Arginine vasopressin promotes growth of rat glomerular mesangial cells in culture.
Arginine vasopressin (AVP) binds specifically to vascular smooth muscle-like mesangial cells (MCs) and affects contraction. We tested whether this peptide also modulates growth behavior of rat MCs in early subculture (passage 2-5). Subconfluent, serum-starved MCs were exposed to AVP (10(-10)-10(-6) M) in the presence or absence of insulin (5 micrograms/ml). To assess DNA replication, MC uptake of [3H]thymidine (24-h pulse) was determined on days 1, 2, and 3. AVP alone averaged a 1.97-fold increase in DNA synthesis at 24 h, whereas the mean stimulatory effects of AVP at 48 and 72 h were 7.21- and 5.42-fold, respectively. MCs exposed simultaneously to AVP and insulin showed potentiation of the mitogenic response to AVP alone. The V1-receptor antagonist [1-(beta-mercapto-beta,beta-cyclopentamethylene proprionic acid), 2-(O-methyl-Tyr)-Arg]vasopressin (PMP) inhibited only AVP-induced promotion of MC growth (maximal inhibition of -78.3%). The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA) acutely stimulated MC proliferation but did not add to the AVP effect. Preincubation of MCs with 600 nM of TPA for 48 h significantly inhibited AVP-induced mitogenesis (-87.2%). By use of fura-2, intracellular calcium (Cai) was assessed by spectrofluorometry. The addition of AVP (10(-12)-10(-6) M) led to a rapid, transient, dose-dependent increase in Cai of 154-383%, respectively. The AVP-induced increase in Cai was greatly inhibited by 3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester hydrochloride (TMB-8) (10(-8)-10(-6) M), an inhibitor of Cai release (-23.9 to -72.1%), and it was blunted by the atrial natriuretic peptide AP-28 (-38.3%).(ABSTRACT TRUNCATED AT 250 WORDS)
Ganz
M B
MB
Department of Medicine, West Haven Veterans Administration Medical Center, Yale University School of Medicine, New Haven, Connecticut 06516.
Pekar
S K
SK
Perfetto
M C
MC
Sterzel
R B
RB
D000818:Animals; D001127:Arginine Vasopressin / Q000031:analogs & derivatives / Q000494:pharmacology*; D009320:Atrial Natriuretic Factor / Q000494:pharmacology; D001572:Benzofurans; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004357:Drug Synergism; D005456:Fluorescent Dyes; D016257:Fura-2; D005707:Gallic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D005920:Glomerular Mesangium / Q000166:cytology*; D007328:Insulin / Q000494:pharmacology; D008297:Male; D051381:Rats; D013050:Spectrometry, Fluorescence; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D001572:Benzofurans; D002121:Calcium Channel Blockers; D005456:Fluorescent Dyes; D007328:Insulin; D001127:Arginine Vasopressin; C006014:8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate; D005707:Gallic Acid; C025043:vasopressin, 1-(1-mercaptocyclohexaneacetic acid)-2-(O- methyl-L-tyrosine)-8-L-arginine-; D009320:Atrial Natriuretic Factor; D004247:DNA; D014700:Verapamil; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium; D016257:Fura-2