Cortisol is a glucocorticoid produced by the zona fasciculata (ZF) of the adrenal cortex. Traditionally, cortisol production and release was seen as being regulated strictly by adrenocorticotropic hormone (ACTH). While this is true of baseline cortisol levels and in response to acute mental stress, the picture is somewhat more complicated in other situations.Interleukin-6 (IL-6) contributes to the maintenance of cortisol levels in situations of prolonged immune or inflammatory stress. AMP activated protein kinase (AMPK) was investigated as a possible mediator of the action of IL-6 or as an independent actor in raising cortisol levels in response to hypoxemic or hypoglycemic stress.5-aminoimidazole-4-carboxamide 1-b-D-ribofuranoside (AICAR) was used to activate AMPK. Bovine ZF tissue fragments were exposed to AICAR alone and together with a known AMPK inhibitor, compound C. Protein or mRNA was then extracted from these tissue fragments. As an indicator of overall steroidogenic activity, these extracts were tested using RT-PCR and western blot assays for relative protein and mRNA levels of steroidogenic acute regulatory (StAR) protein, steroidogenic factor-1 (SF-1), and dosage sensitive sex reversal adrenal hypoplasia congenita gene on the X chromosome, gene 1 (DAX-1). Also a reporter gene assay was performed on H295R cells with a transfected StAR promoter.In bovine ZF tissue fragments, AICAR caused a significant increase of StAR protein and mRNA and SF-1 protein with a decrease of DAX-1 protein in a dose and time dependant manner. DAX-1 mRNA was shown to decrease in response to AICAR administration in a dose dependant manner. AICAR induced increases in StAR protein and SF-1 protein, and the attendant decrease in DAX-1 protein were all shown to be reduced by administration of compound C. This demonstrated that in this situation AICAR is acting through AMPK. When IL-6 was given with compound C the levels of StAR, SF-1, and DAX-1 were significantly reduced from samples treated with IL-6 alone. AICAR exposure also increased StAR promoter activity in a dose and time dependant manner. This AMPK induced increase in steroidogenic activity provides a possible mechanism for increased cortisol during hypoxia and hypoglycemia, and a possible mediator for IL-6 in the ZF.



College and Department

Life Sciences; Physiology and Developmental Biology



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ACTH, Adrenal, Cortex, AMPK, Zona Fasciculata, ZF, Cortisol, StAR, Hypoxia, Hypoglycemia, SF-1, DAX-1, IL-6, Steroidogenesis