Imaging NAD(H) Redox Alterations in Cryopreserved Alveolar Macrophages from Ozone-Exposed Mice and the Impact of Nutrient Starvation during Long Lag Times

Antioxidants

HeN. Xu¹  LinZ. Li¹  Joanna Floros²  Shaili Amatya³ 

[1] Britton Chance Laboratory of Redox Imaging, Department of Radiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA;Departments of Pediatrics and Obstetrics and Gynecology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA;Division of Neonatal-Perinatal Medicine, Center for Host Defense, Inflammation, Lung Diseases (CHILD), Department of Pediatrics, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA;

关键词: bronchoalveolar lavage (BAL);    fresh alveolar macrophages;    surfactant protein A (SP-A);    innate immune responses;    oxidative stress;    ROS;   

DOI  :  10.3390/antiox10050767

来源: DOAJ

【 摘 要 】

Employing the optical redox imaging technique, we previously identified a significant redox shift of nicotinamide adenine dinucleotide (NAD and the reduced form NADH) in freshly isolated alveolar macrophages (AM) from ozone-exposed mice. The goal here was twofold: (a) to determine the NAD(H) redox shift in cryopreserved AM isolated from ozone-exposed mice and (b) to investigate whether there is a difference in the redox status between cryopreserved and freshly isolated AM. We found: (i) AM from ozone-exposed mice were in a more oxidized redox state compared to that from filtered air (FA)-exposed mice, consistent with the results obtained from freshly isolated mouse AM; (ii) under FA exposure, there was no significant NAD(H) redox difference between fresh AM that had been placed on ice for 2.5 h and cryopreserved AM; however, under ozone exposure, fresh AM were more oxidized than cryopreserved AM; (iii) via the use of nutrient starvation and replenishment and H₂O₂-induced oxidative stress of an AM cell line, we showed that this redox difference between cryopreserved and freshly isolated AM is likely the result of the double “hit”, i.e., the ozone-induced oxidative stress plus nutrient starvation that prevented freshly isolated AM from a full recovery after being on ice for a prolonged time period. The cryopreservation technique we developed eliminates/minimizes the effects of oxidative stress and nutrient starvation on cells. This method can be adopted to preserve lung macrophages from animal models or clinical patients for further investigations.

【 授权许可】

Unknown