Cellular signaling mechanisms for stimulation of growth hormone secretion and growth hormone primary transcripts by immunosuppressant agents, FK506 and cyclosporin A, in cultured rat pituitary cells.
Although an immunosuppressant, FK506, has been known to stimulate growth hormone (GH) release from rat somatotropes, the cellular signaling mechanism is unknown. In the present study, intracellular signaling pathways were investigated for FK506- and cyclosporin A (CsA)-induced GH release in cultured rat anterior pituitary cells. Northern and Western blot analysis revealed that the FK506-binding protein (FKBP12) and the CsA-binding protein (cyclophilin A) exist at the mRNA and protein level in the rat anterior pituitary tissue. FK506 and CsA increased GH release in a dose-dependent manner and inhibited calcineurin (CaN) activity in the cultured pituitary cells. The third immunosuppressant, rapamycin (RP), inhibited the FK506-induced GH release, although RP alone had no effect. Protein kinase A (PKA) inhibitors, H-89 and HA-1004 and EGTA blocked FK506- and CsA-induced GH release. TGF-beta did not alter basal GH release, but inhibited FK506-induced GH release. GH primary transcripts were increased by FK506, and the effects were blocked by H-89 and HA-1004. These results suggest that the immunosuppressants, FK506 and CsA, stimulate GH release by inhibiting CaN activity which results in the activation of the PKA system in the rat somatotropes. TGF-beta receptors might be involved in FK506-induced GH release as a separate pathway. FK506 also stimulates GH primary transcripts via a PKA-dependent mechanism in a manner similar to its effects on GH release.
First Department of Internal Medicine, Kagawa Medical University, Ikenobe, Japan.
H
H
Ohye
M
M
Sato
K
K
Murao
S
S
Matsubara
M
M
Tokuda
J
J
Takahara
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D001933:Brain Stem / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D019703:Calcineurin / Q000378:metabolism; D002531:Cerebellum / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D002540:Cerebral Cortex / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D016572:Cyclosporine / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects / Q000276:immunology; D013006:Growth Hormone / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007031:Hypothalamus / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D020104:Immunophilins / Q000032:analysis / Q000235:genetics; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007546:Isoquinolines / Q000494:pharmacology; D019169:Jurkat Cells / Q000737:chemistry / Q000201:enzymology; D008297:Male; D019696:Peptidylprolyl Isomerase / Q000032:analysis / Q000235:genetics; D010902:Pituitary Gland / Q000737:chemistry / Q000166:cytology / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000276:immunology*; D020123:Sirolimus / Q000494:pharmacology; D013449:Sulfonamides; D016559:Tacrolimus / Q000494:pharmacology*; D022021:Tacrolimus Binding Proteins; D015533:Transcriptional Activation / Q000187:drug effects / Q000276:immunology
D004791:Enzyme Inhibitors; D007166:Immunosuppressive Agents; D007546:Isoquinolines; D012333:RNA, Messenger; D013449:Sulfonamides; D016572:Cyclosporine; D013006:Growth Hormone; C043204:N-(2-guanidinoethyl)-5-isoquinolinesulfonamide; D019703:Calcineurin; D022021:Tacrolimus Binding Proteins; D020104:Immunophilins; D019696:Peptidylprolyl Isomerase; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D020123:Sirolimus; D016559:Tacrolimus