title: Osthole, a coumadin analog from Cnidium monnieri (L.) Cusson, stimulates corticosterone secretion by increasing steroidogenic enzyme expression in mouse Y1 adrenocortical tumor cells.
abstract: BACKGROUND
  Osthole is an O-methylated coumadin, which was isolated and purified from the seeds of Cnidium monnieri (L.) Cusson. Osthole is a commonly used traditional Chinese medicine to treat patients with Kidney-Yang deficiency patients, who exhibit clinical signs similar to those of glucocorticoid withdrawal. However, the mechanism of action of osthole is not fully understood.
  
  
  OBJECTIVE
  This study was designed to reveal the effects of osthole on corticosterone production in mouse Y1 cell.
  
  
  METHODS
  Mouse Y1 adrenocortical cells were used to evaluate corticosterone production, which was quantified by enzyme-linked immunosorbent assay (ELISA) kits. Cell viability was tested using the MTT assay, and the mRNA and protein expression of genes encoding steroidogenic enzymes and transcription factors was monitored by quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) and western blotting, respectively.
  
  
  RESULTS
  Osthole stimulated corticosterone secretion from mouse Y1 cells in a dose- and time-dependent manner, and osthole enhanced the effect of dibutyryl-cAMP (Bu2cAMP) on corticosterone production. Further, osthole also increased StAR and CYP11B1 mRNA expression in a dose-dependent manner and enhanced the expression of transcription factors such as HSD3B1, FDX1, POR and RXRα as well as immediate early genes such as NR4A1. Moreover, osthole significantly increased SCARB1(SRB1) mRNA and StAR protein expression in the presence or absence of Bu2cAMP; these proteins are an important for the transport of the corticosteroid precursor cholesterol transport into mitochondria.
  
  
  CONCLUSIONS
  Our results show that the promotion of corticosterone biosynthesis and secretion is a novel effect of osthole, suggesting that this agent can be utilized for the prevention and treatment of Kidney-Yang deficiency syndrome.
authors.affiliation: Basic Medical School of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. Electronic address: pzq527@163.com.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Pan
authors.affiliation: Basic Medical School of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
authors.forename: Zhaoqin
authors.initials: Z
authors.lastname: Fang
authors.affiliation: Basic Medical School of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
authors.forename: Wenli
authors.initials: W
authors.lastname: Lu
authors.affiliation: Basic Medical School of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Liu
authors.affiliation: Basic Medical School of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D015096:3-Hydroxysteroid Dehydrogenases / Q000235:genetics; D000306:Adrenal Cortex Neoplasms / Q000235:genetics / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D031048:Cnidium; D003345:Corticosterone / Q000378:metabolism*; D003374:Coumarins / Q000494:pharmacology*; D051379:Mice; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics; D051122:Scavenger Receptors, Class B / Q000235:genetics; D013252:Steroid 11-beta-Hydroxylase / Q000235:genetics; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: D003374:Coumarins; D010750:Phosphoproteins; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; C454739:Scarb1 protein, mouse; D051122:Scavenger Receptors, Class B; D000098682:Steroidogenic Acute Regulatory Protein; D015096:3-Hydroxysteroid Dehydrogenases; D013252:Steroid 11-beta-Hydroxylase; D003345:Corticosterone; C046627:osthol
keywords: CYP11B1; Corticosterone; Osthole; StAR; Y1 adrenocortical cell