Heat stress induces oxidative stress and activates the KEAP1-NFE2L2-ARE pathway in reproduction-related cells

Abstract

Heat stress negatively affects the reproductive function of in animals and humans. Although a relationship between heat and oxidative stress has been suggested, the underlying mechanism has not been sufficiently examined in reproduction-related cells. Therefore, we aimed to investigate whether heat stress induces oxidative stress using a variety of reproduction-related cells including bovine placental and cumulus–granulosa cells, human cell lines derived from cervical and endometrial cancers, and fibroblasts derived from endometrium. Quantitative polymerase chain reaction analysis showed that the expression levels of representative heat and oxidative stress–related genes were significantly increased in cells cultured at high temperatures compared with those in cells cultured at basal temperatures. Moreover, luciferase reporter assays showed that the reporter activity of the heat shock element and antioxidant responsive element (ARE) was increased in cells cultured at high temperatures compared with that in cells cultured at basal temperatures. Furthermore, the stability of nuclear factor erythroid 2 like 2 (NFE2L2), a master regulator of the cellular stress response, increased under high temperatures. Point mutations in Kelch-like ECH-associated protein 1 (KEAP1) cysteine residues reduced the luciferase activity. Our results suggest that heat stress induces oxidative stress and that the KEAP1-NFE2L2-ARE pathway may play a protective role in reproduction-related cells against heat stress.