Shimamoto K, Tanaka S, Nakao T, Ando T, Nakahashi Y, Sakuma M, Miyahara M
Jpn Circ J. 1979 Mar;43(3):147-52. doi: 10.1253/jcj.43.147.
We have established a simplified assay system for the measurement of urinary kallikrein activity by utilizing the sensitive and specific radioimmunoassay system of kinins previously reported from our laboratory. Kinins were generated by incubating urine samples (50 microliter) with kininogen (1500 ng) in the presence of kininase inhibitors, and the generated kinins were measured by radioimmunoassay. Since the cross reactivity of kininogen in the kinin radioimmunoassay system was not recognized at dose up to 1.0 microgram, the amount of untreated kininogen in the radioimmunoassay samples did not interfere with the measurement of kinins. This eliminated the necessity for a kininogen extraction procedure. A good linear correlation (r = 0.939, p less than 0.001) was observed between the urinary kallikrein activity determined by this assay system (kininogenase activity) and that by esterolytic acitvity. Urinary kallikrein activity was 3.3 +/- 0.9 microgram/min/24 hour urine (mean +/- SEM), 1.4 +/- 0.4 microgram/min/24 hour urine and 0.25 +/- 0.06 microgram/min/24 hour urine in 6 normal subjects, 7 patients with non-complicated essential hypertension and 4 patients with chronic renal failure, respectively. Thus, urinary kallikrein activity was significantly lower in the patients with essential hypertension (p less than 0.05) and the patients with chronic renal failure (p less than 0.01) than in the normal subjects.
我们利用本实验室先前报道的灵敏且特异的激肽放射免疫分析系统,建立了一种用于测量尿激肽释放酶活性的简化检测系统。通过在激肽酶抑制剂存在的情况下,将尿样(50微升)与激肽原(1500纳克)一起孵育来生成激肽,然后用放射免疫分析法测定生成的激肽。由于在剂量高达1.0微克时,激肽放射免疫分析系统中未识别出激肽原的交叉反应性,放射免疫分析样品中未处理的激肽原量不会干扰激肽的测量。这就消除了激肽原提取步骤的必要性。该检测系统测定的尿激肽释放酶活性(激肽原酶活性)与酯解活性之间观察到良好的线性相关性(r = 0.939,p < 0.001)。6名正常受试者、7名无并发症的原发性高血压患者和4名慢性肾衰竭患者的尿激肽释放酶活性分别为3.3±0.9微克/分钟/24小时尿(平均值±标准误)、1.4±0.4微克/分钟/24小时尿和0.25±0.06微克/分钟/24小时尿。因此,原发性高血压患者(p < 0.05)和慢性肾衰竭患者(p < 0.01)的尿激肽释放酶活性显著低于正常受试者。