Role of reactive oxygen species in hypoxia postconditioning-induced activation of Nrf2/ARE signaling pathway in rat cardiomyocytes
10.3760/cma.j.cn131073.20211018.00511
- VernacularTitle:活性氧在缺氧后处理激活大鼠心肌细胞Nrf2/ARE信号通路中的作用
- Author:
Wenjing ZHOU
1
;
Kun TANG
;
Peng XU
;
Tian YU
;
Haiying WANG
Author Information
1. 遵义医科大学附属医院贵州省麻醉与器官保护基础研究重点实验室，遵义 563000
- Keywords:
NF-E2-related factor 2;
Response elements;
Reactive oxygen species;
Reperfusion injury;
Myocytes, cardiac;
Hypoxic postconditioning
- From:
Chinese Journal of Anesthesiology
2022;42(5):560-564
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the role of reactive oxygen species (ROS) in hypoxia postconditioning-induced activation of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway in rat cardiomyocytes.Methods:Primary cardiomyocytes of adult rats were isolated and cultured and divided into 4 groups ( n=20 each) using a random number table method: normal group (group N), hypoxia-reoxygenation group (group HR), hypoxia postconditioning group (group HPO) and hypoxia postconditioning plus an ROS scavenger N-(2-Amidinopropionyl)-glycine (MPG) group (group HPO+ MPG). Cells were exposed to hypoxia for 45 min followed by 60 min reoxygenation to develop the cardiomyocyte hypoxia-reoxygenation injury model.In HPO group, cells were subjected to 3 cycles of 5-min hypoxia/5-min reoxygenation after 45 min hypoxia, followed by reoxygenation for 60 min.In HPO+ MPG group, MPG (final concentration 2 mmol/L) was added at 35 min of hypoxia, cells were subjected to hypoxia for 10 min, and the other treatments were similar to those previously described in group HPO.At the end of reoxygenation, the intracellular calcium level and Nrf2 activity were measured, the ultrastructure of cardiomyocytes was observed, and the Flameng score of mitochondria was assessed, and the expression of Nrf2, NAD(P)H quinine oxidoreductase 1 (NQO1), superoxide dismutase 1 (SOD1) and heme oxygenase-1 (HO-1) protein and mRNA was detected using real-time polymerase chain reaction and Western blot. Results:Compared with group N, the intracellular free Ca 2+ level, Nrf2 activity and Flameng score were significantly increased, and the expression of Nrf2, NQO1, SOD1 and HO-1 protein and mRNA was down-regulated in group HR ( P<0.05). Compared with group HR, the intracellular free Ca 2+ level and Flameng score were significantly decreased, the Nrf2 activity was increased, and the expression of Nrf2, NQO1, SOD1 and HO-1 protein and mRNA was up-regulated in group HPO ( P<0.05). Compared with group HPO, the intracellular free Ca 2+ level and Flameng score were significantly increased, the Nrf2 activity was decreased, and the expression of Nrf2, NQO1, SOD1 and HO-1 protein and mRNA was down-regulated in group HPO+ MPG ( P<0.05). Conclusions:The mechanism by which hypoxia postconditioning activates the Nrf2/ARE signaling pathway in rat cardiomyocytes may be related to ROS.
