Elevation of intracellular cAMP up-regulated thrombomodulin mRNA in cultured vascular endothelial cells derived from spontaneous type-II diabetes mellitus model rat.
To investigate whether antihemostatic function of vascular endothelial cells (VECs) is changed in type-II diabetic model rats, the mRNA expressions of tissue-type and urokinase-type plasminogen activator (t-PA and u-PA), thrombomodulin (TM), PA inhibitor type-1 (PAI-1), and phosphodiesterases (type 3A, 3B, and 4D PDEs) were quantitated by the method of comparative reverse transcriptase-polymerase chain reaction (RT-PCR). VECs from type-II diabetic model Otsuka Long-Evans Tokushima Fatty (OLETF) rats and from its normal counterpart (LETO) rats were cultured for 24 h with dibutyryl adenosine 3',5'-cyclic monophosphate (db-cAMP) or a type-3 PDE inhibitor, cilostazol. Intracellular cAMP concentration was determined by the chemiluminescent enzyme-linked immunosorbent assay (ELISA) system. In cultured VECs from OLETF rats, the basal mRNA expressions of u-PA and TM were significantly decreased as compared to those in cultured VECs from LETO rats. TM mRNA expression in cultured VECs from OLETF rats was increased 2.1-fold at 24 h after treatment with db-cAMP (3 mmol/L). Basal mRNA expressions of type 3A, 3B, and 4D PDEs were significantly higher in VECs from OLETF rats than those from LETO rats. After treatment with cilostazol (30 micromol/L), intracellular cAMP was significantly increased at 60 min and TM mRNA expression was increased 1.5-fold at 24 h. Therefore, elevation of intracellular cAMP by db-cAMP or cilostazol up-regulated TM mRNA expression in cultured VECs from OLETF rats.
Sunagawa
Masanori
M
1st Department of Physiology, Unit of Physiological Science, School of Medicine, University of the Ryukyus, Okinawa, Japan. sunagam@med.u-ryukyu.ac.jp
Shimada
Seiji
S
Hanashiro
Kazuhiko
K
Nakamura
Mariko
M
Kosugi
Tadayoshi
T
D000818:Animals; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology*; D003924:Diabetes Mellitus, Type 2; D042783:Endothelial Cells; D004730:Endothelium, Vascular; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D020334:Rats, Inbred OLETF; D018180:Thrombomodulin / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects*
D012333:RNA, Messenger; D018180:Thrombomodulin; D000242:Cyclic AMP