CsA-induced Hypertension Research Articles

Increased Vascular Resistance and Not Salt Retention Characterizes Cyclosporine A–Induced Hypertension: Report in an Anuric Patient

Cyclosporine A (CsA) use is associated with hypertension in most solid-organ transplant recipients. The mechanisms of CsA-induced hypertension have not been fully elucidated and are still controversial. We present a case of CsA-induced hypertension who was anuric and receiving hemodialysis, and in whom noninvasive cardiothoracic bioimpedence revealed elevated systemic vascular resistance without evidence of fluid-volume overload. We briefly discuss the possible mechanisms of CsA-induced hypertension in light of this information.

Lipoic acid supplementation prevents cyclosporine-induced hypertension and nephrotoxicity in spontaneously hypertensive rats

Cyclosporine (CsA) has significantly improved long-term survival after organ transplantations. Hypertension and nephrotoxicity are common side effects during CsA treatment and are aggravated by high salt intake. To examine whether lipoic acid (LA), a natural antioxidant that scavenges reactive oxygen species and regenerates/recycles endogenous antioxidants, could prevent CsA-induced hypertension and nephrotoxicity. Six-week-old spontaneously hypertensive rats (SHR) on a high-sodium diet (NaCl 6%) received CsA [5 mg/kg subcutaneously (s.c.)] alone or in combination with LA (0.5% w/w) for 6 weeks. Blood pressure, arterial functions, and tissue morphology were determined. Immunohistochemistry, quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and high-pressure liquid chromatography were used for kidney and heart samples. CsA induced severe hypertension, cardiac hypertrophy, endothelial dysfunction, and pronounced albuminuria. Histologically, the kidneys showed severe thickening of the media of the afferent arteries with fibrinoid necrosis, perivascular monocyte/macrophage infiltration and nitrotyrosine overexpression. CsA induced the expression of fibrogenic connective tissue growth factor both in the heart and kidneys. The detrimental effects of CsA were associated with upregulation of myocardial atrial natriuretic peptide (ANP) mRNA expression, paradoxical activation of the renin-angiotensin system (RAS), induction of renal reduced nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase, and overexpression of oxidative stress-induced transcription factor NRF2. LA lowered blood pressure, ameliorated cardiac hypertrophy and endothelial dysfunction, and totally normalized albuminuria. In LA-treated rats, renal and cardiac morphologies were indistinguishable from those of SHR controls. CsA-induced myocardial ANP and connective tissue growth factor (CTGF) mRNA overexpression, RAS activation, NADPH oxidase induction, and NRF2 overexpression were prevented by LA. LA induced the mRNA expression of gamma-glutamylcysteine ligase, the rate-limiting enzyme in glutathione synthesis, and markedly increased hepatic cysteine and glutathione concentrations. Our findings suggest a salutary role for lipoic acid supplementation in the prevention of CsA-induced hypertension and nephrotoxicity, and underscore the importance of increased oxidative stress in the pathogenesis of CsA toxicity.

TAURINE ATTENUATES HYPERTENSION AND RENAL DYSFUNCTION INDUCED BY CYCLOSPORINE A IN RATS

Cyclosporine A (CsA) is the first-line immunosuppressant used for the management of solid organ transplantation and autoimmune diseases. Nephrotoxicity is the major limitation of CsA use. Recent evidence suggests that reactive oxygen species (ROS) play an important role in mediating CsA-induced hypertension and nephrotoxicity. Taurine, the major intracellular free beta-amino acid, is known to be an endogenous anti-oxidant and membrane-stabilizing agent. The present study was designed to investigate the effects of taurine on CsA-induced oxidative stress, hypertension and renal dysfunction. 2. Animals were assigned into four groups of seven rats each as follows: (i) control group, receiving vehicle (olive oil; 1 mL/kg, s.c.); (ii) CsA group, given CsA (25 mg/kg per day, s.c.) for 21 days; (iii) taurine group, supplemented with taurine (1% in the drinking water); and (iv) taurine + CsA group, treated with taurine 3 days before and concurrently during CsA injections for 21 days. 3. Cyclosporine A administration elevated blood pressure, reduced serum nitric oxide (NO) levels and deteriorated renal function, as assessed by increased serum creatinine levels and proteinuria and reduced urine flow rate and creatinine clearance compared with vehicle-treated rats. Cyclosporine A induced oxidative stress, as indicated by increased renal tissue concentrations of thiobarbituric acid-reactive substances and reduced concentrations of renal glutathione, glutathione peroxidase and superoxide dismutase. Conversely, no change was noted in renal catalase activity. Moreover, the kidneys of CsA-treated rats showed interstitial inflammation and renal tubular atrophy. 4. Taurine markedly reduced elevated blood pressure, attenuated renal dysfunction and the reduction in serum NO levels and counteracted the deleterious effects of CsA on oxidative stress markers. Furthermore, taurine ameliorated CsA-induced morphological changes. 5. These data clearly indicate the protective potential of taurine against CsA-induced hypertension and nephrotoxicity and suggest a significant contribution of its anti-oxidant property to this beneficial effect.

Effect of preventive and regressive isosorbide 5-mononitrate treatment on catecholamine levels in plasma, platelets, adrenals, left ventricle and aorta in cyclosporin A-induced hypertensive rats

Increased vascular reactivity associated with cyclosporin A (CsA)-induced arterial hypertension might result from increased vasoconstriction and/or decreased vasodilatation. The administration of organic NO donors could have beneficial effects by the NO-cGMP reposition, but there is the risk of sympathetic nervous system worsening by neuro-hormonal counter-regulation. We evaluate the effect of preventive and regressive (curative) isosorbide 5-mononitrate (Is-5-Mn) treatment on blood pressures and on plasma, platelets, adrenals, left ventricle and aorta norepinephrine (NE) and epinephrine (E) contents, assessed by HPLC, in CsA-induced hypertensive rats. Five rat groups were tested: control (orange juice), CsA (5 mg/kg/day) and Is-5-Mn (150 mg/kg/day, bid) groups were treated for 7 weeks; preventive group (Is-5-Mn + CsA): Is-5-Mn during 2 weeks plus 7 weeks of Is-5-Mn + CsA; regressive group (CsA + Is-5-Mn): CsA during 7 weeks plus 5 weeks of CsA + Is-5-Mn. The increased BP in the CsA group was prevented, but was not reverted, by concomitant Is-5-Mn treatment. In the CsA-treated rats, there was a noticeable decrease in left ventricle NE and E contents and aorta NE levels and a moderate increase in circulating catecholamines, without significant effect in the adrenals values. When Is-5-Mn was preventively used, the CsA-induced effect on left ventricle and aorta was prevented. Concomitantly, however, the plasma–platelet catecholamine balance was disrupted, accumulating NE in plasma, whereas E increased in aorta, mimic the single Is-5-Mn-treated group. In opposition, in the group used as regressive Is-5-Mn therapy, the adrenals contents were higher compared with the CsA-group and, simultaneously, the CsA-evoked effects on circulating, left ventricle and aorta catecholamines were not reverted. In conclusion, regressive Is-5-Mn therapy was unable to attenuate CsA-induced catecholamine changes and BP values even worsened. On the contrary, preventive Is-5-Mn treatment prevented the catecholamine changes on left ventricle and aorta, but increased plasma NE and aorta E accumulation. Even though with those effects, hypertension development was totally prevented, suggesting that peripheral SNS per se cannot fully explain CsA-induced hypertension. Furthermore, Is-5-Mn might produce beneficial effects only if preventively employed but, considering the changes on peripheral catecholamine contents, a judicious evaluation of the nitrate therapy impact is recommended in order to avoid further deleterious effects.

Cyclosporine enhances salt sensitivity of body water composition as assessed by impedance among psoriatic patients with normal renal function

Objective Cyclosporine (CsA) therapy may be accompanied by a significant increase in blood pressure, either sodium (Na+) independent or Na+ dependent. The relationship between Na+ intake and body water distribution among patients treated with CsA has not been evaluated. We report the study, by bioelectrical impedance analysis (BIA), of water composition changes after dietary salt manipulations both before and during CsA treatment of psoriatic patients. Methods Ten normotensive psoriatic patients, ages 37 ± 12 years (range, 19 to 54), with normal renal function were included. Each patient was assessed by BIA in 2 phases, before (phase 1) and during (phase 2) CsA therapy (3 mg/kg/day). In both phases, each patient was assessed in basal conditions (basal1 and basal2), on day 7 of a low-sodium diet (LS1 and LS2; 20 mEq/day) and on day 7 of a high-sodium diet (HS1 and HS2; 350 mEq/day). Plasma creatinine (Pcr), urinary volume excretion (Uv), urinary sodium (UNa+), urinary potassium (UK+), urinary osmolality (UOsmo), weight (Wt), resistance (R), reactance (Xc), total body water (TBW), extracellular water (ECW), intracellular water (ICW), Na:K exchangeable (Nae:Ke), phase angle (PA), and body cell mass (BCM) were evaluated. Blood pressure was monitored during 24 hours on the last day of each diet. Paired Student’s t-test was used to analyze the different phases. Results Before CsA treatment, Wt, TBW and Nae:Ke were lower during LS1 than during basal1, whereas TBW was higher during HS1 than during LS1. During CsA, Wt, TBW, ECW, and Nae:Ke were lower during LS2 than during basal2, whereas ICW and PA were higher during LS2 than during basal2. HS2 showed higher TBW, ECW, and Nae:Ke and lower ICW, PA, and BCM than during LS2. Systolic blood pressure was higher during HS2 than during LS2 or HS1. In addition, diastolic blood pressure was higher during HS2 than during HS1. Conclusion Body hydration status was more sensitive to dietary salt fluctuations during CsA treatment than without CsA, and a high-sodium diet seemed to enhance the CsA-induced hypertension side effect. Moreover, patients on low sodium intake under CsA treatment displayed neither any disturbance of body water composition nor any blood pressure change. Our data suggest that a low sodium intake might be very useful in preventing undesirable pressure and volume changes brought about by CsA treatment. Cyclosporine (CsA) therapy may be accompanied by a significant increase in blood pressure, either sodium (Na+) independent or Na+ dependent. The relationship between Na+ intake and body water distribution among patients treated with CsA has not been evaluated. We report the study, by bioelectrical impedance analysis (BIA), of water composition changes after dietary salt manipulations both before and during CsA treatment of psoriatic patients. Ten normotensive psoriatic patients, ages 37 ± 12 years (range, 19 to 54), with normal renal function were included. Each patient was assessed by BIA in 2 phases, before (phase 1) and during (phase 2) CsA therapy (3 mg/kg/day). In both phases, each patient was assessed in basal conditions (basal1 and basal2), on day 7 of a low-sodium diet (LS1 and LS2; 20 mEq/day) and on day 7 of a high-sodium diet (HS1 and HS2; 350 mEq/day). Plasma creatinine (Pcr), urinary volume excretion (Uv), urinary sodium (UNa+), urinary potassium (UK+), urinary osmolality (UOsmo), weight (Wt), resistance (R), reactance (Xc), total body water (TBW), extracellular water (ECW), intracellular water (ICW), Na:K exchangeable (Nae:Ke), phase angle (PA), and body cell mass (BCM) were evaluated. Blood pressure was monitored during 24 hours on the last day of each diet. Paired Student’s t-test was used to analyze the different phases. Before CsA treatment, Wt, TBW and Nae:Ke were lower during LS1 than during basal1, whereas TBW was higher during HS1 than during LS1. During CsA, Wt, TBW, ECW, and Nae:Ke were lower during LS2 than during basal2, whereas ICW and PA were higher during LS2 than during basal2. HS2 showed higher TBW, ECW, and Nae:Ke and lower ICW, PA, and BCM than during LS2. Systolic blood pressure was higher during HS2 than during LS2 or HS1. In addition, diastolic blood pressure was higher during HS2 than during HS1. Body hydration status was more sensitive to dietary salt fluctuations during CsA treatment than without CsA, and a high-sodium diet seemed to enhance the CsA-induced hypertension side effect. Moreover, patients on low sodium intake under CsA treatment displayed neither any disturbance of body water composition nor any blood pressure change. Our data suggest that a low sodium intake might be very useful in preventing undesirable pressure and volume changes brought about by CsA treatment.

Cyclosporine enhances salt sensitivity of body water composition as assessed by impedance among psoriatic patients with normal renal function

Cyclosporine (CsA) therapy may be accompanied by a significant increase in blood pressure, either sodium (Na+) independent or Na+ dependent. The relationship between Na+ intake and body water distribution among patients treated with CsA has not been evaluated. We report the study, by bioelectrical impedance analysis (BIA), of water composition changes after dietary salt manipulations both before and during CsA treatment of psoriatic patients. Ten normotensive psoriatic patients, ages 37 +/- 12 years (range, 19 to 54), with normal renal function were included. Each patient was assessed by BIA in 2 phases, before (phase 1) and during (phase 2) CsA therapy (3 mg/kg/day). In both phases, each patient was assessed in basal conditions (basal1 and basal2), on day 7 of a low-sodium diet (LS1 and LS2; 20 mEq/day) and on day 7 of a high-sodium diet (HS1 and HS2; 350 mEq/day). Plasma creatinine (Pcr), urinary volume excretion (Uv), urinary sodium (UNa+), urinary potassium (UK+), urinary osmolality (UOsmo), weight (Wt), resistance (R), reactance (Xc), total body water (TBW), extracellular water (ECW), intracellular water (ICW), Na:K exchangeable (Nae:Ke), phase angle (PA), and body cell mass (BCM) were evaluated. Blood pressure was monitored during 24 hours on the last day of each diet. Paired Student's t-test was used to analyze the different phases. Before CsA treatment, Wt, TBW and Nae:Ke were lower during LS1 than during basal1, whereas TBW was higher during HS1 than during LS1. During CsA, Wt, TBW, ECW, and Nae:Ke were lower during LS2 than during basal2, whereas ICW and PA were higher during LS2 than during basal2. HS2 showed higher TBW, ECW, and Nae:Ke and lower ICW, PA, and BCM than during LS2. Systolic blood pressure was higher during HS2 than during LS2 or HS1. In addition, diastolic blood pressure was higher during HS2 than during HS1. Body hydration status was more sensitive to dietary salt fluctuations during CsA treatment than without CsA, and a high-sodium diet seemed to enhance the CsA-induced hypertension side effect. Moreover, patients on low sodium intake under CsA treatment displayed neither any disturbance of body water composition nor any blood pressure change. Our data suggest that a low sodium intake might be very useful in preventing undesirable pressure and volume changes brought about by CsA treatment.

Isosorbide-5-mononitrate treatment prevents cyclosporin A-induced platelet hyperactivation and the underlying nitric oxide–cyclic guanosine-3′,5′-monophosphate disturbances

Introduction: The clinical use of cyclosporin A (CsA) is commonly associated with the development of hypertension and increased risk of thromboembolic events. Decreased endothelium-dependent relaxation and increased platelet activation seems to be involved on those side effects, but the underlying mechanisms are not yet elucidated. The present study aimed to evaluate the CsA effect on the platelet NO–cyclic guanosine-3′,5′-monophosphate (cGMP) pathway and the putative benefits of concomitant isosorbide-5-mononitrate (IS-5-MN) administration on CsA-induced hypertension and on platelet hyperactivation. Materials and Methods: Blood pressures, platelet NO synthase activity and cGMP content, intracellular free calcium concentration ([Ca 2+] i) and whole blood platelet aggregation were assessed in three rat groups orally treated, during 7 weeks, with the following diets: orange juice (control group), 5 mg/kg/day of CsA (CsA group) and 150 mg/kg/day, b.i.d., of IS-5-MN for 2 weeks and IS-5-MN plus 5 mg/kg/day of CsA for 7 weeks (IS-5-MN+CsA group). Results: IS-5-MN treatment has prevented hypertension development obtained in the solely CsA-treated rats. CsA treatment has inhibited NOS activity, which was reverted by the concomitant IS-5-MN and CsA administration. On the contrary, platelets from CsA-treated rats had cGMP content increased when compared with the control rats. The variation obtained when ISMN was present was less predominant. Therefore, the organic nitrate treatment has prevented platelet hyperactivation, namely, by decreasing thrombin-evoked [Ca 2+] i and collagen-evoked platelet aggregation, when compared with the solely CsA-treated group. The preventive effect of IS-5-MN was reinforced by electron microscopy studies of platelet activation. Conclusions: By increasing [Ca 2+] i and aggregation, CsA induces platelet hyperactivation and simultaneously increases cGMP content, which might represent a compensatory inhibitory mechanism. The concomitant IS-5-MN treatment prevents the above-mentioned platelet hyperreactivity and tends to normalize the NO–cGMP pathway as well as the development of hypertension.

Role of metoprolol, B1-adrenoceptor antagonist, thromboxane A2 and nitric oxide in CsA-induced hypertension

Chronic treatment with cyclosporine A (CsA), a potent immunosuppressive agent, is associated with the development of arterial hypertension. The effect of CsA on vascular responses was determined in Sprague–Dawley rats and rat aortic rings. Male rats weighing 250–300 g were given either CsA (25 mg/kg/day) in olive oil or vehicle by intraperitoneal (ip) injection for 7 days. CsA administration produced a 42% increase ( P<0.001) in mean arterial pressure (MAP) which reached a plateau after 3 days. The level of both nitrate/nitrite (NO 2/NO 3), metabolites of nitric oxide (NO), decreased by 50% ( P<0.001), but the level of thromboxane A2 (TBXA2) increased by 75% ( P<0.001), in the urine. When 10 −9 M of CsA was added acutely to intact aortic rings from untreated rats, NO 2/NO 3 production decreased by 83% ( P<0.011), but TBXA2 production increased by 86% ( P<0.001). The effects of CsA were reversed both in vivo and in vitro by pretreatment with metoprolol (15 mg/kg/day ip), B1-adrenoceptor antagonist. There were no changes in MAP and tension in rats treated with metoprolol alone. In addition, in aorta of rats that were treated with CsA ip for 7 days, CsA significantly activated protein kinase C (PKC) translocation. This suggests that PKC mediate, in part, CsA-induced hypertension. In summary, CsA inhibits endothelial NO formation, activate PKC, and increase TBXA2 production, with resulting increase in MAP, and this changes can be overcome by pretreatment with metoprolol.

HMG-CoA reductase inhibition and PPAR- alpha activation both inhibit cyclosporin A induced endothelin-1 secretion in cultured endothelial cells.

The use of cyclosporin A (CsA) in solid organ transplantation has been shown to be associated with the development of hypertension and nephrotoxicity. Several mechanisms, including endothelin (ET)-1-mediated systemic vasoconstriction, are considered to be responsible for CsA-induced hypertension. This study shows that: (i) incubation of CsA (1 microg) with bovine aortic endothelial cells leads to increased ET secretion by+40%; (ii) the use of compactin, the 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor and fibric acid, the peroxisome-proliferator-activated receptor (PPAR)-alpha activator, inhibit the CsA-induced ET secretion to the level below the basal ET secretion, by -32% and -26% respectively; (iii) both inhibitions were reversed by the addition of mevalonate, suggesting communication between the HMG-CoA reductase product and PPAR-alpha pathway. The present findings may be of significant clinical relevance, since statins and fibrates beyond their hypolipidaemic action may represent a potential therapeutic tool in the treatment or prophylaxis of CsA-associated side effects. Furthermore, we suggest that the mevalonate metabolism would interfere with PPAR-alpha activity.

Interaction of cyclosporine A and the renin-angiotensin system; new perspectives.

Despite extensive research, the exact mechanisms of cyclosporine A (CsA)-induced hypertension and nephrotoxicity remain obscure. Several lines of evidence suggest an involvement of the renin-angiotensin system (RAS) in CsA toxicity, but the issue is still controversial in more ways than one. Some interesting data of the interaction of CsA and RAS have been presented by us and others during the last years. In rats, activation of RAS by CsA is a consistent finding while the results from clinical studies show controversial results. The mechanisms of activation of RAS may be multifactorial. CsA increases renin release directly from juxtaglomerular cells. However, RAS activation may at least partly account for glomerular ischemia by vasoconstriction. A totally different view about the interaction of CsA and RAS has recently been presented. CsA antagonised the harmful effects of RAS over-expression on renal damage in double transgenic rats harbouring human renin and angiotensinogen genes. The protection was due to anti-inflammatory properties of CsA by inhibition of interleukin-6 and inducible nitric oxide synthase (iNOS) expression. Other studies have confirmed the inhibitory effect of CsA on iNOS. Calcium antagonists have been proposed to be the antihypertensive drugs of choice in treatment of CsA-induced hypertension because of their favourable haemodynamic effects on the kidneys. However, because angiotensin II plays a major role in the development of CsA-induced structural renal damage, pharmacological inhibition of RAS in CsA-treatment may have some beneficial effects beyond blood pressure control.

Nitric oxide in CsA-induced hypertension: role of β -adrenoceptor antagonist and thromboxane A2

Cyclosporine A (CsA) is an immunosuppressive agent, which also causes hypertension. The effect of CsA on vascular responses was determined in Sprague-–Dawley rats and rat aortic rings. Male rats weighing 250-–300 g were given either CsA (25 mg/kg/day) in olive oil or vehicle by intraperitoneal (ip) injection for 7 days. CsA administration produced a 42% increase (P<0.001) in mean arterial pressure (MAP) which reached a plateau after 3 days. The level of both nitrate/nitrite (NO2/NO3), metabolites of nitric oxide (NO), decreased by 50% (P<0.001), but the level of thromboxane A2-(TBXA2) increased by 75% (P<0.001), in the urine. When 10−9M of CsA was added acutely to intact aortic rings from untreated rats, NO2/NO3 production decreased by 83% (P<0.011), but TBXA2- production increased by 86% (P<0.001). The effects of CsA were reversed both in vivo and in vitro by pretreatment with propranolol (15 mg/kg/day ip), β -adrenoceptor antagonist. There were no changes in MAP and tension in rats treated with prop alone. In addition, in aorta of rats that were treated with CsA ip for 7 days, CsA significantly activated protein kinase C (PKC) translocation. This suggests that PKC mediate, in part, CsA-induced hypertension. In summary, CsA inhibits endothelial NO formation, activate PKC, and increase TBXA2 production, with resulting increase in MAP, and this changes can be overcome by pretreatment with propranolol.

The endothelin receptor antagonist, L-754,142 does not prevent cyclosporine A-induced osteopenia in rats.

Cyclosporine A (CsA) is a potent immunosuppressive agent widely used to prevent allograft rejection. In vivo administration of CsA is associated with the development of high-turnover osteopenia. Endothelin-1 (ET), a vasoconstrictive peptide, has been implicated in CsA-induced nephrotoxicity and hypertension. Recent evidence suggests that endothelin plays a pivotal role in bone metabolism. The present study was designed to investigate whether L-754,142 (ETRA), the combined endothelin A and B receptor antagonist, when given to rats, would favorably modify the bone loss caused by CsA. Fifty, 5-month-old male Sprague-Dawley rats were randomly divided into five groups of 10 rats each. The first group served as a basal control. The remaining four groups received, by daily gavage for 28 days, (1) a combined CsA and ETRA vehicle, (2) CsA, 10 mg/kg, (3) ETRA, 30 mg/kg, and (4) CsA, 10 mg/kg and ETRA, 30 mg/kg, respectively. Rats were weighed and venous blood was collected on days 0, 14, 28 for determination of BUN, creatinine, calcium, PTH, osteocalcin, and 1,25(OH)2 D. Tibiae, after double labeling, were removed following sacrifice for histomorphometry. Both CsA-treated rats and CsA/ETRA-treated rats demonstrated trabecular osteopenia with raised serum osteocalcin, and 1,25(OH)2D levels when compared to control animals (P < 0.05). Rats given CsA alone developed renal impairment, as shown by an increased BUN. The combination group did not develop renal impairment. The results suggest that endothelin may contribute to the development of CsA-induced nephrotoxicity, which was prevented by ETRA, but does not seem to play a role in CsA-induced osteopenia.

Dietary potassium and magnesium supplementation in cyclosporine-induced hypertension and nephrotoxicity

Cyclosporine A (CsA)-induced hypertension and nephrotoxicity are aggravated by high sodium intake. Accumulating evidence suggests that potassium and magnesium supplementation could protect against the detrimental effects of dietary salt. In the present study, we tested the hypothesis of whether concurrent supplementation with potassium and magnesium could protect against the development of CsA-induced hypertension and nephrotoxicity more effectively than supplementation with one mineral alone. Eight-week-old spontaneously hypertensive rats (SHRs) were divided into four groups (N = 10 in each group): (1) CsA group (5 mg/kg subcutaneously) receiving high-sodium diet (Na 2.6%, K 0.8%, Mg 0.2% wt/wt); (2) CsA group receiving a high-sodium, high-potassium diet (Na 2.6%, K 2.4%, Mg 0.2%); (3) CsA group receiving high-sodium, high-magnesium diet (Na 2.6%, K 0.8%, Mg 0.6%); and (4) CsA group receiving high-sodium, high-potassium, high-magnesium diet (Na 2.6%, K 2.4%, Mg 0.6%). CsA induced severe hypertension and deteriorated renal functions in SHRs on high-sodium diet. Histologically, the kidneys showed severe thickening of the media of the afferent artery with fibrinoid necrosis. Potassium supplementation lowered blood pressure (198 +/- 5 vs. 212 +/- 2 mm Hg, P < 0.05) and partially prevented the development of proteinuria (-25%, P < 0.05). Magnesium supplementation decreased blood pressure to the same extent but improved renal functions more effectively than potassium. The greatest protection against CsA toxicity was achieved when dietary potassium and magnesium supplementations were combined. Urinary N-acetyl-beta-D-glucosaminidase (NAG) excretion, a marker for renal proximal tubular damage, increased progressively in CsA-treated SHRs on the high-sodium diet. Neither potassium nor magnesium influenced urinary NAG excretion. We also estimated the activity of the renal dopaminergic system by measuring 24-hour urinary dopamine excretion rates. CsA suppressed the renal dopaminergic system during high-sodium diet. Magnesium supplementation, alone and in combination with potassium, protected against the development of renal dopaminergic deficiency in CsA-treated SHRs on high-sodium diet. Magnesium supplementation increased plasma-free ionized magnesium (iMg) and bone magnesium by 50 and 16%, respectively. Our findings indicate that both potassium and magnesium supplementations showed beneficial effects against CsA-induced hypertension and nephrotoxicity. The protective effect of magnesium clearly exceeded that of potassium. The greatest protection against CsA toxicity was achieved when potassium and magnesium were combined. We also provide evidence that the development of CsA-induced glomerular, tubular, and vascular lesions are associated with renal dopaminergic deficiency.

Calcineurin inhibitors cause renal afferent activation in rats: a novel mechanism of cyclosporine-induced hypertension

Inhibition of calcineurin-mediated signaling in T lymphocytes is a major mechanism of cyclosporine A (CsA)-induced immunosuppression, and previous rat studies have suggested that inhibition of calcineurin-mediated signaling in central neuronal pools involved in blood pressure regulation plays an important role in causing acute CsA-induced hypertension. However, a central neural mechanism is difficult to reconcile with other data suggesting that CsA-induced hypertension is due to activation of renal and other subdiaphragmtic visceral afferents that reflexively increase efferent sympathetic nerve activity. Accordingly, we now have revised our hypothesis to suggest that CsA stimulates renal afferents by a calcineurin-dependent process. To test this new hypothesis, in anesthetized rats we recorded arterial pressure and multifiber afferent renal nerve activity from the cut distal end of the renal nerve before, during, and after intravenous infusion of either CsA (5 mg/kg over 20 min, n = 8), FK506 (0.15 mg/kg, n = 7), another potent calcineurin inhibitor that is structurally unrelated to CsA, or rapamycin (0.15 mg/kg, n = 4), a structural analog of FK506 that has no effect on calcineurin. We found that renal afferent discharge was increased markedly by intravenous FK506, as well as CsA, but unaffected by rapamycin (or vehicle), indicating calcineurin mediation. After infusion of either calcineurin inhibitor, afferent renal nerve activity remained elevated for up to 2 h, paralleling the prolonged increase in blood pressure. Thus, the major new conclusion of this study is that, in contrast to what has been assumed previously, calcineurin inhibitors enhance sympathetic neurotransmission by a novel action localized to visceral sensory nerve endings rather than to nerve cell bodies or central synapses. In the rat, calcineurin-dependent activation of renal afferents appears to be the primary mechanism producing the large blood-pressure–raising effect of CsA. Because the data suggest that the major side-effect of CsA and FK506—hypertension—is inexorably linked to calcineurin inhibition in extralymphoid tissue, development of agents that selectively inhibit calcineurin only in T lymphocytes could eliminate this important secondary form of hypertension.

Cyclosporin-induced endothelial dysfunction and hypertension: are nitric oxide system abnormality and oxidative stress involved?

Abstract Hypertension is a major side effect of cyclosporin (CsA). While the mechanism(s) responsible are unclear, CsA-induced endothelial dysfunction and CsA-induced hypertension have been attributed to the CsA effect on the endothelial-derived factors controlling vasomotor tone. Endothelial nitric oxide (NO) is crucial in the maintenance of a state of basal vasodilation, and recent studies have suggested an NO-mediated counterregulatory mechanism protective from CsA-induced vasoconstriction. Our study evaluates endothelial nitric oxide synthase (ecNOS) gene status (PCR analysis) and plasma levels of NO metabolites (ELISA) in kidney and heart transplant patients under chronic CsA treatment with CsA-induced hypertension. Since CsA increases superoxide production, which metabolises NO, plasma hydroperoxides from cholesterol esters and from triglycerides and peroxynitrite were also evaluated (HPLC) as an index of the presence of superoxides and of “oxidative stress”. Quantification of monocyte ecNOS mRNA and NO metabolites plasma levels from patients and controls (C) demonstrated NO system upregulation in patients notwithstanding the hypertension. The mean ecNOS to β-actin ratio was 1.80 ± 0.85 in patients vs 0.40 ± 0.09 in C (P < 0.04). NO metabolites were 34.03 ± 14.32 μM in patients vs 11.53 ± 5.64 μM in C (P < 0.001). Hydroperoxides from cholesterol esters and from triglycerides were also increased in patients, 3.4 ± 1.4 vs 1.3 ± 0.6 integrated area units (i. a. u.), P < 0.007 and 10.6 ± 6.4 vs 1.3 ± 0.8 i.a.u., P < 0.008, respectively, as well as the peroxynitrite plasma level, 0.32 ± 0.11 μM/l vs undetectable in C. This study confirms a CsA-induced NO system upregulation in transplanted patients. However, the NO-mediated counterregulatory system to CsA-induced vasoconstriction, present in normals, could be canceled in patients by CsA-induced superoxide (O2−) and free radical production which, by increasing NO metabolism, could contribute to CsA-induced vasoconstriction and hypertension and predispose to atherosclerosis.

Cyclosporin-induced endothelial dysfunction and hypertension: are nitric oxide system abnormality and oxidative stress involved?

Hypertension is a major side effect of cyclosporin (CsA). While the mechanism(s) responsible are unclear, CsA-induced endothelial dysfunction and CsA-induced hypertension have been attributed to the CsA effect on the endothelial-derived factors controlling vasomotor tone. Endothelial nitric oxide (NO) is crucial in the maintenance of a state of basal vasodilation, and recent studies have suggested an NO-mediated counterregulatory mechanism protective from CsA-induced vasoconstriction. Our study evaluates endothelial nitric oxide synthase (ecNOS) gene status (PCR analysis) and plasma levels of NO metabolites (ELISA) in kidney and heart transplant patients under chronic CsA treatment with CsA-induced hypertension. Since CsA increases superoxide production, which metabolises NO, plasma hydroperoxides from cholesterol esters and from triglycerides and peroxynitrite were also evaluated (HPLC) as an index of the presence of superoxides and of "oxidative stress". Quantification of monocyte ecNOS mRNA and NO metabolites plasma levels from patients and controls (C) demonstrated NO system upregulation in patients notwithstanding the hypertension. The mean ecNOS to beta-actin ratio was 1.80 +/- 0.85 in patients vs 0.40 +/- 0.09 in C (P < 0.04). NO metabolites were 34.03 +/- 14.32 microM in patients vs 11.53 +/- 5.64 microM in C (P < 0.001). Hydroperoxides from cholesterol esters and from triglycerides were also increased in patients, 3.4 +/- 1.4 vs 1.3 +/- 0.6 integrated area units (i. a. u.), P < 0.007 and 10.6 +/- 6.4 vs 1.3 +/- 0.8 i. a. u., P < 0.008, respectively, as well as the peroxynitrite plasma level, 0.32 +/- 0.11 microM/l vs undetectable in C. This study confirms a CsA-induced NO system upregulation in transplanted patients. However, the NO-mediated counterregulatory system to CsA-induced vasoconstriction, present in normals, could be canceled in patients by CsA-induced superoxide (O2-) and free radical production which, by increasing NO metabolism, could contribute to CsA-induced vasoconstriction and hypertension and predispose to atherosclerosis.

Nitric oxide in cyclosporine A-induced hypertension: role of protein kinase C

Chronic treatment with cyclosporine A (CsA), an immunosuppressive agent, causes hypertension. The effect of CsA on vascular responses was determined in Sprague-Dawley rats and isolated rat aortic rings. Male rats weighing 250 to 300 g were given either CsA (25 mg/kg/day) in olive oil or vehicle by intraperitoneal injection for 7 days. Cyclosporine A administration produced a 42% increase ( P < .001) in mean arterial pressure (MAP), which reached a plateau after 3 days. Conversely, the level of both nitrate/nitrite (NO 2/NO 3), metabolites of nitric oxide (NO), and 3′, 5′ cyclic guanosine monophosphate (cGMP), which mediates NO action, decreased by 50% ( P < .001) and 35% ( P < .001), respectively, in the urine. Thoracic aortic rings from rats treated with CsA, and precontracted with endothelin (10 −9 mol/L), showed a 35% increase ( P < .001) in tension, whereas acetylcholine-induced (Ach; 10 −9 mol/L) endothelium-dependent relaxation was inhibited 65% ( P < .001) compared with untreated rats. This response was similar to that of aortic rings, denuded of endothelium, from untreated rats in which Ach-induced relaxation was completely abolished ( P < .001). Ach-induced formation of both NO 2/NO 3 and cGMP by both denuded and CsA-treated aortic rings was inhibited 95% ( P < .001) and 65% P < .001), respectively, compared with intact aortic rings. The effects of CsA were reversed both in vivo and in vitro by pretreatment with l-arginine (L-Arg; 10 mg/kg/day intraperitoneally), the precursor of NO. There were no changes in MAP and tension in rats treated with L-Arg alone. In addition, in the aorta of rats that were treated intraperitoneally with CsA for 7 days, CsA significantly activated protein kinase C (PKC) translocation and decreased NO 2/NO 3 production. This suggest that PKC mediates, in part, CsA-induced hypertension. In summary, CsA activates PKC, which inhibits endothelial NO formation, with resulting increases in MAP and tension, and this inhibition can be overcome by L-Arg administration.

Cyclosporine A up-regulates angiotensin II receptors and calcium responses in human vascular smooth muscle cells

The most widely used immunosuppressive drug for preventing graft rejection and treating autoimmune diseases is currently cyclosporine A (CsA). However, CsA also causes vasoconstriction, which is considered to be at the origin of CsA-induced nephrotoxicity and hypertension. To evaluate the cellular basis for these side effects, we studied the influence of CsA on the regulation of the free cytosolic Ca2+ concentration ([Ca2+]c) in cultured human vascular smooth muscle cells (SMCs). SMCs were isolated from the medial layer of human aorta. [Ca2+]c regulation was studied by fluorimetry with fura 2 and by measuring 45Ca2+ effluxes. Angiotensin II (Ang II) receptors were detected by [125I]Ang II binding. Pretreatment of human SMCs for 24 hours with CsA in its therapeutic concentration range (0. 1 to 10.0 microM) had no effect on basal [Ca2+]c, but increased the [Ca2+]c elevation and 45Ca2+ efflux when cells were stimulated with Ang II. Half-maximal effects occurred at approximately 1 microM CsA. The CsA effects on [Ca2+]c were accompanied by a nearly twofold increase in Ang II receptor number, whereas no change in affinity to Ang II was observed. CsA did not alter endothelin-1- or thapsigargin-induced 45Ca2+ efflux. Increases in both Ca2+ responses and [125I]Ang II binding were attenuated by the transcriptional inhibitor actinomycin D. The effects of CsA did not appear to be mediated by calcineurin inhibition because cyclosporine H, which is not immunosuppressive, also increased the Ang II-induced 45Ca2+ efflux. These data suggest that CsA preferentially up-regulates the transcription of Ang II receptors, which very likely leads to vasoconstriction in vivo and could be at the origin of CsA-induced hypertension and nephrotoxicity in humans.

Role of Nitric Oxide and Reactive Oxygen Species in Cyclosporin-Induced Hypertension in Transplanted Patients.

637 Hypertension is a major side effect of Cyclosporin (CsA). While mechanism/s responsible are unclear, CsA-induced endothelial dysfunction and CsA-induced hypertension has been attributed to CsA effect on the endothelial-derived factors controlling vasomotor tone. Endothelial NO is crucial in the manteinance of a state of basal vasodilation and recently an NO mediated counterregulatory mechanism protective from CsA-induced vasoconstriction has been suggested. Our study evaluates ecNOS gene status (Polymerase Chain Reaction analysis) and plasma level of NO metabolites (NO2−/NO3−) (ELISA) in 10 heart (HT) and 10 kidney (KT) transplanted patients under chronic CsA treatment with CsA-induced hypertension. Diagnosis of CsA-induced hypertension in HT was based on the development of hypertension only after CsA administration and without improvement after the tapering of steroids associated to CsA. In KT, by the development of hypertension only after transplant, for those normotensive before the transplant, and by the worsening of a preexisting hypertension notwithstanding the tapering of steroids associated to CsA treatment. Since CsA increases superoxide anions, which combine with NO to yield peroxynitrite, a powerful oxidant, plasma peroxynitrites and hydroperoxides were evaluated (HPLC) as index of the presence of superoxides and free radicals and therefore of oxidative stress. 10 healthy subjects were used as controls (C). Quantification of monocyte ecNOS mRNA from patients and controls demonstrated NO system up regulation in patients notwithstanding the hypertension. The mean ecNOS to β-actin ratio was 1.80±0.85 in patients vs 0.40±0.09 in C (p<0.04). (NO2−/NO3−) plasma level was 34.03±14.32 mM in patients vs 11.53±5.64, p<0.001. Hydroperoxides were also increased in patients vs C: 3.4±1.4 i.a.u. vs 1.3±0.6, p<0.007 (from colesterol esters) and 10.6±6.4 vs 1.3±0.8, p<0.008 (from triglycerides) as well as peroxynitrite plasma level (0.32±0.11 mM/L vs undetectable in C. This study confirms a CsA induced NO system up regulation in transplanted patients. However, the counterregolatory system to CsA-induced vasoconstriction could be canceled in patients by CsA induced superoxides and free radicals production which, increasing NO metabolism and therefore reducing its efficacy as vasorelaxing factor could contribute to CsA induced vasoconstriction and hypertension and promoting remodelling processes, predispose to long term hypertensive complications such as cardiac hypertrophy and atherosclerosis.

Effects of ACE Inhibition on Cyclosporine A-Induced Hypertension and Nephrotoxicity in Spontaneously Hypertensive Rats on a High-sodium Diet

Cyclosporine A (CsA)-induced hypertension has been shown to be dependent on the level of dietary salt. The present study assessed the role of the renin-angiotensin system in the development of CsA-induced hypertension and nephrotoxicity in spontaneously hypertensive rats (SHR) on a high-sodium diet. In addition, we examined whether ACE inhibition prevents the detrimental effects of CsA on blood pressure, kidney function and vascular morphology in SHR on high sodium intake. Eight-week-old SHR were divided into three different groups (n