Role of changes in renal hemodynamics and P-450 metabolites of arachidonic acid in the reversal of one-kidney, one clip hypertension.

OBJECTIVE: To examine the role of changes in renal hemodynamics and P-450 metabolites of arachidonic acid in the reversal of one-kidney, one clip (1-K,1C) hypertension in rats. DESIGN: The stimulus for the release of an antihypertensive lipid from the kidney is not known. This study examined whether cortical or papillary blood flow is altered after removal of the clip from the renal artery of 1-K,1C hypertensive rats, and the effects of blockade of the renal metabolism of arachidonic acid by P-450 with 17-octadecynoic acid (17-ODYA) on the fall in blood pressure. METHODS: Cortical and medullary blood flows were measured using laser-Doppler flowmetry. 17-ODYA (33 nmol/min) was infused directly into the renal artery to examine the effect of inhibition of renal P-450 activity on reversal of 1-K,1C hypertension. The renal metabolism of arachidonic acid in control and in 1-K,1C hypertensive rats was assessed by incubating microsomes with [14C]-arachidonic acid, the metabolites formed being measured using reverse-phase high-performance liquid chromatography. The antihypertensive effects of these P-450 metabolites of arachidonic acid were compared with those of medullipin I after intravenous administration in conscious spontaneously hypertensive rats (SHR). RESULTS: Cortical and papillary blood flow increased significantly and arterial pressure fell after unclipping the renal artery in the 1-K,1C hypertensive rats. 17-ODYA prevented the fall in blood pressure after unclipping. The production of epoxy- and dihydroxy-eicosatrienoic acids was elevated in microsomes prepared from the renal cortex of the 1-K,1C hypertensive rats. However, intravenous administration of these metabolites did not mimic the effect of medullipin I to lower arterial pressure in SHR. CONCLUSION: Elevations in renal cortical or papillary blood flow, or both, may stimulate the release of a P-450-derived antihypertensive lipid from the kidney after unclipping of the renal artery in 1-K,1C hypertensive rats. However, it is unlikely that this substance is a P-450 metabolite of arachidonic acid.

Purification of class I medullipins from the venous effluent of isolated normal kidneys perfused under high pressure with saline.

Medullipin I (Med I) is a vasodepressor prohormone which is continuously elaborated into the renal venous effluent (RVE) of isolated rat kidneys perfused under high pressure. We have improved the yield of Med I by substituting saline for the albumin perfusate previously reported; and considerably improved refinement by directly fractionating the crude lipid extract of the RVE with high pressure liquid chromatography. The results show that Med I, as defined by previous physiologic and pharmacologic criteria, is not a single molecule. The 3 Class I medullipins described here are distinguished by subtle or overt differences in polarity and biologic activity.

Secretion of medullipin I by the kidney involves the cytochrome P-450 enzyme system.

OBJECTIVE: To determine whether secretion of medullipin I by the kidney is dependent on the cytochrome P-450 enzyme system in Sprague-Dawley and spontaneously hypertensive rats (SHR). METHODS: Isolated kidneys from Sprague-Dawley rats were perfused with 5% human albumin gassed with 95% O2 and 5% CO2 at 185 mmHg. The resultant renal venous effluent was tested in the SHR for medullipin I-type vasodepressor activity. The kidneys were then treated with ketoconazole, and inhibitor of the cytochrome P-450 enzyme system. After rinsing with 50 ml saline, the last 10 ml of which was saved for a control test (see below), the kidneys were reperfused with 50 ml human albumin and the resultant renal venous effluent was tested for vasodepressor activity. One milliliter of the saline rinse was administered to the SHR and the preketoconazole renal venous effluent was administered after 15 min. The medullipin I-type vasodepression occurred. Thus, inhibition of vasodepression after ketoconazole treatment was not due to residual ketoconazole in the post-treatment renal venous effluent. RESULTS: Treatment of isolated kidneys with ketoconazole prevented secretion of medullipin I which had been induced by 5% human albumin. CONCLUSION: The cytochrome P-450 enzyme system is involved in two major metabolic steps of the medullipin system: synthesis of medullipin I by the kidney and conversion of medullipin I to medullipin II by the liver as shown previously.

Renal vasodepressor mechanisms: the medullipin system.

Renal vasodepressor hormone: Medullipin I is the renomedullary vasodepressor hormone secreted by the renomedullary interstitial cells of the renal papilla. It is conveyed to the liver where it is converted to its active form, medullipin II. Medullipin II is a vasodilator that suppresses sympathetic tone and causes diuresis and natriuresis. Its actions are opposite to those of angiotensin II. These are feedback control systems. The secretion and conversion of medullipin is related to the cytochrome P-450 dependent enzyme system of kidney and liver. Deficiency of medullipin: A deficiency of medullipin is considered to contribute to the pathogenesis of various hypertensive states. There are three known causes for such a deficiency, (1) removal of renomedullary interstitial cells by bilateral nephrectomy, renal surgical papillectomy, chemical papillectomy, papillary atrophy or necrosis; (2) decrease in number and damage to renomedullary interstitial cells in accelerated experimental hypertension and malignant hypertension of humans; and (3) dysfunction of renomedullary interstitial cells as mediated by angiotensin II, by resetting of the effect of increased renal artery perfusion pressure, by stimulation of the renal sympathetic nerve, by inhibition of nitric oxide synthesis and possibly by inhibition of cyclo-oxygenase. Secretion of Medullipin I: The main factor influencing secretion of medullipin I by the kidney appears to be the renal artery perfusion pressure. Elevation of this pressure is attenuated by the presence of medullipin I in the renal venous effluent. Lowering the pressure below normal shuts off this secretion. This is opposite to the effects of perfusion pressure on renin secretion, as elevation shuts off renin secretion while depression turns it on.(ABSTRACT TRUNCATED AT 250 WORDS)

Lipomedullipinoma: a source of hypermedullipinemia.

Previously we reported a case of persistent hypotension associated with hypermedullipinemia (Blood Pressure 1992; 1:138-148). The hypermedullipinemia appeared to result from the autonomous secretion of medullipin I (Med I) by renomedullary interstitial cells (RIC's) in the patient's remaining endstage kidney. The patient subsequently died. At autopsy, the remaining kidney contained a yellow mass (1 x 1 x 0.5 cm) consisting of adipocytes and RIC's, termed a lipomedullipinoma. This mass was extracted and chromatographed by procedures known to yield Med I. Med I was identified following these procedures. Renal tissue outside the yellow mass failed to yield Med I. It appears that the hypermedullipinemia of this case resulted from autonomous, hypersecretion of Med I by the lipomedullipinoma.

Persistent hypotension associated with hypermedullipinemia: a new syndrome.

A new syndrome is described in a patient with advanced renal insufficiency. This consists of severe and persistent hypotension causing weakness but associated with a clear mental status. Also present is evidence for decreased vascular reactivity. The hypotension was not orthostatic. The hypotension was associated with a circulating vasodepressor substance having the characteristics of medullipin 1. The medullipin appears to have been derived from the remaining right kidney. Hypotension existed despite the presence of major prohypertensive mechanisms, including an endstage kidney, hyperreninemia and hyperaldosteronemia. It is likely that hypotension due to hypermedullipinemia is an entity occurring in the human being.

Secretion of medullipin I by the kidney requires oxygen.

OBJECTIVE: To test the hypothesis that the secretion of medullipin I by the kidney involves an oxidative step. DESIGN: Medullipin I is secreted by kidney renomedullary interstitial cells and is converted to medullipin II by the liver. Medullipin I can be derived from the kidney in the renal venous effluent by perfusing normal rat kidneys with 95% O2- 5% CO2 at an elevated pressure (180 mmHg). To evaluate whether the secretion of medullipin I involves an oxidative step normal rat kidneys were perfused at an elevated pressure in the presence of O2, in the absence of O2 and after treatment of the kidneys with a powerful antioxidant. METHODS: Normal rat kidneys were perfused with 5% albumin bubbled with O2-CO2 at 180 mmHg. This was the control procedure for each of the three approaches. In approach (1), the kidneys were perfused with 5% albumin bubbled with N2. In approach (2), the kidneys were perfused with 'blood' treated with carbon monoxide. In approach (3), the kidneys were treated with the antioxidant butylated hydroxytoluene then perfused with 5% albumin bubbled with O2-CO2. Each perfusate was tested for medullipin I activity by rapid intravenous injection into the SHR. RESULTS: All three approaches, which exclude the action of molecular O2, prevented the secretion of medullipin I by the kidneys. CONCLUSION: The secretion of medullipin I by the kidneys involves an oxidative step.

Biologic differences between vasodilator prostaglandins and medullipin I.

Vasodepressor prostaglandins (PGs), PGE2, PGI2, and medullipin I (Med I) are synthesized in the kidney. These vasodilator substances are thought to be involved in the antihypertensive function of the kidney. At issue is whether there are biologic differences between the vasodilator PGs and Med I. Two separate studies have shown that Med I's vasodepressor action is inhibited by four procedures: mixing with Tween 20; treatment with n-butyl boronic acid; treatment of the assay animal with SKF 525A, an inhibitor of cytochrome P-450; and removing the liver from the circulation. These same procedures were applied to the vasodilator PGs. All four failed to inhibit the vasodepressor action of the PG's. It is concluded that Med I and vasodilator PGs of the kidney are separate and distinct biologic entities.

Medullipin system. Generation of medullipin II by isolated kidney-liver perfusion.

Perfusion of isolated normal rat kidneys with blood at elevated blood pressure (180-200 mm Hg) followed by perfusion of the renal perfusate through an isolated normal rat liver under venous pressure yielded in the plasma of the hepatic effluent a vasodepressor lipid having the chromatographic and biological characteristics of medullipin II. These findings support the view that medullipin II is the final product of the renohepatic axis of blood pressure control.

The medullipin system of blood pressure control.

The medullipin system of BP control has its cellular component in the renomedullary interstitial cells (RICs) of the renal papilla. The RICs secrete medullipin I which is conveyed to the liver to be activated into Med II. This activation appears to involve the cytochrome P-450 dependent enzyme system of the liver. Med II is a vasodilator that suppresses sympathetic tone, causes diuresis-natriuresis and has a suppressive effect on the central nervous system (CNS). As such, Med II has biologic actions the exact opposite of major actions of the renin-angiotensin system. Angiotensin II is a vasoconstrictor that facilitates sympathetic tone, causes salt water retention by the kidney and has stimulatory effect on the CNS. These are double feedback BP control systems. One elevates the BP and is prohypertensive; the other lowers BP and is antihypertensive. Unclipped (UC) the one-kidney, one-clip hypertensive rat has assisted greatly in elucidating the medullipin system. The support for these contentions are documented in this presentation.

Antihypertensive action of medullipin I given by mouth.

Perfusion of normal rat kidneys with 5% human albumin in a balanced salt solution bubbled with oxygen yielded medullipin I (Med I) in the renal venous effluent. The presence of Med I in the renal venous effluent has been established by thin-layer chromatography, by the type of vasodepressor effect when injected intravenously as a bolus into the hypertensive rat, by inhibition of the vasodepressor effect of the renal venous effluent by Tween 20 and SKF 525A (proadifen, inhibitor of cytochrome P-450), and by removal of the liver from the circulation (a procedure that inhibits extracted Med I). Med I so derived lowered blood pressure of spontaneously hypertensive rats when injected into the stomach by an indwelling tube or when given by mouth. The lowering of blood pressure was attended by no change in cardiac output and no change in heart rate. Med I given by mouth to the spontaneously hypertensive rat is a vasodilator that suppresses sympathetic tone, acting in the same way as Med I extracted from renal papillae and given intravenously. Importantly, the antihypertensive action was demonstrated in the spontaneously hypertensive rat, a model of hypertension considered to mimic idiopathic or essential hypertension of humans. Med I is a promising therapeutic agent for hypertension.

Secretion of medullipin I by isolated kidneys perfused under elevated pressure.

1. Medullipin I (Med I) is a hormone extracted from renal papillae and its renomedullary interstitial cells (RIC). Med I is stimulated by elevation of the renal artery perfusion pressure. 2. When isolated normal rat kidneys were perfused either with oxygenated blood or with 5% albumin bubbled with O2 at elevated perfusion pressures, Med I appeared to be secreted into the renal venous effluent (RVE). Addition of Tween 20, treatment of the assay rat with SKF 525A, inhibitor of cytochrome P-450 and removal of the liver from the systemic circulation prevented vasodepression of both the RVE and extracted Med I. The lipid in the RVE gave the same dose-response as extracted Med I. 3. Lowering the renal artery perfusion pressure below normal inhibited the secretion of Med I. As the perfusion pressure was elevated Med I secretion appeared to increase. 4. Previous observations and the present study support the view that the renin-angiotensin system and the Medullipin system are double feedback systems involved in blood pressure control.

Renal function in rats with angiotensin II-salt-induced hypertension: effect of thromboxane synthesis inhibition and receptor blockade.

This study was designed to assess the contribution of thromboxane (Tx) A2 to the pathogenesis of renal dysfunction in rats with angiotensin II (Ang II)-salt hypertension. Hypertension was induced in rats drinking 0.15 mol/l NaCl by infusion of Ang II (125 ng/min, intraperitoneally) for 12 days. Relative to values in water- and saline-drinking rats without Ang II infusion, rats with Ang II-salt hypertension exhibited increased renal vascular resistance, decreased renal blood flow, and increased renal excretion and glomerular synthesis of TxB2. Treatment with an inhibitor of TxA2 synthesis, UK 38,485, had no effect on renal function in normotensive and hypertensive rats. Similarly, the TxA2 and prostaglandin endoperoxide antagonist SQ 29,548 did not affect renal function in normotensive rats. In contrast, in rats with Ang II-salt hypertension of 12 days' duration, SQ 29,548 caused a reduction in renal vascular resistance, allowing for maintenance of renal blood flow in the face of an accompanying reduction in blood pressure. A comparable reduction in renal perfusion pressure, produced by constriction of the abdominal aorta above the renal arteries, was not accompanied by a reduction in renal vascular resistance in Ang II-salt hypertensive rats. Therefore, the SQ 29,548-induced lowering of renal vascular resistance is attributable not to renal blood flow autoregulation, but to blockade of the renal vasoconstrictor actions of TxA2 and/or prostaglandin endoperoxides. This interpretation implies that pressor eicosanoids contribute to increase renal vascular resistance in rats with severe Ang II-salt hypertension.

Renal and circulatory effects of medullipin I, as studied in the in-vivo cross-circulated isolated kidney and intact Wistar-Kyoto (WKY) rat.

The renal medulla harbours powerful humoral antihypertensive mechanisms, as earlier explored in unclipping experiments on renal hypertensive rats or in normotensive isolated kidneys cross-circulated at increased perfusion pressures from 'donor rats', in which renal function also seemed to be affected. Injection of the renomedullary factor medullipin I (Med I; formerly ANRL) mimics these haemodynamic responses, and Med I seems to be one of the most important mediators of the depressor effects. The present study was performed to analyse further the haemodynamic and, particularly, the renal effects of Med I, using anaesthetized intact WKY rats and constant-pressure perfused (90 mmHg) isolated WKY kidneys, cross-circulated by these intact 'donor' rats. Mean arterial pressure (MAP), heart rate (HR) and renal function were followed for one 30-min period before and two 30-min periods after injection of 1 mg Med I (M; n = 7) or an equal volume of saline as control (C; n = 13). In the intact 'donor' WKY, MAP and HR remained largely constant in C during the three periods, being 126 +/- 5, 125 +/- 5, and 120 +/- 5 mmHg, while MAP fell in the M group after Med I, from 121 +/- 5 to 107 +/- 7 and 107 +/- 5 mmHg (P less than 0.05), and also HR tended to decrease in M. Renal resistance (RR) fell while renal plasma flow (RPF) and glomerular filtration rate (GFR) increased significantly (P less than 0.05) after Med I in the M donor rats despite their MAP reduction. However, in the constant-pressure perfused, cross-circulated kidneys the RR, RPF and GFR changes were clearly more pronounced (P less than 0.01) and also diuresis, natriuresis, osmolar excretion and osmolar clearance increased significantly after Med I (P less than 0.01). In conclusion, the present results support the view that Med I not only has important and long-lasting depressor effects but also affects renal function in important ways, inducing vasodilatation and increasing GFR, RPF, diuresis and sodium-osmolar excretion.

Histopathology of severe renal vascular damage in blacks.

During the 1970s renal biopsies were obtained after blood pressure had been controlled in 41 black patients in Memphis who had severe hypertension plus excretory renal failure. An additional 13 binephrectomy specimens were also studied. This material yielded significant information on the state of the renal arteries--arterioles under these circumstances. Fibrinoid necrosis of the afferent glomerular arteriole and proliferative glomerulitis were not noted. Rather, the vascular lesion characterized by the accumulation of smooth muscle cells and mucopolysaccharide in the intima attended by a marked narrowing of the lumen was dominant. This lesion caused pronounced ischemia associated with obsolescence of glomeruli, atrophy, and fibrosis (end-stage kidney). This lesion has been renamed musculomucoid intimal hyperplasia as a result of changes revealed by electron microscopic and histochemical studies. Since this study the incidence of this severe vascular disease of the kidney in the same geographic area has been markedly reduced. There are a number of possible reasons for this change in incidence, but a major one appears to be improved treatment of hypertension and better compliance with antihypertensive therapy. Why such extreme changes occur in a subset of hypertensive blacks is not known. It is apparent that without improved antihypertensive treatment, this type of end-stage renal disease due to severe vascular damage will continue to be encountered.

Biologic contrasts between medullipin I and vasoactive glyceryl compounds.

Medullipin I causes a delayed onset depressor response when injected intravenously into rats. The glyceryl compounds selachyl alcohol (SA) and monoolein (MO) cause similar vasodepression. The neutral lipid 1-O-hexadecyl-2-acetyl-sn-glycerol (HAG) was suggested by Blank et al to be medullipin I (Med I, formerly ANRL). Biologic comparisons were made between Med I and various glyceryl compounds, including SA, MO, HAG, alkyl glyceryl ethers of phosphatidyl choline (termed APRL by us), diacylated SA, and the n-butyl boronic acid derivative of SA and MO. The n-butyl boronic acid derivative of Med I also was evaluated. The delay in onset of the depressor response to Med I was reduced by the injection of Med I into the portal vein; that of SA and MO was not. Med I, SA, and MO were activated by the liver, while APRL and HAG were not. Tween 20 inhibited Med I, SA, and MO, but not APRL and HAG. Proadifen (SKF 525A) inhibited Med I, but not SA and MO. The n-butyl boronic acid derivatives of SA, MO, and Med I were inactive. Med I, like SA and MO, appeared to have two hydroxyl groups in close proximity. It was concluded that Med I is neither HAG, APRL, SA, nor MO.

The renal antihypertensive endocrine function: its relation to cytochrome P-450.

Unclipping the Goldblatt hypertensive rat lowers the blood pressure by cells in the renal papilla, the renomedullary interstitial cells (RIC), secreting a hormone that is part of a vasodilator system. A vasodilator, termed medullipin I, can be extracted from the renal papilla. Medullipin I and the renal venous effluent following unclipping have identical biologic properties. Medullipin I appears to be the agent secreted by the kidney following unclipping. Both medullipin I and the renal venous effluent must traverse the liver to be active. Medullipin I is converted in the liver to its active form, medullipin II. The blood pressure-lowering effect of both medullipin I and the renal venous effluent after unclipping are blocked by SKF 525A, the inhibitor of cytochrome P-450. The relation of the kidney to the liver was tested using the rate of decline of the blood pressure after unclipping as an index of the endocrine antihypertensive function of the kidney--acceleration of the decline being considered as increased function, decrease of the decline as decreased function. Five compounds: BW755C, phenobarbital, ketoconazole, eicosatetraynoic acid (ETYA) and butylated hydroxytoluene (BHT), and two manipulations: uretero-caval anastomosis (UCA) and removal of the liver from the circulation were used followed by unclipping. BW755C, inhibitor of both cyclo-oxygenase and lipoxygenase, potentiated the antihypertensive function to a maximum. It is reasoned that inhibition of the first two pathways of arachidonic acid metabolism potentiates the third pathway, the cytochrome P-450 pathway.(ABSTRACT TRUNCATED AT 250 WORDS)