Ganz M B, Pekar S K, Perfetto M C, Sterzel R B
Department of Medicine, West Haven Veterans Administration Medical Center, Yale University School of Medicine, New Haven, Connecticut 06516.
Am J Physiol. 1988 Nov;255(5 Pt 2):F898-906. doi: 10.1152/ajprenal.1988.255.5.F898.
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)
精氨酸加压素（AVP）特异性结合血管平滑肌样系膜细胞（MCs）并影响其收缩。我们测试了这种肽是否也调节大鼠MCs早期传代培养（第2 - 5代）时的生长行为。将亚汇合、血清饥饿的MCs在有或无胰岛素（5微克/毫升）的情况下暴露于AVP（10^(-10) - 10^(-6) M）。为评估DNA复制，在第1、2和3天测定MCs对[3H]胸腺嘧啶核苷的摄取（24小时脉冲）。单独的AVP在24小时时DNA合成平均增加1.97倍，而AVP在48小时和72小时的平均刺激作用分别为7.21倍和5.42倍。同时暴露于AVP和胰岛素的MCs显示出对单独AVP促有丝分裂反应的增强作用。V1受体拮抗剂[1 - （β - 巯基 - β，β - 环亚戊基丙酸），2 - （O - 甲基 - 酪氨酸） - 精氨酸]加压素（PMP）仅抑制AVP诱导的MC生长促进作用（最大抑制率为 - 78.3%）。佛波酯12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯（TPA）急性刺激MC增殖，但不增强AVP的作用。将MCs与600 nM的TPA预孵育48小时可显著抑制AVP诱导的有丝分裂（ - 87.2%）。通过使用fura - 2，采用荧光分光光度法评估细胞内钙（Cai）。加入AVP（10^(-12) - 10^(-6) M）分别导致Cai迅速、短暂、剂量依赖性增加154% - 383%。AVP诱导的Cai增加被3,4,5 - 三甲氧基苯甲酸8 - （二乙氨基）辛酯盐酸盐（TMB - 8）（10^(-8) - 10^(-6) M），一种Cai释放抑制剂（ - 23.9%至 - 72.1%）大大抑制，并且被心房利钠肽AP - 28减弱（ - 38.3%）。（摘要截短于250字）
