Local Kinin Research Articles

Role of Prostaglandins in the Cardiovascular Effects of Bradykinin and Angiotensin-Converting Enzyme Inhibitors

The cardiovascular effects of bradykinin require additional vasoactive mediators for a fully balanced response. This includes arachidonic acid (eicosatetraenoic acid) and its metabolites, the eicosanoids (prostaglandins, leukotrienes, thromboxanes, and others). Eicosanoid generation by bradykinin is started by binding of the peptide to specific B2 receptors at the plasma membrane. This initiates G-protein coupled stimulation of phospholipase C, IP3-induced increases in cytosolic Ca2+, and stimulation of protein kinase C. Arachidonic acid is liberated from membrane phospholipids primarily via Ca(2+)-induced stimulation of phospholipase A2 and converted into tissue-specific eicosanoids by enzymes in the vicinity. In vascular tissue, most of the available arachidonic acid is converted into vasodilator prostaglandins, i.e., prostacyclin (PGI2) and prostaglandin E2 (PGE2). These prostaglandins are involved in vasodilator actions of the kinins. There is also some evidence for generation of vasoconstrictor eicosanoids, such as thromboxane A2, under certain conditions. The biological significance of kinin-related prostaglandin formation becomes apparent after inhibition of kinin breakdown by ACE inhibitors. These compounds prevent generation of vasoconstrictor angiotensin II and stimulate endothelial eicosanoid formation via local kinin accumulation. There is evidence suggesting that kinin-induced prostaglandin generation contributes to anti-ischemic, inotropic, and blood pressure-lowering effects of the compounds. This also includes inhibition of polymorphonuclear leukocyte (PMN) accumulation in injured myocardial tissue, which is antagonized by PGI2-related pathways, stimulated by ACE inhibition and/or bradykinin.

Inhibitory Effects of Kinins on Angiotensin I Conversion in the Local Circulation

The inhibition of angiotensin converting enzyme (ACE) by kinins was studied using bradykinin (BK), lysyl-bradykinin (Lys-BK), [Hydroxyproline3]-bradykinin ([Hyp3]-BK) and [Hydroxyproline3]-lysyl-bradykinin ([Hyp3]-Lys-BK). The latter two are novel kinins recently identified in our laboratory. All the four kinins displayed competitive inhibition on the conversion of angiotensin I to angiotensin II by purified canine lung ACE. Inhibition constants (Ki) for the four kinins were estimated from Dixon's plot as follows-BK: 0.27 microM, Lys-BK: 0.57 microM, [Hyp3]-BK: 0.34 microM, and [Hyp3]-Lys-BK: 0.27 microM. In the rat hindlimb perfusion system, the kinins were demonstrated to partially inhibit angiotensin I conversion to angiotensin II by the vascular ACE. Taken together, these results suggest that angiotensin II formation by ACE in the vascular tissue is possibly inhibited by local kinins, especially after ACE inhibitor administration. This indirect action of kinins, coupled with its direct vasodilatory action, might indicate a cooperative participation in vasodilation.

Metabolic Effects of Kinins

In 1928, Frey and co-workers discovered kallikrein in human urine and described its prolonged hypotensive effect in the dog. Four years later, the same authors first reported a blood glucose-lowering effect of orally administered kallikrein in diabetic patients. However, the observed blood glucose-lowering effect of kallikrein appeared to fade with repeated administration, and therefore its possible metabolic role was not further investigated and fell into disregard. One decade ago, experimental data yielded indirect evidence that the regulation of local skeletal muscle blood flow and glucose uptake during work was mediated by proteolytically cleaved kinins. Further experiments demonstrated that in insulin-resistant states such as postoperative stress and type II diabetes, reduced muscular insulin sensitivity was increased and partly restored by continuous low-dose infusion of synthetic bradykinin. Recent work showing that tissue kallikrein is present in a number of different tissue sites, including skeletal muscle and our own observation of local kinin overflow after muscle work in healthy subjects, but not in type II diabetics, support the concept of a skeletal muscle kallikrein-kinin system (KKS) that is locally activated upon contraction. Moreover, in isolated perfused rat heart preparations, favorable effects of kinins on myocardial glucose uptake, oxidation, and glycolytic flux have been reported. Most interestingly, cardioprotective effects of kinins have been observed and attributed to improved energy and substrate metabolism in ischemic hearts. Taken together, these data gave rise to the concept that tissue KKS might be involved in the local modulation of skeletal muscle and myocardial tissue blood flow and substrate metabolism, and that activation of the KKS is defective in insulin-resistant states.

Local generation of kinins in working skeletal muscle tissue in man.

The effect of standardized isometric forearm work on circulating and local kinin concentrations was investigated in 12 healthy volunteers using the forearm catheter technique. Radioimmunological kinin determination in arterial blood and in the venous effluent of forearm muscle tissue was performed using a modification of Shimamoto's technique of blood sampling and kinin extraction. Under basal conditions, there was no arterio-venous difference of kinins. Throughout the whole experiment, arterial--reflecting systemically circulating--kinins did not change. In muscle venous blood, immunoreactive kinins were not significantly elevated during work, whereas a marked increase was detected in the recovery period (5.0 +/- 0.6 vs. 10.2 +/- 2.0 pmol/l; p less than 0.01). The data demonstrate, that kinins are locally generated in calculated amounts (32.7 +/- 8.4 fmol/(100 g x min) that are known to be sufficient to induce local vasodilatory and metabolic effects at the site of muscle contraction, but below the threshold for systemic cardiovascular actions.

Effect of angiotensin-converting enzyme inhibition on circulating and local kinin levels

Angiotensin-converting enzyme kininase II reduces bradykinin metabolism in vitro and in vivo. However, consistent changes in circulating bradykinin levels after the angiotensin-converting enzyme inhibitor captopril have not been reported. The kallikrein-kinin system has been suggested to be a local hormonal system concerned with regional blood flow, and hence circulating levels may not reflect local tissue levels of kinins. Anesthetized dogs given captopril had a significant increase in urinary kinin excretion without a change in circulating bradykinin levels or in urinary kallikrein. These changes in renal kinins were accompanied by a decrease in blood pressure and renal vasodilation. The hypotension and renal vasodilation produced by captopril were not attenuated either by pretreatment with the angiotension receptor antagonist Sar 1-lleu 8-angiotensin II or by reduction of endogenous prostaglandin production with indomethacin. Postischemic renal vasodilation after temporary renal artery occlusion was also associated with increased urinary kinin levels. These results demonstrate that captopril effectively inhibits renal angiotensinconverting enzyme and that the renal kallikrein-kinin system may play an important role in regulating the renal vasculature and may contribute to the renal hemodynamic effects of captopril. Many polypeptide hormone membrane receptors are self-regulated by endogenous tissue concentrations of the peptide hormone. Infusions of bradykinin into rats reduced specific bradykinin receptors. A similar decrease in bradykinin receptor numbers without change in receptor affinity was demonstrated after captopril administration. These results provide indirect evidence that angiotensin-converting enzyme/kininase inhibition by captopril increases local tissue concentration of kinins, which may contribute to the hypotensive effect.

A study of the plasma kinin-generating system in children with the minimal lesion, idiopathic nephrotic syndrome.

Although the precise etiologic incitant of the minimal lesion idiopathic nephrotic syndrome of childhood is not known, it is likely that a host mechanism mediates the permeability alterations of the glomerular capillary wall resulting in massive proteinuria. As a first step in examining the possibility that local kinin release may account for the proteinuria in this disorder, two parameters of the plasma kinin-generating system, plasma prekallikrein and kallikrein inhibitor, were assayed during 27 nephrotic episodes in 21 corticosteroid-responsive children. Plasma kallikrein was assayed by means of its esterase activity on a synthetic arginine ester substrate, N-alpha-tosyl-L-arginine methyl ester (TAMe), after activation of Hageman factor by kaolin. This activity, after subtraction of spontaneous arginine esterase activity (i.e., TAMe esterase activity measured in plasma not exposed to kaolin) is derived from prekallikrein. Plasma prekallikrein activity in 11 normal children was 99.6 +/- 2.9 mumol TAMe hydrolyzed/ml plasma/hr (mean +/- SEM). Kallikrein inhibitor was quantified in arbitrary units. Kallifrein inhibitor activity in 11 normal children was 0.94 +/- 0.04 units. During the overt nephrotic syndrome, before initiation of intensive daily corticosteroid treatment, mean values were: prekallikrein, 58.5 +/- 7.24 mumol/ml/hr; and kallikrein inhibitor, 0.35 +/- 0.06 units. After corticosteroid-induced remission occurred, mean values were: plasma prekallikrein, 118.6 +/- 3.2 mumol/ml/hr; and kallikrein inhitor, 0.78 +/- 0.03 mumol/ml/hr. Both parameters were again assayed in 14 of the 21 children after complete cessation of corticosteroid treatment. Plasma prekallikrein was normal, 99.6 +/- 4.8 mumol/ml/hr; but kallikrein inhibitor was still somewhat depressed, 0.84 +/- 0.03 units. A subset of 9 patients had marked depression of plasma prekallikrein to levels less than 20 mumol/ml/hr and essentially undetectable inhibitor activity. Serum alpha-2 macroglobulin was elevated in nephrotic patients: mean value during relapse, 862 +/- 29 mg/100 ml; during corticosteroid-maintaining remission, 615 +/- 29 mg/100 ml. After cessation of corticosteroids, mean serum level was 481 +/- 20 mg/100 ml. The proportional reduction of plasma prekallikrein and kallikrein inhibitor suggested that an enzyme-inhibitor complex formed in vivo, perhaps at a local site of activation in proximity to the glomerular basement membrane. These data suggest that the plasma kinin-generating system may be the host effector mechanism subserving the increased glomerular capillary permeability in the minimal lesion nephrotic syndrome of childhood.