AIM The study aims to investigate the role of Fas-activated serine/threonine kinase (FASTK) in hypoxia/reoxygenation (H/R)-induced mitochondrial respiratory dysfunction in cardiomyocytes.
METHODS Primary cardiomyocytes were isolated from 1 to 2-day-old C57BL6/J mice and were in vitro cultured. Cardiomyocytes were transfected with adenovirus vectors carrying control green fluorescent protein (Ad-Control) or full-length FASTK (Ad-FASTK). Mitochondrial respiratory function was evaluated by the Seahorse metabolic analyzer. Cardiomyocyte death was evaluated by caspase-3 activity, lactate dehydrogenase (LDH) release, and MTT cell viability assay.
RESULTS H/R obviously suppressed FASTK mRNA and protein expression in cardiomyocytes (all P<0.01). Compared with Ad-Control, Ad-FASTK significantly increased FASTK mRNA and protein expression levels. Overexpression of FASTK significantly preserved mitochondrial respiratory function in H/R-exposed cardiomyocytes, as evidenced by higher basal and maximal respiratory capability as well as stronger adenosine triphosphate (ATP) production (all P<0.01). Moreover, FASTK overexpression ameliorated H/R-induced caspase-3 activation, LDH release, and cardiomyocyte death (all P<0.01).
CONCLUSION Overexpression of FASTK ameliorates H/R-associated mitochondrial respiratory dysfunction and cell death in cardiomyocytes.
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