Cole F E, Rovigatti U, Iwata T, Vaughn J, Frohlich E D
Alton Ochsner Medical Institutions, New Orleans, Louisiana.
Hypertension. 1989 Jun;13(6 Pt 2):799-803. doi: 10.1161/01.hyp.13.6.799.
The natriuretic effects of atrial peptide hormones have been attributed, at least in part, to their stimulation of guanylate cyclase activity in renal cell membranes. The effects of atrial natriuretic factor (ANF) on stimulation of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) accumulation were investigated in cloned human kidney tumor (hKT) cells and parent cells from a human renal tumor epithelial cell line (SK-NEP-1). Human ANF-(99-126) (10(-6)M) stimulated (p less than 0.001) cellular cGMP accumulation in a dose-dependent manner from a basal level of 0.26 +/- 0.04 to 3.73 +/- 0.81 pmol/mg protein/5 mi (mean +/- SEM, n = 13). ANF stimulation of cGMP accumulation was specific, in that high concentrations (10(-6)M) of atriopeptin I [rat ANF-(103-123)], angiotensin II, arginine vasopressin, and amiloride (10(-4)M) did not increase basal cGMP. Amiloride (10(-4)M) enhanced (p less than 0.01, n = 6) the ANF stimulation of cGMP accumulation (1.24 +/- 0.39 pmol/mg protein/5 min), particularly at low doses of ANF (10(-10)M) where stimulation by ANF without amiloride (0.34 +/- 0.08 pmol/mg protein/5 min) was barely distinguishable from a basal level (0.19 +/- 0.02 pmol/mg protein/5 min) of cGMP accumulation. The stimulatory effect of ANF (1.59 +/- 0.07 pmol/mg protein/5 min) was attenuated (0.75 +/- 0.06 pmol/mg protein/5 min, p less than 0.01, n = 6) by preincubation of the cells with pertussis toxin but not by cholera toxin. ANF (4.56 +/- 0.93 pmol/mg protein/5 min, n = 8) did not affect cAMP accumulation (4.32 +/- 0.98 pmol/mg protein/5 min) in hKT cells. This is the first report of an ANF responsive human renal cell line, and its use should facilitate investigation of ANF-receptor interactions.
心房肽激素的利钠作用至少部分归因于它们对肾细胞膜中鸟苷酸环化酶活性的刺激。研究了心房利钠因子（ANF）对克隆的人肾肿瘤（hKT）细胞和人肾肿瘤上皮细胞系（SK-NEP-1）的亲本细胞中环状鸟苷单磷酸（cGMP）和环状腺苷单磷酸（cAMP）积累的刺激作用。人ANF-(99-126)（10⁻⁶M）以剂量依赖性方式刺激（p<0.001）细胞cGMP积累，从基础水平0.26±0.04增加到3.73±0.81 pmol/mg蛋白质/5分钟（平均值±标准误，n = 13）。ANF对cGMP积累的刺激是特异性的，因为高浓度（10⁻⁶M）的心房肽I[大鼠ANF-(103-123)]、血管紧张素II、精氨酸加压素和氨氯地平（10⁻⁴M）不会增加基础cGMP。氨氯地平（10⁻⁴M）增强（p<0.01，n = 6）了ANF对cGMP积累的刺激作用（1.24±0.39 pmol/mg蛋白质/5分钟），特别是在低剂量ANF（10⁻¹⁰M）时，此时无氨氯地平的ANF刺激（0.34±0.08 pmol/mg蛋白质/5分钟）与cGMP积累的基础水平（0.19±0.02 pmol/mg蛋白质/5分钟）几乎无法区分。用百日咳毒素预孵育细胞可减弱ANF的刺激作用（1.59±0.07 pmol/mg蛋白质/5分钟降至0.75±0.06 pmol/mg蛋白质/5分钟，p<0.01，n = 6），但霍乱毒素则无此作用。ANF（4.56±0.93 pmol/mg蛋白质/五分钟，n = 8）对hKT细胞中的cAMP积累（4.32±0.98 pmol/mg蛋白质/5分钟）无影响。这是关于ANF反应性人肾细胞系的首次报道，其应用应有助于研究ANF-受体相互作用。
