SAT269 Orexin Activates Adrenocortical Steroidogenesis With Upregulation Of BMP Signaling By Human H295R Cells

Abstract

Abstract Disclosure: Y. Soejima: None. N. Iwata: None. R. Nishioka: None. M. Honda: None. Y. Nakano: None. K. Yamamoto: None. A. Suyama: None. F. Otsuka: None. Orexins are neuropeptides that express primarily in the hypothalamus and are composed of two isoforms, orexin A and orexin B. Orexin A binds to orexin receptor type 1 (OX1R) and type 2 (OX2R), while orexin B binds selectively to OX2R. Orexin has been reported to have important roles in various systems including sleep-wake regulation and feeding behavior in the central nervous system, whereas its receptors are also expressed in peripheral tissues including the endocrine system. The expression of orexin receptors and the effects of orexins on adrenocortical steroidogenesis have been shown in experiments using adrenocortical cells. Bone morphogenetic proteins (BMPs), which are members of the transforming growth factor (TGF)-β superfamily, also play functional roles in various endocrine tissues. In our previous experiments, we have revealed that BMP-6 enhances aldosterone production which is induced by angiotensin II through activation of mitogen-activated protein kinase (MAPK) signaling. We have also shown that peripheral clock genes have functional roles in modulating adrenocortical steroidogenesis by interacting with adrenocortical BMP system. However, the molecular mechanism of physiological effects of orexin and its interaction with BMP signaling remains unclear. In the present study, we investigated the effects of orexin on adrenal steroidogenesis using human adrenocortical H295R cells, by focusing on the interaction with adrenocortical BMP signaling. First, it was confirmed that both OX1R and OX2R were expressed in H295R cells. It was revealed that orexin A (10-300 nM) stimulation for 24 h directly increased mRNA levels of steroidogenic acute regulatory protein (StAR), cytochrome P450 family 11 subfamily B member 2 (CYP11B2) and cytochrome P450 family 17 subfamily A member 1 (CYP17A1) in a concentration-dependent manner. Orexin A further upregulated the expression of these steroidogenic enzymes in H295R cells co-treated with forskolin (FSK). Next, the interrelationship between orexin and BMP signaling was examined. Stimulation with BMP-6 readily activated Smad1/5/9 phosphorylation in H295R cells and the pretreatment with orexin A for 24 h enhanced the Smad1/5/9 phosphorylation induced by BMP-6. On the other hand, BMP-6 significantly suppressed the expression of OX1R but not that of OX2R. In our previous study, we have reported that BMP-6 upregulates the adrenocortical steroidogenesis via the IP3 and MAPK signaling in H295R cells, suggesting that orexin may have a pivotal role as a functional modulator of BMP-6. Collectively, it was revealed that orexin enhances adrenocortical steroidogenesis by regulating BMP signaling with a feedback system in human adrenocortical cells. Here, we will also discuss the relationship with BMP receptors and the effects on endogenous BMP expression. Presentation: Saturday, June 17, 2023