Baldassarre Maurizio, Naldi Marina, Domenicali Marco, Volo Sabrina, Pietra Marco, Dondi Francesco, Caraceni Paolo, Peli Angelo
Center for Applied Biomedical Research (CRBA), S. Orsola-Malpighi Hospital, Bologna, Italy; Department of Medical and Surgical Sciences, University of Bologna, Bologna, Italy.
Center for Applied Biomedical Research (CRBA), S. Orsola-Malpighi Hospital, Bologna, Italy; Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
J Pharm Biomed Anal. 2017 Sep 10;144:263-268. doi: 10.1016/j.jpba.2017.04.041. Epub 2017 Apr 25.
Heat stress has a major impact on veal calves welfare and productivity. Prolonged exposure to warm temperature is associated with several alterations of physiologic processes and increased systemic inflammation and oxidative stress. Bovine serum albumin (BSA) is the most abundant plasma protein and, besides the regulation of osmotic pressure, carries several additional functions, including antioxidant, immunomodulatory, binding and transport activities. Such non-oncotic properties are closely related to structural integrity of the circulating molecule and may be compromised in stressful microenvironments as it occurs in heat stressed animals. Thus, in the present study we developed and validated an LC-MS analytical technique for the characterization of circulating BSA microheterogeneity in veal calves exposed to heat stress. The method was specifically tailored to the structural characteristics of the BSA molecule as well as to the complexity of the biological samples, allowing the identification of several BSA isoforms, each characterized by a specific structural defect. The mass spectrometry based approach enabled the identification of BSA isoforms with reversible and irreversible oxidation and/or glycation and the native BSA, the only isoform endowed with structural and functional integrity. We found that, in veal calves, heat stress is associated to a significant reduction of the native BSA and to a significant increment of the reversibly and irreversibly oxidized BSA. Then, by monitoring the BSA microheterogeneity over a period of moderate heat stress, we found that the native BSA as well as the glycated BSA increased significantly during the recovery period. Based on our results the analysis of the BSA microheterogeneity could represent a novel biomarker for the assessment of animal welfare during environmental stressful conditions.
热应激对犊牛的福利和生产力有重大影响。长时间暴露于温暖温度会导致生理过程的多种改变，并增加全身炎症和氧化应激。牛血清白蛋白（BSA）是最丰富的血浆蛋白，除了调节渗透压外，还具有多种其他功能，包括抗氧化、免疫调节、结合和运输活性。这些非渗透特性与循环分子的结构完整性密切相关，在应激微环境中可能会受到损害，热应激动物的情况就是如此。因此，在本研究中，我们开发并验证了一种液相色谱-质谱分析技术，用于表征热应激犊牛循环中BSA的微观异质性。该方法专门针对BSA分子的结构特征以及生物样品的复杂性进行了调整，能够识别几种BSA异构体，每种异构体都具有特定的结构缺陷。基于质谱的方法能够识别具有可逆和不可逆氧化和/或糖基化的BSA异构体以及天然BSA，天然BSA是唯一具有结构和功能完整性的异构体。我们发现，在犊牛中，热应激与天然BSA的显著减少以及可逆和不可逆氧化的BSA的显著增加有关。然后，通过在中度热应激期间监测BSA的微观异质性，我们发现天然BSA以及糖基化BSA在恢复期显著增加。根据我们的结果，对BSA微观异质性的分析可能代表一种评估环境应激条件下动物福利的新型生物标志物。
