Campos-Sánchez Jose Carlos, Esteban María Ángeles, Guardiola Francisco A
Immunobiology for Aquaculture Group, Department of Cell Biology and Histology. Faculty of Biology, Campus Regional de Excelencia Internacional "Campus Mare Nostrum", University of Murcia, 30100, Murcia, Spain.
Immunobiology for Aquaculture Group, Department of Cell Biology and Histology. Faculty of Biology, Campus Regional de Excelencia Internacional "Campus Mare Nostrum", University of Murcia, 30100, Murcia, Spain.
Fish Shellfish Immunol. 2025 Feb;157:110084. doi: 10.1016/j.fsi.2024.110084. Epub 2024 Dec 9.
Proteinogram is a semiquantitative method specially used in clinic to separate the serum proteins from patients for use in the diagnosis of diseases. However, this methodology has only been applied very recently with this approach in farmed fish. Thus, the aim of this study was to explore the changes in the serum proteinogram of gilthead seabream (Sparus aurata), after triggering an acute or chronic sterile inflammation. For this, two experiments were carried out: i) Acute inflammation experiment: seabream specimens were injected intramuscularly with 50 μL of λ-carrageenan (0.5 mg fish) or buffer (control) and blood samples were collected at 3, 6 and 24 h post-injection; ii) Chronic inflammation experiment: specimens were injected at 0, 7 and 14 days with 500, 250 and 250 μL of λ-carrageenan, respectively (20 mg fish) or buffer, and blood samples were collected at 15 days post-injection. In both cases, serum was obtained and processed by electropherograms and HPLC-mass spectrometry. Results of electropherograms of control fish revealed four major proteins of 19.5, 76.3, 104.4, and 156.7 kDa in the serum. These four proteins were correlated with apolipoprotein A-II (II (the counterpart of mammalian albumin, Apo fraction), serotransferrin (β fraction), inter-α-trypsin inhibitor heavy chain H3-like (α1 fraction) and α-2-macroglobulin-like (α2 fraction) according to the results obtained with HPLC-mass spectrometry. In a statistical view (p < 0.05), no variations were detected in the four major serum protein bands between the control and the acutely inflamed groups. However, in chronically inflamed fish, the Apo fraction decreased statistically compared to the control group. In contrast, the α1 and α2 fractions were statistically increased in the serum of fish sampled 15 days after λ-carrageenan injection, compared to those found in the control fish. α1 and α2 protein fractions are recognized indicators of inflammation in mammals. Consequently, our study presents a novel method for assessing both acute and chronic λ-carrageenan-induced sterile inflammation in gilthead seabream, which could be applicable to other marine species for diagnostic purposes.
蛋白质图谱是一种专门用于临床的半定量方法，用于分离患者血清蛋白以辅助疾病诊断。然而，这种方法最近才应用于养殖鱼类。因此，本研究的目的是探究在引发急性或慢性无菌炎症后，金头鲷血清蛋白质图谱的变化。为此，进行了两个实验：i）急性炎症实验：给金头鲷样本肌肉注射50μL λ-角叉菜胶（0.5mg/鱼）或缓冲液（对照组），并在注射后3、6和24小时采集血样；ii）慢性炎症实验：分别在第0、7和14天给样本注射500、250和250μL λ-角叉菜胶（20mg/鱼）或缓冲液，并在注射后15天采集血样。在这两种情况下，均获取血清并通过电泳图谱和高效液相色谱-质谱联用进行处理。对照鱼的电泳图谱结果显示血清中有四种主要蛋白质，分子量分别为19.5、76.3、104.4和156.7kDa。根据高效液相色谱-质谱联用的结果，这四种蛋白质分别与载脂蛋白A-II（II（哺乳动物白蛋白的对应物，载脂蛋白部分）、血清转铁蛋白（β部分）、α-胰蛋白酶抑制剂重链H3样蛋白（α1部分）和α-2-巨球蛋白样蛋白（α2部分）相关。从统计学角度来看（p<0.05），对照组和急性炎症组之间的四条主要血清蛋白带未检测到变化。然而，在慢性炎症的鱼中，与对照组相比，载脂蛋白部分在统计学上有所下降。相反，与对照鱼相比，在注射λ-角叉菜胶15天后采集的鱼血清中，α1和α2部分在统计学上有所增加。α1和α2蛋白部分是哺乳动物炎症的公认指标。因此，我们的研究提出了一种评估λ-角叉菜胶诱导的金头鲷急性和慢性无菌炎症的新方法，该方法可能适用于其他海洋物种的诊断目的。
