Brameld J M, Broughton Pipkin F, Symonds E M
Department of Obstetrics and Gynaecology, University Hospital, Queen's Medical Centre, Nottingham.
J Endocrinol. 1990 Dec;127(3):513-21. doi: 10.1677/joe.0.1270513.
The renal and genital tracts share a common embryological origin; it is thus not surprising that tissues from both can synthesize renin. Preliminary studies showed extremely high concentrations of renin in follicular fluid (FRC) following ovarian stimulation for in-vitro fertilization. This necessitated complete revalidation of the renin assays and showed that data obtained using commercial kits were invalid. An assay protocol was developed using a 1:2 dilution of follicular fluid taken into EDTA (0.3 mol/l) and o-phenanthroline (0.05 mol/l). The assay was performed at pH 7.5 in the presence of excess exogenous (sheep) renin substrate, with incubation periods of 5, 10 and 15 min at 37 degrees C. This protocol resulted in the linear generation of angiotensin I (AI). Activation of inactive renin was performed using eightfold more trypsin than was required for plasma samples. Follicular renin substrate concentrations (FRS) were measured using the same assay methodology as used for measurement of plasma renin substrate concentrations (PRS). Storage of samples at -18 degrees C for up to 2 months was found not to affect the FRC, although repeated freeze-thaw cycles did. FRC and plasma renin concentrations (PRC) were very similar in 25 unstimulated control women, studied in the follicular phase of the menstrual cycle. Trypsin activation increased follicular total renin concentration (FTRC) more than plasma total renin concentration (PTRC) (P less than 0.0001). FRS was slightly higher than PRS (P less than 0.02). Ovarian stimulation with clomiphene citrate (CC; six women) was without effect on these parameters.(ABSTRACT TRUNCATED AT 250 WORDS)
肾脏和生殖道有着共同的胚胎学起源；因此，来自这两者的组织都能合成肾素也就不足为奇了。初步研究表明，在为体外受精而进行卵巢刺激后，卵泡液（FRC）中的肾素浓度极高。这使得肾素检测必须完全重新验证，并表明使用商业试剂盒获得的数据是无效的。开发了一种检测方案，将卵泡液以1:2的比例稀释到含有乙二胺四乙酸（0.3 mol/l）和邻菲罗啉（0.05 mol/l）的溶液中。检测在pH 7.5的条件下进行，存在过量的外源性（绵羊）肾素底物，在37摄氏度下孵育5、10和15分钟。该方案导致血管紧张素I（AI）呈线性生成。使用比血浆样本所需量多八倍的胰蛋白酶来激活无活性肾素。卵泡肾素底物浓度（FRS）的测量方法与血浆肾素底物浓度（PRS）的测量方法相同。发现样本在-18摄氏度下储存长达2个月不会影响FRC，不过反复冻融循环会有影响。在月经周期卵泡期对25名未受刺激的对照女性进行研究，发现FRC和血浆肾素浓度（PRC）非常相似。胰蛋白酶激活使卵泡总肾素浓度（FTRC）的增加幅度大于血浆总肾素浓度（PTRC）（P小于0.0001）。FRS略高于PRS（P小于0.02）。用枸橼酸氯米芬（CC；六名女性）进行卵巢刺激对这些参数没有影响。（摘要截断于250字）
