Transcriptional suppression of the adrenal cortical peripheral-type benzodiazepine receptor gene and inhibition of steroid synthesis by ginkgolide B.
Treatment of rats and adrenocortical cells with ginkgolide B (GKB), a purified component of Ginkgo biloba leaf extracts, reduces the mRNA, protein, and ligand-binding levels of the adrenal peripheral-type benzodiazepine receptor (PBR), a mitochondrial cholesterol-binding protein, leading to decreased corticosteroid synthesis. In the Y1 adrenocortical cell line, GKB reduced both PBR levels and cyclic AMP-induced steroid formation. In these cells, GKB, but not various steroids and vitamins, reduced the expression of a reporter gene driven by the DNA sequence -624/-513 relative to the transcription start site of the PBR encoding gene. GKB treatment did not affect the SV40 promoter and increased the cytochrome P450 17alpha-hydroxylase gene promoter driven expression of the reporter gene. Electrophoretic mobility shift assays (EMSAs) indicated the presence of a functional transcriptional element bound to the -624/-513 DNA fragment. This GKB-induced inhibition of PBR was mediated by an interaction with a transcription factor that binds to the -636/-616 PBR-promoter region. Deletion or mutation of this sequence eliminated the DNA-protein interaction and the inhibitory effect of GKB on PBR gene transcription. This DNA-binding protein could be detected in nuclear extracts of rat brain, liver, and testis, but not kidney. It is also present in the human adrenal glands. However, the inhibitory effect following GKB treatment could be seen only in the adrenal glands. These results demonstrate that the GKB-activated inhibition of glucocorticoid production is due to a specific transcriptional suppression of the adrenal PBR gene and suggest that GKB might serve as a pharmacological tool to control excess glucocorticoid formation.
Amri
Hakima
H
Division of Hormone Research, Department of Cell Biology, Georgetown University Medical Center, 3900 Reservoir Road, Washington, DC 20057, USA.
Drieu
Katy
K
Papadopoulos
Vassilios
V
D000302:Adrenal Cortex / Q000187:drug effects* / Q000502:physiology; D000818:Animals; D002648:Child; D004224:Diterpenes; D005343:Fibrinolytic Agents / Q000494:pharmacology; D046934:Ginkgolides; D006801:Humans; D007783:Lactones / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008875:Middle Aged; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000235:genetics*; D013256:Steroids / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D014158:Transcription, Genetic / Q000187:drug effects*
D004224:Diterpenes; D005343:Fibrinolytic Agents; D046934:Ginkgolides; D007783:Lactones; D011963:Receptors, GABA-A; D013256:Steroids; C045856:ginkgolide B