Dihydroartemisinin exposure impairs porcine ovarian granulosa cells by activating PERK-eIF2α-ATF4 through endoplasmic reticulum stress

Abstract

Dihydroartemisinin (DHA) is an artemisinin derivative commonly used in malaria therapy, and a growing number of studies have focused on the potent anticancer activity of DHA. However, the reproductive toxicity of anticancer drugs is a major concern for young female cancer patients. Previous studies have suggested that DHA can cause embryonic damage and affect oocyte maturation. Here, we explored the side effects of DHA exposure on ovarian somatic cells. We exposed porcine granulosa cells to 5 μM and 40 μM DHA for 24 h or 48 h in vitro. DHA inhibited granulosa cell viability in a dose-dependent manner and, in the 48 h treatment group, DHA enhanced the apoptotic rate. We observed that the levels of intracellular calcium, mitochondrial calcium, and ATP concentration were elevated with DHA treatment. In granulosa cells exposed to DHA, the mRNA levels of endoplasmic reticulum stress-related genes GRP78 and ATF4 were increased. Furthermore, analysis of the unfolded protein response signaling pathway showed that the protein levels of P-PERK, P-eIF2α, and ATF4 were upregulated by DHA exposure. These results demonstrate that in granulosa cells, DHA exposure induces endoplasmic reticulum stress that then activates the PERK/eIF2α/ATF4 signaling pathway, thus providing insight into the mechanism underlying DHA-induced reproductive toxicity, and giving reference to DHA use in females.