Mechanism for aldosterone potentiation of angiotensin II-stimulated rat arterial smooth muscle cell proliferation.
After earlier studies in which secretion of aldosterone was demonstrated to be important in rat arterial smooth muscle cell (RASMC) proliferation in vitro, the presence of both 11beta-hydroxylase (CYP11B1) and aldosterone synthase (CYP11B2) gene transcription were shown in these cells by real-time reverse transcription-polymerase chain reaction (RT-PCR). In proliferation studies, tritiated thymidine incorporation into RASMC and RASMC cell number were both significantly increased by angiotensin II (Ang II) (10(-7) mol/L) compared with controls (P<0.01), but this effect was inhibited by the 3beta-hydroxysteroid-dehydrogenase inhibitor trilostane (10(-6) mol/L and 10(-5) mol/L, P<0.05). Aldosterone alone added to RASMC did not significantly change tritiated thymidine incorporation when compared with controls, but the Ang II-induced increase was significantly enhanced by aldosterone at 10(-10) mol/L and 10(-8) mol/L (P<0.05). Neither corticosterone nor 18-hydroxydeoxycorticosterone had any such potentiating effect. RT-PCR analysis and real-time quantitative RT-PCR revealed an increase of Ang II type-1 (AT1) receptor mRNA in RASMC treated by aldosterone (10(-8) mol/L) compared with untreated controls, and this was correlated with a small but significant increase in AT1 receptor protein (P<0.05), as assessed by immunoblotting analysis. These data confirm that steroid production by RASMC is critical in the response to Ang II, and the data support the view that aldosterone specifically is required for the full proliferative response to Ang II in RASMC. One way it may act is by modulating the expression and functions of the AT1 receptor.
School of Biological Sciences, Queen Mary, University of London, Mile End Road, London E1 4NS, UK. f.xiao@qmul.ac.uk
Fang
F
Xiao
John R
JR
Puddefoot
Stewart
S
Barker
Gavin P
GP
Vinson
D000450:Aldosterone / Q000494:pharmacology*; D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D003345:Corticosterone / Q000494:pharmacology; D019405:Cytochrome P-450 CYP11B2 / Q000096:biosynthesis / Q000235:genetics; D004261:DNA Replication / Q000187:drug effects; D003900:Desoxycorticosterone / Q000031:analogs & derivatives* / Q000494:pharmacology; D004357:Drug Synergism; D020869:Gene Expression Profiling; D008297:Male; D008954:Models, Biological; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D044140:Receptor, Angiotensin, Type 1 / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013252:Steroid 11-beta-Hydroxylase / Q000096:biosynthesis / Q000235:genetics; D013268:Stimulation, Chemical; D014158:Transcription, Genetic / Q000187:drug effects*; D015383:Zona Fasciculata / Q000166:cytology / Q000378:metabolism; D015384:Zona Glomerulosa / Q000166:cytology / Q000378:metabolism
D012333:RNA, Messenger; D044140:Receptor, Angiotensin, Type 1; D000804:Angiotensin II; C042509:18-hydroxydeoxycorticosterone; D003900:Desoxycorticosterone; D000450:Aldosterone; D019405:Cytochrome P-450 CYP11B2; D013252:Steroid 11-beta-Hydroxylase; D003345:Corticosterone