Virgilio Tommaso, Latino Irene, Pizzichetti Chiara, Chahine Kamil, Capucetti Arianna, Detotto Carlotta, Bergadano Alessandra, F Gonzalez Santiago
Institute for Research in Biomedicine, Università della Svizzera italiana, Bellinzona, Switzerland.
Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland.
Lab Anim (NY). 2025 Sep 18. doi: 10.1038/s41684-025-01614-4.
Anesthesia is indispensable for minimizing stress during invasive procedures. However, anesthesia can induce undesired effects on the organism and its physiological homeostasis. In experiments involving animals, these consequences may reduce animal welfare and alter experimental outcomes. Moreover, most of the studies characterizing the effects of murine anesthetic protocols on animal welfare do not explore the impact of prolonged anesthesia for procedures lasting more than 2 h. Here we investigated the effect of prolonged anesthesia on vital parameters and immune responses to vaccination, comparing isoflurane (Iso), ketamine-xylazine (KX), and KX with oxygen supplementation (KXO), in the presence or absence of buprenorphine (BPP). KX induced hypoxia and 100% mortality, which were prevented by oxygen supplementation (KXO) and were not associated with BPP. By contrast, Iso induced safe and fast induction and recovery. Furthermore, we investigated the effects of these protocols on the immune responses and motility of immune cells following vaccination. The results showed that KX reduced immune cell numbers and increased cell death, complemented by elevated levels of the inflammatory proteins IL-6 and IFNγ. In addition, KX altered the motility patterns of T cells, B cells and neutrophils, potentially influenced by hypoxia. Conversely, Iso exhibited fewer immune artifacts, regardless of BPP. These findings highlight the importance of evaluating anesthesia protocols with respect to animal welfare and experimental reproducibility, especially for immunological studies. Oxygen supplementation emerged as an important refinement to mitigate hypoxia in KX. Iso showed superior safety and fewer artifacts compared with KX and KXO, demonstrating its suitability for immunological research and intravital microscopy.
麻醉对于在侵入性操作过程中最大程度减少应激至关重要。然而，麻醉会对机体及其生理稳态产生不良影响。在涉及动物的实验中，这些后果可能会降低动物福利并改变实验结果。此外，大多数描述小鼠麻醉方案对动物福利影响的研究并未探讨持续超过2小时的长时间麻醉对实验的影响。在此，我们研究了长时间麻醉对重要参数以及疫苗接种免疫反应的影响，比较了异氟烷（Iso）、氯胺酮-赛拉嗪（KX）以及补充氧气的氯胺酮-赛拉嗪（KXO）在有无丁丙诺啡（BPP）情况下的作用。KX会导致缺氧和100%的死亡率，补充氧气（KXO）可预防这些情况，且与BPP无关。相比之下，Iso诱导安全且快速的麻醉诱导和苏醒。此外，我们研究了这些方案对疫苗接种后免疫反应和免疫细胞运动性的影响。结果显示，KX减少了免疫细胞数量并增加了细胞死亡，同时炎症蛋白IL-6和IFNγ水平升高。此外，KX改变了T细胞、B细胞和中性粒细胞的运动模式，这可能受缺氧影响。相反，无论有无BPP，Iso表现出的免疫干扰较少。这些发现凸显了评估麻醉方案对于动物福利和实验可重复性的重要性，尤其是对于免疫学研究。补充氧气成为减轻KX中缺氧的一项重要改进措施。与KX和KXO相比，Iso显示出更高的安全性和更少的干扰，表明其适用于免疫学研究和活体显微镜检查。
