The endoplasmic reticulum stress - Bcl-2-related ovarian killer (BOK) pathway participates in docosahexaenoic acid-induced adipocyte apoptosis in grass carp (Ctenopharyngodon Idella)

Docosahexaenoic acid (DHA) lessens adipose tissue lipid deposition partly by inducing adipocyte Apoptosis in grass carp, but the underlying mechanism remains unclear. Endoplasmic reticulum (ER) stress and unfolded protein response (UPR) is the novel pathway for inducing Apoptosis. This study aimed to explore the potential role of ER stress in DHA-induced Apoptosis in grass carp (Ctenopharyngodon Idella) adipocytes. DHA-induced activated Apoptosis by deforming the nuclear envelope, condensing the chromatin, and increasing the expression of apoptosis-related proteins and genes in vivo and in vitro (P < 0.05). However, the ER stress inhibitor, 4-phenylbutyric acid (4-PBA), effectively suppressed DHA-induced Apoptosis (P < 0.05), indicating that ER stress mediates DHA-induced adipocyte Apoptosis. Furthermore, we observed that 200 μM DHA significantly up-regulates the transcripts of B cell lymphoma-2 (Bcl-2) related ovarian killer (BOK) in vitro (P < 0.05). BOK is a pro-apoptotic protein in the Bcl-2 Family, which governs the mitochondria Apoptosis pathway. Hence, we hypothesized that BOK might be an important link between ER stress and Apoptosis. We cloned and identified two grass carp BOK genes, BOKa and BOKb, which encode Peptides of 213 and 216 Amino acids, respectively. BOKa primarily localizes in ER and mitochondria in the cytoplasm, while BOKb localizes in the nucleus and cytoplasm of grass carp adipocytes. Moreover, 200 μM DHA treatment up-regulated the mRNA expression of BOKa and BOKb, whereas 4-PBA suppressed the DHA-induced expressions. These results raised the possibility that BOK participates in DHA-induced adipocyte Apoptosis through ER stress signaling, in line with its localization in ER and mitochondria. Two UPR branches, the inositol-requiring Enzyme 1 (IRE1α) and activating transcription factor 6 (ATF6) signaling pathways, are possibly important in DHA-induced adipocyte Apoptosis, unlike protein kinase RNA-activated-like ER kinase (PERK). The study also emphasized the roles of BOKa and BOKb in IRE1α- and ATF6-mediated Apoptosis. This work is the first to elucidate the importance of the ER stress-BOK pathway during adipocyte Apoptosis in teleost.

Apoptosis; BOK; DHA; Endoplasmic reticulum stress; Grass carp.