Escribano Damián, Tvarijonaviciute Asta, Tecles Fernando, Cerón José J
Interdisciplinary Laboratory of Clinical Analysis, Interlab-UMU, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Espinardo, Murcia, Spain.
Interdisciplinary Laboratory of Clinical Analysis, Interlab-UMU, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Espinardo, Murcia, Spain.
Vet Immunol Immunopathol. 2015 Feb 15;163(3-4):210-5. doi: 10.1016/j.vetimm.2014.12.002. Epub 2014 Dec 16.
Paraoxonase 1 (PON1) is a serum enzyme synthesised and secreted primarily by the liver. It possesses anti-inflammatory properties limiting the production of pro-inflammatory mediators. The objectives of this study were to validate three spectrophotometric assays for the quantification of PON1 activity in pig serum, and to determine if PON1 activity in porcine behaves as a negative acute phase protein (APP), decreasing in inflammatory conditions. An analytical validation using three different substrates - 5-thiobutil butyrolactone (TBBL), phenylacetate (PA) and 4-(p)-nitrophenyl acetate (pNA) - was performed. In addition, inflammation was experimentally induced in five pigs by subcutaneous injection of turpentine oil, while five control pigs were left untreated. The treated pigs showed significant increases in CRP and decreases in albumin, indicating an inflammatory condition. The three substrates used would be suitable for PON1 activity measurements in serum samples, since they offer adequate precision (coefficients of variation<10%), sensitivity (0.01, 0.15, 0.02 U/mL for TBBL, pNA and PA respectively) and accuracy (r=0.99). In addition, PON1 behaves as a negative APP in pigs since a significant decrease (P<0.05) in its activity after 72 h of the induction of the inflammation was observed with all substrates.
对氧磷酶1（PON1）是一种主要由肝脏合成并分泌的血清酶。它具有抗炎特性，可限制促炎介质的产生。本研究的目的是验证三种分光光度法用于定量猪血清中PON1活性，并确定猪体内的PON1活性是否表现为负急性期蛋白（APP），即在炎症条件下降低。使用三种不同的底物——5-硫代丁基丁内酯（TBBL）、苯乙酸盐（PA）和4-（对）-硝基苯乙酸盐（pNA）进行了分析验证。此外，通过皮下注射松节油在五头猪中实验性诱导炎症，而五头对照猪不进行处理。处理后的猪显示CRP显著升高，白蛋白降低，表明处于炎症状态。所使用的三种底物适用于血清样本中PON1活性的测量，因为它们具有足够的精密度（变异系数<10%）、灵敏度（TBBL、pNA和PA分别为0.01、0.15、0.02 U/mL）和准确度（r = 0.99）。此外，PON1在猪体内表现为负急性期蛋白，因为在炎症诱导72小时后，使用所有底物均观察到其活性显著降低（P<0.05）。
