Renin-like Activity Research Articles

The Role of Extrarenal Renin in Goldblatt Hypertension

Renin-like activity can be demonstrated in arterial extracts from normal rats and from rats with Goldblatt hypertension. We have found no evidence, however, for elevated arterial renin levels in relation to plasma renin activity. Studies of the reversal of renovascular hypertension indicate that the fall in blood pressure produced by renal artery deconstriction is not renin-dependent. However, molecular biological techniques indicate that extrarenal renin gene expression is altered in some tissues, such as the adrenal gland in Goldblatt hypertension, and it is possible therefore that extrarenal renin synthesis is important in blood pressure control. This has to be distinguished from renin derived from plasma uptake.

Production of angiotensinogen and renin-like activity by rabbit proximal tubular cells in culture

Recent studies suggest the presence of local angiotensin generating system in the kidney. By using in situ hybridization technique, mRNA for angiotensinogen has been shown to be present in the proximal tubule. In the present study, we have attempted to examine the production of angiotensinogen and renin-like activity by the proximal convoluted (PCT) and straight (PST) tubular cells. PCT and PST cells were obtained from microdissected rabbit proximal tubules and cultured in vitro. Angiotensinogen and renin-like activity were quantitated in culture media and cell lysates. It was found that PCT culture medium contained both angiotensinogen and renin-like activity, whereas only angiotensinogen was detected in PST culture medium. Support for de novo synthesis is provided by the observation that both angiotensinogen and renin-like activity in PCT culture medium increased in a time-dependent and hormone-sensitive manner in defined serum-free medium. These results thus demonstrate the actual production of angiotensinogen and renin-like activity by proximal tubular cells, and indicate that these locally synthesized components may contribute to the regulation of angiotensin generation in renal proximal tubule.

Increased plasma prorenin but not renin after bilateral ureteral ligation in dogs

Plasma prorenin is normally higher than renin and usually changes in response to the same stimuli. In dogs, plasma prorenin and renin disappear after bilateral nephrectomy, indicating that both are of renal origin. It has been proposed that prorenin may mediate tissue renin systems via its reversible intrinsic renin-like activity. The renin-angiotensin system has been implicated in the changes in renal function that occur with bilateral ureteral obstruction, but plasma prorenin has not been investigated. We therefore studied the effect of 48-hour bilateral obstruction on plasma prorenin in two groups of dogs: one was volume expanded (N = 5), while the other group (N = 6) was euvolemic. Plasma prorenin concentration increased fourfold in both groups, angiotensinogen increased twofold, while plasma renin activity was unchanged. Following release of obstruction, plasma renin fell slightly while prorenin and angiotensinogen remained elevated. There was a positive relationship between plasma prorenin and renin before (r = 0.63, P less than 0.05) and after (r = 0.76, P less than 0.01) obstruction. Post-obstruction, ERPF and GFR were subnormal but filtration fraction was maintained; the higher the ERPF and GFR the higher the plasma prorenin post-obstruction (r = 0.83, P less than 0.01 and r = 0.77, P less respectively; N = 11). These results show that impairment of renal function during bilateral obstruction is associated with an increase in plasma prorenin but not renin. Nonetheless, there is a positive relationship between plasma prorenin and renin both pre- and post-obstruction. Thus, preferential impairment of clearance of prorenin relative to renin may occur during bilateral obstruction.(ABSTRACT TRUNCATED AT 250 WORDS)

Renin-angiotensin system in antarctic fishes

1. 1. The presence of a renin-angiotensin-like system has been investigated in the Antarctic fishes Chionodraco hamatus (Fam. Channichthydae) and Pagothenia ( Trematomus) bernacchii (Fam. Notothenidae). 2. 2. A renin-like activity is present in plasma and kidney of both the white blooded ( Chionodraco) and the red blooded ( Pagothenia) species. 3. 3. An angiotensin converting enzyme-like activity has been demonstrated in plasma, gills and kidneys of both species. The activity is inhibited by high temperature. 4. 4. From our data a renin-angiotensin-like system is present in the Antarctic fishes studied but the cascade of enzymes is active only at low temperatures.

Effect of Angiotensin-Converting Enzyme Inhibition on Human Tissue Renin

This investigation was performed to assess the effect of treatment with an angiotensin-converting enzyme (ACE) inhibitor, ramipril, on human tissue renin levels. A total of 41 patients were studied (32 males and 9 females, aged 48-71 years). Twenty-two of these patients received up to 5 days treatment with the ACE inhibitor ramipril (5 mg p.o.) prior to surgery, the remaining 19 patients served as controls (no ramipril treatment). Under surgery vascular tissues were removed and renin-like activities measured in carotid and renal arteries. Plasma renin and ACE activity were determined before and after surgical intervention. Vascular renin values were two- to threefold higher in ACE-treated hypertensive patients compared to normotensive and untreated hypertensive controls (irrespective of concomitant atherosclerotic lesions). There were no differences in vascular renin observed between primary (essential) and secondary hypertension. In the hypertensive patients treated with ramipril, blood pressures were markedly (but not significantly) lowered in 12 patients. A similar pattern was observed for treatment-induced changes in plasma renin (increased) and ACE activity (decreased). The findings of this study demonstrate that treatment with an ACE inhibitor, such as ramipril, evokes changes in extrarenal tissues similar to those demonstrable in plasma. Conceivably, the blood pressure-lowering effect of ACE inhibition on human tissue is partially induced at the vascular level.

Vascular Renin and Hypertension: Uptake Versus Synthesis

Conventional radioimmunoassay techniques demonstrated in the aortic wall a renin-like activity which is derived from plasma but has a longer half-life than plasma renin. Blood pressure elevation after renin injection into nephrectomized rats correlates better with aortic renin than with plasma renin. Vascular and other extrarenal tissue can also synthesize renin. Using a ribonuclease protection technique for the detection of renin messenger RNA we have been able to demonstrate that a wide variety of extrarenal tissues contain the renin message. In at least two of these, the brain and the liver, renin messenger RNA levels are unaffected by changes in dietary salt or by changes in systemic blood pressure. Functional studies using isolated human resistance vessels also demonstrate the presence of renin-like activity by a contractile response to added renin substrate. It is suggested that extrarenal tissues therefore contain renin-like activity derived both from uptake and from local synthesis. These systems may be regulated in different ways and may carry out different functions.

Effects of angiotensin II, ACTH, and KCl on the adrenal renin-angiotensin system in the rat

Angiotensin II, ACTH and potassium chloride were administered to rats for 6 days and the effects on adrenal renin-like activity and adrenal angiotensin II/III immunoreactivity were investigated. Rats infused with angiotensin II (140 pmol/min) either ip or sc showed increases in adrenal angiotensin II/III immunoreactivity (p less than 0.05) and plasma aldosterone concentration (p less than 0.05), but no change in adrenal renin-like activity. Captopril treatment of angiotensin II-infused rats caused a slight decrease in angiotensin II/III immunoreactivity which did not reach statistical significance. In contrast, rats treated with ACTH (Cortrosyn-Z, 3 IU/day, sc) showed an increase in adrenal renin-like activity (p less than 0.01), but no significant change in adrenal angiotensin II/III immunoreactivity. Rats treated with KCl in drinking water showed increases (p less than 0.05) in adrenal renin-like activity, adrenal angiotensin II/III immunoreactivity, and plasma aldosterone. These results suggest that angiotensin II, ACTH and potassium, three major regulators of aldosterone secretion by the adrenal gland, have different effects on the adrenal renin-angiotensin system when administered in vivo.

The Renin-Angiotensin System in Different Stages of Spontaneous Hypertension in the Rat (S H R)

The present study analyzed the concentration of renin-like activity and angiotensinogen concentration (AoC) in different brain areas related to cardiovascular control in SHR and Wistar Kyoto (WKY) animals. Male rats of both strains were studied at 8, 16 and 30 weeks of age. The following brain areas were isolated: anterior, medial and posterior hypothalamus, septal area, periaqueductal gray (PG) and the remaining brain stem; nucleus tractus solitarius (NTS) and the remaining medulla oblongata. Plasma renin activity (PRA) and plasma and cerebrospinal fluid (CSF) AoC were determined. Renin-like concentration was higher in SHR than in WKY in the anterior hypothalamus, PG and NTS at different stages of hypertension development. AoC was also higher in some areas of the SHR brain during different periods. PRA, plasma and CSF angiotensinogen concentration showed significant differences between both strain of rats during the development of high blood pressure. Present data support the possibility that the central and peripheral renin-angiotensin system may participate in the maintenance of high blood pressure in the SHR animals.

Effects of renin inhibitors on the expression of kidney renin gene and tissue renin-like activity.

The effect of renin inhibitor ES-1005 on renin gene expression was investigated in sodium-depleted marmosets. The kidneys were removed after continuous infusion of ES-1005 (12 mg/kg per h) for 2 h. The relative amount of kidney renin messenger (m)RNA was measured by densitometric Northern blot analysis using an alpha-32P-labelled human renin complementary (c)DNA fragment as a hybridization probe. Plasma renin activity was completely inhibited by ES-1005. The level of kidney renin mRNA decreased significantly to about one-third of the normal control value. We also investigated the inhibitory potency of the renin inhibitor ES-6864 on the renin-like activity in dog tissues (adrenal glands, aorta and brainstem). ES-6864 inhibited the tissue renin-like activity with an IC50 of 10(-7) to 10(-8) mol/l in vitro. Renin inhibitors not only inhibit the activity of plasma and tissue renin, but also suppress the synthesis of renin in the kidney.

Extrarenal renin and blood pressure regulation. An alternative viewpoint.

There is increasing evidence that a major site of production of angiotensin I and II is peripheral tissue. Both angiotensin I and II are present in venous blood in amounts far too high to be explained by their generation in blood alone, given the extensive conversion of angiotensin I to angiotensin II and clearance of both peptides across peripheral tissues. Much indirect evidence supports this argument for angiotensin production in tissues, and indicates that tissue production of angiotensin plays an important role in regulation of blood pressure. Tissue renin may represent uptake from plasma and/or local synthesis, but despite the ease with which renin-like activity can be measured in tissues, its interpretation is problematic because of interference by nonrenin enzymes and inadvertent activation of inactive renin. Moreover, given that enzymes other than renin are able to liberate angiotensin I and angiotensin II from angiotensinogen, there is no obligatory role for renin in angiotensin production in tissues. Inasmuch as tissue production is the major source of plasma angiotensin, the fall in plasma angiotensin levels after bilateral nephrectomy indicates that kidney-derived renin is the major contributor to tissue angiotensin production. This argument is supported by evidence that vascular renin-like activity is kidney-derived, and plays a dominant role in angiotensin-dependent pressor mechanisms. Near-normal levels of inactive renin in plasma of anephric subjects indicates extrarenal synthesis of inactive renin, and renin mRNA has been identified in various nonrenal tissues. Whether these tissues are also able to process inactive renin to active renin, and its role in local angiotensin production and blood pressure regulation, are currently being investigated.

Distribution of Angiotensin II Receptors and Renin in the Mouse Fetus

To investigate the ontogeny of the renin-angiotensin system we studied the characteristics and location of angiotensin II (AII) receptors in mouse fetuses and examined sites of renin mRNA expression by in situ hybridization and Northern blot analysis. Autoradiographic analysis of the binding of 125I-[Sar1,Ala8]AII to slide-mounted frozen sections of 17-day-old DBA/2N mice revealed abundant AII receptors widely distributed throughout the body. High receptor density was found in primitive mesenchymal tissue under the epidermis and surrounding muscle and cartilage, in skeletal and smooth muscle, and in all layers of the adrenal cortex. Lower receptor density was seen in the kidney, liver, and lungs. The autoradiographic staining was abolished by incubation of the sections with excess unlabeled AII. Scatchard analysis of the binding of 125I-[Sar1,Ala8,]AII to membrane-rich fractions of eviscerated fetuses showed a single type of high affinity receptors with a Kd of 2.9 x 10(-9) M and a receptor concentration of 3300 fmol/mg protein. Localization of renin mRNA was analyzed by in situ hybridization using an antisense 35S-labeled riboprobe transcribed from a mouse renin2 cDNA clone. Hybridization to fetal tissue sections showed high intensity staining in the kidney and adrenal cortex. Northern blot analysis confirmed the high expression of renin mRNA in the fetal kidney. The presence of an active renin-angiotensin system in the fetus was confirmed by the demonstration of renin-like activity and bioactive AII in fetal extracts. The widespread distribution of AII receptors in the fetus, compared to the discrete localization to specialized tissues in the adult, may indicate a unique role for the peptide during development.

In vitro and in vivo inhibition of renin by a thiazolylalanyl cyclostatine derivative

A compound containing cyclostatine ES-6864 (N-[(2R)-3-morpholinocarbonyl-2-(1-naphthylmethyl)propionyl]-(4-th iazolyl)- L-alanyl-cyclostatine-(2-morpholinoethyl)amide) was found to be a competitive inhibitor of human renin with an inhibitory constant (Ki) value of 7.3 x 10(-9) M. The compound was also potent against monkey renin but was less effective against renins from pig, goat, dog, rabbit and rat. ES-6864 did not inhibit cathepsin D, pepsin, urinary kallikrein, angiotensin converting enzyme, trypsin and chymotrypsin at a concentration of 10(-5) M. ES-6864 also inhibited the tissue renin-like activity from dog tissues with IC50 values of 10(-7)-10(-8) M in vitro. Oral administration of ES-6864 at 30 mg/kg to conscious, sodium-depleted marmosets produced a significant blood pressure reduction and a significant inhibition of plasma renin activity, which persisted for 6 hours. Plasma concentration of ES-6864 reached a maximum of 1.2 micrograms/ml at 1 hour after an oral administration. Oral administration of ES-6864 to hog renin-infused rats produced dose-related decreases in blood pressure. The results demonstrate that ES-6864 is an orally active renin inhibitor with high potency and specificity for human renin. Thus, ES-6864 is a candidate compound for development of renin inhibitors that can be used clinically.

80 Effects of chronic administration of a renin inhibitor on renin-like activity in marmoset tissues

80 Effects of chronic administration of a renin inhibitor on renin-like activity in marmoset tissues

Similarity between active and trypsin-activated inactive renin in dog plasma by means of renin inhibition: the dog as an animal model for studies of inactive renin.

A method for trypsin-activation of dog plasma inactive renin is described. Liquid phase trypsin (final concentration 6.7 mg/ml) was used and the reaction was stopped after 2 min at 4 degrees C by soybean trypsin inhibitor (13 mg/ml). Renin was measured as angiotensin I (Ang I) generation in trypsin-treated and untreated plasma using the antibody-trapping method, in the presence of excess ox renin substrate. The renin-like activity after trypsin was indeed due to renin, since Ang I generation in dog plasma before and after trypsin treatment was completely inhibited by H-77 at 10(-6) mol/l, and the two IC50 values were very similar (2.7 +/- 0.7 and 2.9 +/- 0.7 at 10(-8) mol/l, respectively). Dog plasma inactive renin was effectively separated from active renin by chromatography on Affigel Blue. Like human prorenin, dog plasma inactive renin rose in response to sodium depletion (furosemide 5 mg/kg, i.v.) followed by a low-salt diet (1 mmol Na+/day) for 4 days, (from 29.6 +/- 8 to 162 +/- 22 microU/ml; P less than 0.01, n = 10). Active renin also increased as expected. Intravenous captopril (6 mg/kg per h), for 3 h, led to a sharp increase in dog plasma active renin (from 53 +/- 8 to 360 +/- 60 microU/ml; P less than 0.01, n = 6), whereas inactive renin remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Vascular and adrenal reninlike activity in chronically diabetic rats.

The aim of this work was to investigate, in an experimental model of diabetes mellitus, the levels of renin activity in vascular and adrenal tissues and their relationship to several circulating renin-angiotensin system components. Rats with chronic (12 weeks) streptozocin-induced diabetes showed a significant decrease in plasma renin activity (PRA), plasma renin concentration, and plasma aldosterone. However, plasma trypsin activatable inactive renin concentration was increased (11.65 +/- 1.40 vs 6.73 +/- 0.57 ng angiotensin I/ml/hr; p less than 0.001), as were aortic reninlike activity (p less than 0.001) and adrenal renin, both in the zona glomerulosa (p less than 0.01) and the fascicular-reticular-medullary portion (p less than 0.001) with respect to an age-matched control group. After bilateral nephrectomy, plasma renin-angiotensin system components (PRA and plasma active and inactive renin concentrations) as well as aortic and fascicular-reticular-medullary renin activity significantly decreased in both control and diabetic rats. However, glomerular renin activity increased in control nephrectomized rats to the levels observed in diabetic animals but did not change in diabetic nephrectomized rats. The parallel changes of aortic and fascicular-reticular-medullary renin activity and plasma inactive renin concentration in diabetes and nephrectomy suggest an interdependent relationship, whereas the increase of glomerular renin activity in diabetic and nephrectomized animals, both with low levels of PRA, suggests the existence of a local autonomic renin-angiotensin system regulated by plasma feedback. Tissue renin-angiotensin system alterations in diabetes could mean that a pathogenic factor is involved in long-term diabetic complications or that only a compensatory physiological process is at work.

The ovarian renin-angiotensin system: Renin-like activity and angiotensin II/III immunoreactivity in gonadotropin-stimulated and unstimulated human follicular fluid

The ovarian renin-angiotensin system: Renin-like activity and angiotensin II/III immunoreactivity in gonadotropin-stimulated and unstimulated human follicular fluid

Human ovarian theca cells are a source of renin.

Human ovarian follicular fluid contains renin-like activity. In normal women, circulating levels of prorenin, the biosynthetic precursor of renin (EC 3.4.23.15), change in parallel with changes in progesterone during the menstrual cycle. Therefore, the ovary has been implicated as a source of plasma prorenin. In the present studies, we report the finding of high concentrations of prorenin in human ovarian follicular fluid (3000 ng.ml-1.hr-1 vs. 10-40 ng.ml-1.hr-1 in normal human plasma) obtained from follicles of women prepared for in vitro fertilization. The inactive renin-like enzyme was identified as prorenin by its activation characteristics, its molecular weight of 47,000, which is the same as that for recombinant prorenin, and its cross-reactivity with human renal renin antibodies. Culture of isolated human theca cells and isolated granulosa cells indicated that prorenin is secreted by theca cells but not by granulosa cells. Prorenin production by theca cells peaked during the first 10 days of culture and gradually decreased by 17 days. Active renin levels were 10% or less of the prorenin levels. Prorenin was barely detectable in medium from granulosa cells cultured for 24 days. Immunohistochemical staining of human ovaries (n = 5) with anti-human renin antibody demonstrated the presence of renin primarily in theca cells. These studies suggest that the theca cell is the source of the large quantities of prorenin in human ovarian follicular fluid.

Renin-angiotensin system.

Renin, a protease that cleaves the prohormone angiotensinogen thereby releasing angiotensin I, plays a major role in regulating blood pressure and electrolyte balance. Although renin is synthesized mainly in the cortical cells of the kidney, the presence of renin or reninlike activity has been recognized in various extrarenal tissues of humans and animals, including such reproductive organs as LH-containing cells of the anterior pituitary, placentral trophoblasts, and Leydig cells of the male gonad. It has also been learned that the renin in these extrarenal tissues generates angiotensins locally, which may interact with angiotensin II receptors in a paracrine or autocrine manner. However, the precise roles of the renin-angiotensin system in extrarenal tissue are not yet understood. We demonstrated the renin-angiotensin system in human testis by measuring the plasma renin activity (PRA), plasma angiotensin II (PAII), and testosterone in the internal spermatic vein (ISV) simultaneously after treatment with hCG, and elucidated the relationship between the renin-angiotensin system and sex steroids from Leydig cells of the male gonad.

Osmotic stress, plasma renin activity, and spermatogenesis in Vipera aspis

Circulating electrolytes (Na +, K +), plasma renin-like activity, testosterone, and testis morphology were investigated in early summer during the spermatogenic progressive phase in Vipera aspis subjected to sodium loading and sodium depletion. After sodium loading, plasma sodium and plasma testosterone levels were significantly elevated compared with those of controls, while plasma renin-like activity was depressed, spermiogenesis was increased, the epithelium lining the epididymis was very thick, and the Leydig cells were hypertrophied. After sodium depletion, plasma sodium and plasma testosterone levels were significantly depressed and plasma renin-like activity was significantly elevated. Spermiogenesis seemed to be slightly regressed: the epithelium lining the epididymis was very thin, and the lumen was devoid of spermatozoa. The Leydig cells were hardly visible. All the data strongly suggest that osmotic stress affects gonadal activity in the snake. V. aspis.

Inhibition of renin-like activity in marmoset tissues by the renin inhibitor CGP 29 287.

This study investigated the contribution of tissue renin-like activity to the acute hypotensive response induced by the potent and specific inhibitor of primate renin, CGP 29 287. Furosemide-pretreated marmosets (5mg/kg, i.m.) were killed 30 min after bolus injection of CGP 29 287 (1 mg/kg, i.v., n = 4). At this time, plasma renin activity is completely inhibited and the hypotensive response is maximal. Control marmosets received vehicle (0.9% saline, 1 ml/kg, i.v.). Renin-like activity was measured by the rate of angiotensin I (ANG I) formation after incubation of tissue homogenates with sheep angiotensinogen at pH 6.0 and 7.4. Activity was detected in the brain, heart, aorta and kidney of CGP 29 287-treated and control marmosets, and there were no significant differences in the values between the two groups. Addition of CGP 29 287 (5 X 10(-6) mol/l) to tissue homogenates in vitro inhibited ANG I formation (greater than 80%). These results indicate that CGP 29 287 is an inhibitor of tissue renin-like activity in vitro, but does not inhibit activity in these tissues after acute administration in vivo. Thus, the acute hypotensive response to CGP 29 287 appears to be due to inhibition of circulating renin only.