MCU在低氧诱导肉鸡心肌细胞线粒体损伤中的作用

doi:10.11843/j.issn.0366-6964.2021.012.030

Abstract

Abstract: The aim of this study was to investigate whether mitochondrial calcium uniporter(MCU) mediated mitochondrial Ca²⁺ transport was involved in hypoxia-induced mitochondrial damage in broiler cardiomyocytes. In this experiment, the primary cardiomyocytes of chicken embryos of white feathered broilers were isolated and cultured for 24, 48 and 72 h under hypoxic conditions (3%O₂, 5%CO₂, 92%N₂). In order to further determine the role of MCU in mitochondrial injury induced by hypoxia, RU360 was pretreated in the hypoxia group for 72 h to inhibit the expression of MCU. The intracellular Ca²⁺ concentration, mitochondrial Ca²⁺ concentration, mitochondrial reactive oxygen species level and mitochondrial membrane potential were detected by flow cytometry. Gene and protein expressions of MCU and its regulatory factors were detected. The results showed that the purity of cardiomyocytes cultured by differential velocity adherent method could reach over 90%. After 24 h of hypoxic culture, the expression of MCU and MICU1 genes were increased (P<0.05), the concentrations of Ca²⁺ in cytoplasm and mitochondria were significantly increased (P<0.05), and the mitochondrial membrane potential was increased (P<0.05). The expression levels of MCUR1 and MICU1 mRNA were decreased (P<0.05) and intracellular Ca²⁺ concentration was increased (P<0.05) after 48 h hypoxia culture. After 72 h of hypoxic culture, MCU mRNA expression, intracellular and mitochondrial Ca²⁺ were increased (P<0.01), and reactive oxygen species were increased (P<0.01), while mitochondial membrane potential decreased (P<0.01). Compared with the 72 h hypoxic group, the Ca²⁺ in cells and mitochondria decreased, the mitochondrial membrane potential increased and the reactive oxygen species decreased in the RU360 preconditioning group (P<0.01), the expression of MCU mRNA was decreased (P<0.01). The results showed that hypoxia-induced MCU upregulation leads to mitochondrial calcium overload, causing mitochondrial dysfunction and cardiomyocyte injury. Inhibition of MCU expression can reduce hypoxia-induced mitochondrial calcium overload and protect mitochondrial function.

Key words: calcium steady state, hypoxic, MCU, myocardial cells

CLC Number:

S856.2
Q256

PANG Congying, QIAO Na, CHEN Hanming, MA Xinyan, ZHANG Hui, PAN Jiaqiang, TANG Zhaoxin, LI Ying. Role of MCU in Mitochondrial Damage Induced by Hypoxia in Broiler Cardiomyocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3641-3650.

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Role of MCU in Mitochondrial Damage Induced by Hypoxia in Broiler Cardiomyocytes

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