Della Bruna R, Pinet F, Corvol P, Kurtz A
Physiologisches Institut der Universität Regensburg, Germany.
Kidney Int. 1995 May;47(5):1266-73. doi: 10.1038/ki.1995.181.
A quantitative reverse transcriptase-polymerase chain reaction for mouse renin mRNA was utilized to study the influence of classic second messenger molecules on renin mRNA levels in primary cultures of juxtaglomerular (JG) cells isolated from the kidneys of C57/B16 mice. We found that forskolin (3 microM), an activator of adenylate cyclase led to proportional increases of renin secretion and renin mRNA levels. The nitric oxide (NO) donor, sodium nitroprusside (100 microM), stimulated both renin secretion and renin gene expression, the effect on secretion being stronger than that on renin mRNA levels. An increase of the extracellular concentration of calcium from 0.5 to 3 mM led to a transient inhibition of renin secretion, followed by a marked stimulation of secretion and to a continuous suppression of renin mRNA levels. These were also decreased by the calcium ionophore A 23187 (1 microM). The membrane permeable 8-bromo-cyclic GMP (100 microM) inhibited basal renin secretion without an effect on renin mRNA levels. The phorbol ester phorbol-12-myristate-13-acetate (1 to 100 nM), which was used to stimulate protein kinase C activity, had no significant effects on renin secretion and renin mRNA levels, neither alone nor in combination with forskolin. These findings suggest that cAMP, NO and calcium are effective regulators of renin gene expression in renal JG cells, in a way that cAMP and NO are stimulators and calcium acts as an inhibitor. Moreover, in these acute experiments there appears to be no obligatory link between the secretion and the expression of renin, suggesting that both parameters are separately regulated.
采用定量逆转录聚合酶链反应检测小鼠肾素mRNA，以研究经典第二信使分子对从C57/B16小鼠肾脏分离的球旁（JG）细胞原代培养物中肾素mRNA水平的影响。我们发现，腺苷酸环化酶激活剂福斯可林（3 microM）可使肾素分泌和肾素mRNA水平成比例增加。一氧化氮（NO）供体硝普钠（100 microM）可刺激肾素分泌和肾素基因表达，对分泌的影响强于对肾素mRNA水平的影响。细胞外钙浓度从0.5 mM增加到3 mM会导致肾素分泌短暂抑制，随后分泌显著增加，同时肾素mRNA水平持续受到抑制。钙离子载体A 23187（1 microM）也会降低肾素mRNA水平。膜通透性8-溴环鸟苷酸（100 microM）可抑制基础肾素分泌，但对肾素mRNA水平无影响。用于刺激蛋白激酶C活性的佛波酯佛波醇-12-肉豆蔻酸酯-13-乙酸酯（1至100 nM），单独或与福斯可林联合使用时，对肾素分泌和肾素mRNA水平均无显著影响。这些发现表明，环磷酸腺苷（cAMP）、NO和钙是肾JG细胞中肾素基因表达的有效调节因子，其中cAMP和NO起刺激作用，钙起抑制作用。此外，在这些急性实验中，肾素的分泌与表达之间似乎没有必然联系，这表明这两个参数是分别调节的。
