Higaki J, Ogihara T, Imai N, Kumahara Y, Hontani S, Nishiura M, Ogawa H, Hirose S, Murakami K
J Lab Clin Med. 1984 Dec;104(6):947-54.
We developed new sensitive direct radioimmunoassay for human plasma renin. Renin was purified from Haas' preparation utilizing a pepstatin-C6-Sepharose affinity chromatography. Antiserum, prepared by immunizing rabbits with the purified renin, was used for the direct radioimmunoassay at a final dilution of 1:30,000. The antibody was specific for human renal and plasma renin, but did not cross-react with cathepsin D, trypsin, or renins of mouse, dog, and rat. Radioimmunoassay was performed by the double antibody technique using the delayed tracer addition method. In this method, a standard curve was obtained over a range from 0.2 to 8.0 ng/ml. The values from our assay correlated well with total renin activity measured as the generation rate of angiotensin I after trypsin activation (r = 0.78, p less than 0.01), but correlated weakly with active renin activity. This finding disclosed that both active and inactive renin were detected by this method. In normal participants, plasma renin concentration determined by direct radioimmunoassay was increased by standing and furosemide injection. The plasma renin concentration determined by direct radioimmunoassay of patients with essential hypertension (0.7 to 1.7 ng/ml) was not significantly different from values in normal controls (0.8 to 1.9 ng/ml). The values were higher in patients with renovascular hypertension (1.6 to 2.7 ng/ml), malignant hypertension (2.8 to 3.4 ng/ml) and Bartter's syndrome (1.8 to 2.5 ng/ml), but lower in patients with primary aldosteronism (0.4 to 0.8 ng/ml) than in normal controls. This newly developed radioimmunoassay for human renin was sensitive enough to estimate the levels of renin in plasma of patients with low renin hypertension. It provides a new tool for the understanding of the renin-angiotensin system under various clinical conditions.
我们开发了一种用于检测人血浆肾素的新型灵敏直接放射免疫分析法。利用胃蛋白酶抑制剂 - C6 - 琼脂糖亲和色谱法从哈斯制剂中纯化肾素。用纯化后的肾素免疫兔子制备的抗血清用于直接放射免疫分析，最终稀释度为1:30,000。该抗体对人肾和血浆肾素具有特异性，但与组织蛋白酶D、胰蛋白酶以及小鼠、狗和大鼠的肾素无交叉反应。放射免疫分析采用双抗体技术并使用延迟加样法。用此方法得到了0.2至8.0 ng/ml范围内的标准曲线。我们分析得到的值与经胰蛋白酶激活后以血管紧张素I生成速率测量的总肾素活性相关性良好（r = 0.78，p小于0.01），但与活性肾素活性相关性较弱。这一发现表明该方法能检测到活性和非活性肾素。在正常受试者中，通过直接放射免疫分析测定的血浆肾素浓度在站立和注射速尿后会升高。原发性高血压患者通过直接放射免疫分析测定的血浆肾素浓度（0.7至1.7 ng/ml）与正常对照组的值（0.8至1.9 ng/ml）无显著差异。肾血管性高血压患者（1.6至2.7 ng/ml）、恶性高血压患者（2.8至3.4 ng/ml）和巴特综合征患者（1.8至2.5 ng/ml）的值较高，但原发性醛固酮增多症患者（0.4至0.8 ng/ml）的值低于正常对照组。这种新开发的人肾素放射免疫分析法灵敏度足以估计低肾素性高血压患者血浆中的肾素水平。它为在各种临床情况下理解肾素 - 血管紧张素系统提供了一种新工具。
