/氧化平衡对于细胞和细胞外环境至关重要。这就是抗氧化酶在不同细胞区室和细胞外空间以不同同工型表达其活性的原因。本研究的目的是通过蛋白质印迹技术测定牛胎盘组织中抗氧化酶的酶蛋白，以验证先前关于抗氧化酶活性实验的结果。此外，还检测并区分了特定同工酶的存在。对正常排出和滞留的牛胎盘母体和胎儿部分的匀浆在非还原和还原条件下进行聚丙烯酰胺凝胶电泳（PAGE），并用针对超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）的适当抗体进行蛋白质印迹分析。电泳可检测到与铜锌超氧化物歧化酶（CuZn-SOD）以及胞浆谷胱甘肽过氧化物酶（cGSH-Px）同工酶相关的分子量蛋白条带。与适当抗体的反应证实了这一点。密度分析虽然是半定量的，但可以观察到抗氧化酶蛋白差异的趋势，这在一定程度上可能证实了先前关于滞留和排出胎盘情况的描述结果。牛胎盘组织中的局部抗氧化酶机制由CuZn-SOD和cGSH-Px代表，它们在胎盘排出异常时显示出表达变化。
