marinobufagenin and Disease-Models--Animal

Pre-eclampsia (preE), a pregnancy disorder with the de novo onset of hypertension and proteinuria after 20 weeks of gestation, has multiple triggers that initiate pathophysiologic mechanisms. This review addresses translational aspects of preE by synthesizing information on preE pathogenesis, describing diagnostic biomarkers that predict disease, and suggesting strategies to lessen adverse outcomes. Key to this understanding is the role of cardiotonic bufodienolides, with marinobufagenin (MBG) as the prototype, and angiogenic factors in preE pathogenesis. Data from a rat model believed to mimic human preE show that urinary excretion of MBG increases before the onset of hypertension and proteinuria and that affected animals have an increased vascular leakage and blood brain barrier permeability. Angiogenic imbalance occurs with the onset of the syndrome in this model. Also, we report that MBG levels in preE patients exceed those in normal pregnancy and that angiogenic factors are altered in patients showing signs and symptoms of overt disease. In vitro administration of MBG inhibits cytotrophoblast function and triggers hyperpermeability in endothelial cell monolayers. We advance the hypotheses that MBG precedes preE; MBG causes disruption of tight junction proteins leading to vascular leak via activation of MAPK which triggers apoptotic mechanisms resulting in further endothelial dysfunction leading to edema with the release of angiogenic factors. This review provides new evidence about the role of MBG and vasoactive intermediates in preE pathogenesis including the neurologic sequela and may reveal new therapeutic targets for the prevention of preE complications.

Topics: Animals; Biomarkers; Bufanolides; Disease Models, Animal; Female; Humans; Neovascularization, Pathologic; Pre-Eclampsia; Pregnancy; Rats; Translational Research, Biomedical; Vasoconstrictor Agents

The purpose of this review is to provide information detailing the existing evidence with regard to the hypothesis that marinobufagenin (MBG) is an important etiologic and predictive factor in preeclampsia (PE). In addition, evidence describing the role of the antagonist to MBG, resibufogenin (RBG), in the prevention and/or treatment of this disorder is provided.. The studies outlined were performed in an animal model of PE, in in vitro experiments, and in human studies.. Data have been obtained that strongly support the hypothesis that ~60 to 70% of PE patients demonstrate elevations in urinary and serum MBG levels. In the animal model, the entire syndrome can be prevented by the administration of RBG beginning early in pregnancy.. Expanded human trials of MBG as a predictor of the later development of PE are warranted as are studies of the efficacy and safety of RBG as a preventative/therapy.

Topics: Animals; Blood Pressure; Bufanolides; Capillary Permeability; Cardenolides; Clinical Trials as Topic; Disease Models, Animal; Endothelium, Vascular; Female; Hematocrit; Humans; Pre-Eclampsia; Pregnancy; Saponins; Treatment Outcome

Previous studies implicated cardiotonic steroids, including Na/K-ATPase inhibitor marinobufagenin (MBG), in the pathogenesis of preeclampsia (PE). We demonstrated that MBG induces fibrosis via mechanism involving inhibition of Fli1, a nuclear transcription factor and a negative regulator of collagen-1 synthesis. We hypothesized that PE blockade of increased MBG with antibody would lessen the fibrosis of umbilical arteries and lower the blood pressure in rats with PE.. We tested 36 pregnant Sprague-Dawley rats in which 12 were made hypertensive by 1.8% Na supplementation (days 6-19 of gestation), 12 pregnant rats served controls. At day 19, PE rats received one intraperitoneal injection of polyclonal anti-MBG-4 antibody (0.5 ug/ml) for 4 hours.. PE was associated with higher blood pressure (117 ± 2 vs. 107 ± 2 mm Hg; P < 0.01), plasma MBG levels (1.54 ± 0.34 vs. 0.49 ± 0.11 nmol/L; P < 0.01), protein excretion (26 vs. 12 mg/24 hours), sFlt-1 (3-fold), decrease in Fli1 (7-fold) and increase in collagen-1 in aorta (4-fold) vs. control rats (all P < 0.01). In 12 rats treated with polyclonal anti-MBG-4 antibody blood pressure dropped (93 ± 3 mm Hg) and Fli1 was decreased much less (2-fold; P < 0.01 vs. nontreated rats).. These results demonstrate that in experimental PE elevated MBG level is implicated in umbilical fibrosis via suppression of Fli1.

Topics: Animals; Antibodies; Antihypertensive Agents; Blood Pressure; Bufanolides; Disease Models, Animal; Female; Fibrosis; Pre-Eclampsia; Pregnancy; Proto-Oncogene Protein c-fli-1; Rats, Sprague-Dawley; Sodium Chloride, Dietary; Sodium-Potassium-Exchanging ATPase; Umbilical Arteries; Up-Regulation

Background Elevated levels of an endogenous Na/K-ATPase inhibitor marinobufagenin accompany salt-sensitive hypertension and are implicated in cardiac fibrosis. Immunoneutralization of marinobufagenin reduces blood pressure in Dahl salt-sensitive (Dahl-S) rats. The effect of the anti-marinobufagenin monoclonal antibody on blood pressure, left ventricular (LV) and renal remodeling, and gene expression were investigated in hypertensive Dahl-S rats. Methods and Results Dahl-S rats were fed high NaCl (8%, HS; n=14) or low NaCl (0.1%, LS; n=14) diets for 8 weeks. Animals were administered control antibody (LS control antibody, LSC; HS control antibody, HSC; n=7 per group) or anti-marinobufagenin antibody once on week 7 of diet intervention (n=7 per group). Levels of marinobufagenin, LV, and kidney mRNAs and proteins implicated in profibrotic signaling were assessed. Systolic blood pressure was elevated (211±8 versus 133±3 mm Hg,

Topics: Animals; Blood Pressure; Blotting, Western; Bufanolides; Disease Models, Animal; Echocardiography; Enzyme Inhibitors; Gene Expression Regulation; Heart Ventricles; Hypertension; Male; Rats; Rats, Inbred Dahl; RNA; Transforming Growth Factor beta; Ventricular Remodeling

We have investigated serum levels of immunoreactive marinobufagenin (MBG) in 16- to 20-week-old spontaneously hypertensive rats (SHRs)-A3 and in the normotensive Wistar-Kyoto (WKY) rat strain in the absence of salt loading, and we have investigated the genetic control of serum MBG.. We genotyped the F2 progeny of an SHR-A3×WKY intercross using a genome-wide panel of 253 single-nucleotide polymorphism markers that were dimorphic between SHR-A3 and WKY and measured serum MBG by ELISA. Serum MBG levels were lower in SHR-A3 than WKY rats (0.39±0.07 and 1.27±0.40 nmol/L, respectively), suggesting that MBG may not play a role in the markedly divergent blood pressure measured by telemetry in rats of these 2 strains (SHR-A3 and WKY, 198.3±4.43 and 116.8±1.51 mm Hg, respectively). The strain difference in serum MBG was investigated to determine whether genomic regions influencing MBG might be identified by genetic mapping. Quantitative trait locus mapping indicated a single locus influencing serum MBG in the region of chromosome 6q12. Homozygosity of WKY alleles at this locus was associated with increased serum MBG levels. We surveyed whole genome sequences from our SHR-A3 and WKY lines, seeking coding sequence variation between SHR-A3 and WKY within the mapped locus that might explain the inherited strain difference in serum MBG.. We identified amino acid substitution in the sterol transport protein Abcg5, present in SHR-A3, but absent in WKY, that is a potential mechanism influencing MBG levels.

Topics: Animals; ATP Binding Cassette Transporter, Subfamily G, Member 5; Biomarkers; Blood Pressure; Bufanolides; Crosses, Genetic; Disease Models, Animal; Genetic Predisposition to Disease; Genome-Wide Association Study; Homozygote; Hypertension; Lipoproteins; Male; Phenotype; Polymorphism, Single Nucleotide; Quantitative Trait Loci; Rats, Inbred SHR; Rats, Inbred WKY

The acute respiratory distress syndrome (ARDS) represents a major challenge for clinicians as well as basic scientists. The mortality rate for ARDS has been maintained within the range of 40-52%. The authors have examined the involvement of the "cardiotonic steroids" in the pathogenesis and therapy of ARDS. We have studied the possible role of the bufadienolide, marinobufagenin (MBG), in the pathogenesis of ARDS in both a rat model of ARDS and in patients afflicted with that disorder. In addition, the potential therapeutic benefit of an antagonist of MBG, resibufogenin (RBG), in an animal model has been evaluated.. A syndrome resembling human ARDS was produced in the rat by exposing the animals to 100% oxygen for 48 h. In other animals, RBG was administered to these "hyperoxic" rats, and the serum MBG was measured. In human ICU patients, urinary samples were examined for levels of MBG, and the values were compared to those obtained from other ICU patients admitted with diagnoses other than ARDS.. (1) Exposure of rats to hyperoxia produced a histologic picture which resembled that of human ARDS. (2) Serum levels of MBG in the "hyperoxic" rats substantially exceeded those obtained in animals exposed to ambient oxygen levels and were reduced to normal by RBG. (3) In ARDS patients, substantial elevations in urinary MBG were obtained compared to those in non-ARDS ICU patients.. MBG may serve as an important biomarker for the development of ARDS, and RBG may represent a preventative/therapy in this disorder.

Topics: Acute Lung Injury; Animals; Biomarkers; Bufanolides; Case-Control Studies; Disease Models, Animal; Humans; Hyperoxia; Pulmonary Alveoli; Pulmonary Edema; Rats; Respiratory Distress Syndrome; Up-Regulation

Marinobufagenin (MBG) is an endogenous Na/K-ATPase inhibitor, a natriuretic and a vasoconstrictor. MBG is implicated in salt-sensitive hypertension, cardiac hypertrophy, and initiate the pro-fibrotic signaling. Previously it was demonstrated that immunoneutralization of an endogenous MBG by 3E9 anti-MBG-antibody (mAb) in vivo lowered blood pressure (BP) and reversed cardiac fibrosis in salt-sensitive, and in partially nephrectomized rats. In the present study, we investigated whether mAb alleviates vascular remodeling induced in normotensive rats on high salt intake.. Wistar rats (5 months old) received normal (CTRL; n = 8) or high salt intake (2% NaCl in drinking water) for 4 weeks ( n = 16). Rats from the group on a high salt intake were administered vehicle (SALT; n = 8) or mAb (50 µg/kg) (SALT-AB; n = 8) during the last week of high salt diet. BP, erythrocyte Na/K-ATPase activity, levels of MBG in plasma and 24-hour urine, and sensitivity of aortic explants to the vasorelaxant effect of sodium nitroprusside (SNP) were measured. Aortic collagen abundance was determined immunohistochemically.. In SALT vs. CTRL, heightened levels of MBG were associated with inhibition of erythrocyte Na/K-ATPase in the absence of BP changes. High salt intake was accompanied by a 2.5-fold increase in aortic collagen abundance and by a reduction of sensitivity of aortic explants to the vasorelaxant effect of SNP following endothelin-1-induced constriction. In the SALT-AB group, all NaCl-mediated effects were reversed by immunoneutralization of MBG.. High salt intake in young normotensive rats can induce vascular fibrosis via pressure-independent/MBG-dependent mechanisms, and this remodeling is reduced by immunoneutralization of MBG.

Topics: Animals; Antibodies, Monoclonal; Aorta; Aortic Diseases; Bufanolides; Collagen; Disease Models, Animal; Fibrosis; Male; Rats, Wistar; Sodium, Dietary; Vasodilation; Vasodilator Agents

Previous studies from this laboratory have demonstrated that the bufodienolide, marinobufagenin, causes a syndrome in the pregnant rat that resembles human preeclampsia. Furthermore, marinobufagenin urinary excretion is elevated in approximately 85% of preeclamptic patients. Resibufagenin, an antagonist to marinobufagenin, completely prevents the syndrome (hypertension, proteinuria, and intrauterine growth restriction) if given from early pregnancy.. We investigated the effects of another bufodienolide, cinobufatalin, to determine if it, likewise, could induce the rat "preeclamptic" syndrome, which it did. We then examined whether resibufagenin could prevent the syndrome due to cinobufatalin.. Resibufagenin improved hypertension but not proteinuria, and did not prevent uterine growth restriction.. We conclude that more than one bufodienolide may induce the preeclamptic syndrome and that each may require a specific antagonist to prevent (or treat) the syndrome.

Topics: Animals; Bufanolides; Disease Models, Animal; Female; Fetal Growth Retardation; Hypertension; Pre-Eclampsia; Pregnancy; Proteinuria; Rats

Plasma levels of cardiotonic steroids are elevated in volume-expanded states, such as chronic kidney disease, but the role of these natriuretic hormones in subjects with heart failure (HF) is unclear. We sought to determine the prognostic role of the cardiotonic steroids marinobufagenin (MBG) in HF, particularly in relation to long-term outcomes.. We first measured plasma MBG levels and performed comprehensive clinical, laboratory, and echocardiographic assessment in 245 patients with HF. All-cause mortality, cardiac transplantation, and HF hospitalization were tracked for 5 years. In our study cohort, median (interquartile range) MBG was 583 (383-812) pM. Higher MBG was associated with higher myeloperoxidase (r=0.42, P<0.0001), B-type natriuretic peptide (r=0.25, P=0.001), and asymmetrical dimethylarginine (r=0.32, P<0.001). Elevated levels of MBG were associated with measures of worse right ventricular function (RV s', r=-0.39, P<0.0001) and predicted increased risk of adverse clinical outcomes (MBG≥574 pmol/L: hazard ratio 1.58 [1.10-2.31], P=0.014) even after adjustment for age, sex, diabetes mellitus, and ischemic pathogenesis. In mice, a left anterior descending coronary artery ligation model of HF lead to increases in MBG, whereas infusion of MBG into mice for 4 weeks lead to significant increases in myeloperoxidase, asymmetrical dimethylarginine, and cardiac fibrosis.. In the setting of HF, elevated plasma levels of MBG are associated with right ventricular dysfunction and predict worse long-term clinical outcomes in multivariable models adjusting for established clinical and biochemical risk factors. Infusion of MBG seems to directly contribute to increased nitrative stress and cardiac fibrosis.

Topics: Adult; Aged; Animals; Biomarkers; Bufanolides; Cohort Studies; Disease Models, Animal; Female; Heart Failure; Heart Transplantation; Hospitalization; Humans; Male; Mice; Mice, Inbred C57BL; Middle Aged; Outcome Assessment, Health Care; Predictive Value of Tests; Stroke Volume; Survival Analysis; Ventricular Dysfunction, Right

Marinobufagenin (MBG) is a cardiotonic steroid that is increased in preeclampsia. An analog of MBG, resibufogenin (RBG), prevents the development of preeclampsia in a rat model. Oxidative stress is a concomitant of endothelial dysfunction in the latter disorder. The objective of the current studies was to evaluate the status of oxidative stress in a rat model of preeclampsia.. We measured the aortic AT(1) receptor expression and urinary excretion of 8-isoprostane (8IP) in rats rendered "preeclamptic" and compared the findings to those obtained in normal pregnant animals, pregnant rats injected with MBG, and preeclamptic rats treated with RBG.. Aortic AT(1) receptor expression and the urinary excretion of 8IP were significantly augmented in "preeclamptic" and MBG-injected pregnant rats compared to normal pregnant animals. RBG prevented evidence of oxidative stress in "preeclamptic" rats.. MBG is involved in the causation of oxidative stress in our rat model and RBG attenuates this change.

Topics: Animals; Bufanolides; Disease Models, Animal; Female; Oxidative Stress; Pre-Eclampsia; Pregnancy; Rats; Rats, Sprague-Dawley

Cardiotonic steroids (CTS) are implicated in pathophysiology of uremic cardiomyopathy. In the present study, we tested whether a monoclonal antibody (mAb) against the bufadienolide CTS, marinobufagenin (MBG), alleviates cardiac hypertrophy and fibrosis in partially nephrectomized (PNx) rats.. In PNx rats, we compared the effects of 3E9 anti-MBG mAb and of Digibind, an affinity-purified digoxin antibody, on blood pressure and cardiac hypertrophy and fibrosis following 4 weeks after the surgery.. In PNx rats, a fourfold elevation in plasma MBG levels was associated with hypertension, increased cardiac levels of carbonylated protein, cardiac hypertrophy, a reduction in cardiac expression of a nuclear transcription factor which is a negative regulator of collagen synthesis, Friend leukemia integration-1 (Fli-1), and an increase in the levels of collagen-1. A single intraperitoneal administration of 3E9 mAb to PNx rats reduced blood pressure by 59 mm Hg for 7 days and produced a significant reduction in cardiac weight and cardiac levels of oxidative stress, an increase in the expression of Fli-1, and a reduction in cardiac fibrosis. The effects of Digibind were similar to those of 3E9 mAb, but were less pronounced.. In experimental chronic renal failure, elevated levels of MBG contribute to hypertension and induce cardiac fibrosis via suppression of Fli-1, representing a potential target for therapy.

Topics: Animals; Antibodies, Monoclonal; Blood Pressure; Bufanolides; Cardiomegaly; Comorbidity; Disease Models, Animal; Fibrosis; Immunoglobulin Fab Fragments; Kidney Failure, Chronic; Male; Myocardium; Nephrectomy; Proto-Oncogene Protein c-fli-1; Rats; Rats, Sprague-Dawley

Preeclampsia (PE) is a disorder that results in significant fetomaternal morbidity and mortality with yet no definitive pharmacological intervention. It involves the development of de novo hypertension and proteinuria after 20 weeks of pregnancy. All too often, intrauterine growth restriction (IUGR) occurs. Evidence has accrued that implicates the cardiac glycosides (the cardenolides and the bufadienolides) as potentially involved in the pathophysiology of PE. These compounds act by inhibiting Na(+)/K(+) ATPase. Digibind (digoxin immune Fab) antagonizes this action of the cardenolides. It also has cross-reactivity against the bufadienolides, including marinobufagenin. This study investigated the effects of Digibind in a rat model of PE. We induced a syndrome in rats, which includes many of the phenotypic characteristics of human PE. Digibind, in escalating doses, was given on days 10 to 20 of pregnancy. Digibind produced significant lowering of the blood pressure and reduced proteinuria in our rat model of PE. However, it also did not avert IUGR. In view of these findings, in our experimental model of human PE, further studies in the quest for effective treatment of PE need to focus on pharmaceuticals that can remedy the syndrome without compromising the fetus.

Topics: Animals; Animals, Newborn; Blood Pressure Determination; Bufanolides; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Administration Schedule; Female; Fetal Growth Retardation; Immunoglobulin Fab Fragments; Injections, Intraperitoneal; Pre-Eclampsia; Pregnancy; Pregnancy, Animal; Probability; Proteinuria; Random Allocation; Rats; Reference Values; Risk Assessment; Treatment Outcome; Urinalysis

The bufodienolides are cardiac glycosides which have the ability to inhibit the enzyme, Na(+)/K(+) ATPase (sodium potassium adenosine triphosphatase). They are cardiac inotropes, cause vasoconstriction (and, potentially, hypertension) and are natriuretic. Evidence has accrued over time which supports the view that they are mechanistically involved in volume expansion-mediated hypertension. In this communication, the authors summarize data which support the view that the bufodienolides and, in particular, marinobufagenin (MBG) are involved in the pathogenesis of preeclampsia. In a rat model of the syndrome, MBG causes hypertension, proteinuria, intrauterine growth restriction and increased weight gain. All of these phenotypic characteristics are prevented by an antagonist to MBG, resibufogenin (RBG). The "preeclamptic" animals also develop a vascular leak syndrome, resulting in hemoconcentration. Abnormalities in the MAPK (mitogen-activated protein kinase) system play a role in the mechanism by which MBG produces the abnormalities in the pregnant rat. Studies to discover the relevance of these findings to human preeclampsia are currently underway in several laboratories and clinics.

Topics: Animals; Bufanolides; Disease Models, Animal; Female; Fetal Growth Retardation; Humans; Mitogen-Activated Protein Kinase Kinases; Pre-Eclampsia; Pregnancy; Proteinuria; Rats; Sodium-Potassium-Exchanging ATPase

Preeclampsia is a hypertensive disorder which develops de novo in women during pregnancy. The urinary excretion of the cardiotonic steroid, marinobufagenin (MBG), is increased prior to the development of hypertension. Preeclamptic patients are volume expanded but much of the excess salt and water appears to be located primarily in the interstitial space. Therefore, 'capillary leak' syndrome has been postulated in this disorder.. We evaluated the vascular leakage in normal rats following MBG injection and in a rat model of human preeclampsia. We measured the changes in light intensity comparing that in the intravascular to the extravascular space by assessing 'leak' of fluorescein-labeled albumin (FITC-albumin) from mesenteric postcapillary venules.. FITC-albumin extravasation continued to increase in a time-dependent fashion after MBG infusion and was significant (p < 0.05) at 60 min of observation when compared to sham rats. We also observed a significant difference in 'vascular leakage' in preeclamptic rats compared to control non-pregnant and normal pregnant groups starting at 20 min after the FITC-albumin infusion.. We propose that MBG is involved in the production of a 'vascular leak' in our rat model of preeclampsia.

Topics: Animals; Bufanolides; Caspase 3; Caspase 8; Disease Models, Animal; Female; Humans; Male; Pre-Eclampsia; Pregnancy; Pregnancy, Animal; Rats; Rats, Sprague-Dawley; Steroids; Vasoconstrictor Agents

Spironolactone has been noted to attenuate cardiac fibrosis. We have observed that the cardiotonic steroid marinobufagenin plays an important role in the diastolic dysfunction and cardiac fibrosis seen with experimental renal failure. We performed the following studies to determine whether and how spironolactone might ameliorate these changes. First, we studied rats subjected to partial nephrectomy or administration of exogenous marinobufagenin. We found that spironolactone (20 mg/kg per day) attenuated the diastolic dysfunction as assessed by ventricular pressure-volume loops and essentially eliminated cardiac fibrosis as assessed by trichrome staining and Western blot. Next, we examined the effects of spironolactone and its major metabolite, canrenone (both 100 nM), on marinobufagenin stimulation of rat cardiac fibroblasts. Both spironolactone and canrenone prevented the stimulation of collagen production by 1 nM marinobufagenin but not 100 nM marinobufagenin, as assessed by proline incorporation and procollagen 1 expression, as well as signaling through the sodium-potassium-ATPase, as evidenced by protein kinase C isoform delta translocation and extracellular signal regulated kinase 1/2 activation. Both spironolactone and canrenone also altered ouabain binding to cultured porcine cells in a manner consistent with competitive inhibition. Our data suggest that some of the antifibrotic effects of spironolactone may be attributed to antagonism of marinobufagenin signaling through the sodium-potassium-ATPase.

Topics: Animals; Bufanolides; Canrenone; Cardiomyopathies; Cardiotonic Agents; Cells, Cultured; Disease Models, Animal; Drug Interactions; Endomyocardial Fibrosis; Fibroblasts; Mineralocorticoid Receptor Antagonists; Myocardium; Nephrectomy; Ouabain; Procollagen; Proline; Rats; Renal Insufficiency; Spironolactone; Tritium; Uremia

The bufodienolides are natriuretic steroids, which also have the capacity to cause vasoconstriction, and are cardiac inotropes. Their mechanism of action appears to be related to their ability to inhibit Na+/K+ ATPase. The actions of one of these compounds, marinobufagenin (MBG), have been investigated in a rat model of preeclampsia, an example of volume expansion-mediated hypertension. The urinary excretion of MGB is increased in this model. Furthermore, this increment in its excretion occurs prior to the development of hypertension and proteinuria. The animals also demonstrate intrauterine growth restriction. Studies of the effect of MBG on cytotrophoblast cells reveal that MGB inhibits the migration, proliferation and invasion of these cells. We propose that MGB is an important etiologic factor in at least some forms of preeclampsia and that the level of its excretion in the urine may prove to be of diagnostic value.

Topics: Animals; Blood Vessels; Blood Volume; Bufanolides; Cardiac Glycosides; Disease Models, Animal; Female; Humans; Pre-Eclampsia; Pregnancy; Rats; Rats, Sprague-Dawley; Vasoconstrictor Agents

Levels of marinobufagenin (MBG), an endogenous bufadienolide Na/K-ATPase (NKA) inhibitor, increase in preeclampsia and in NaCl-sensitive hypertension.. We tested a 3E9 monoclonal anti-MBG antibody (mAb) for the ability to lower blood pressure (BP) in NaCl-sensitive hypertension and to reverse the preeclampsia-induced inhibition of erythrocyte NKA. Measurements of MBG were performed via immunoassay based on 4G4 anti-MBG mAb.. In hypertensive Dahl-S rats, intraperitoneal administration of 50 microg/kg 3E9 mAb lowered BP by 32 mmHg and activated the Na/K-pump in the thoracic aorta by 51%. NaCl supplementation of pregnant rats (n = 16) produced a 37 mmHg increase in BP, a 3.5-fold rise in MBG excretion, and a 25% inhibition of the Na/K-pump in the thoracic aorta, compared with pregnant rats on a normal NaCl intake. In eight pregnant hypertensive rats, 3E9 mAb reduced the BP (21 mmHg) and restored the vascular Na/K-pump. In 14 patients with preeclampsia (mean BP, 126 +/- 3 mmHg; 26.9 +/- 1.4 years; gestational age, 37 +/- 0.8 weeks), plasma MBG was increased three-fold and erythrocyte NKA was inhibited compared with that of 12 normotensive pregnant women (mean BP, 71 +/- 3 mmHg) (1.5 +/- 0.1 vs. 3.1 +/- 0.2 micromol Pi/ml/h, respectively; P < 0.01). Ex-vivo 3E9 mAb restored NKA activity in erythrocytes from patients with preeclampsia. As compared with 3E9 mAb, Digibind, an affinity-purified antidigoxin antibody, was less active with respect to lowering BP in both hypertensive models and to restoration of NKA from erythrocytes from patients with preeclampsia.. Anti-MBG mAbs may be a useful tool in studies of MBG in vitro and in vivo and may offer treatment of preeclampsia.

Topics: Adult; Animals; Antibodies, Monoclonal; Blood Pressure; Bufanolides; Digoxin; Disease Models, Animal; Female; Humans; Hypertension; Immunoglobulin Fab Fragments; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Third; Pregnancy, Animal; Rats; Rats, Inbred Dahl; Sensitivity and Specificity; Sodium Chloride, Dietary; Sodium-Potassium-Exchanging ATPase

There are two major pathophysiologic processes involved in the development of hypertension: (1) expanded extracellular fluid volume and (2) vasoconstriction. We have developed a model of preeclampsia in the rat, in which excessive volume expansion (VE) plays a role. These animals excrete increased amounts of the bufodienolide, marinobufagenin (MBG), even before their hypertension and proteinuria become established. Furthermore, their hypertension is corrected by administration of resibufogenin (RBG), a compound structurally similar to MBG.. We studied two models of experimental hypertension in the nonpregnant animal, produced either by deoxycorticosterone acetate (DOCA)-salt administration or by angiotensin infusion.. RBG administered to the DOCA-salt rats lowered blood pressure and reduced proteinuria in the VE animals, but had no affect on the rats infused with angiotensin. Furthermore, although the production of superoxide anion in the aortas of both groups of hypertensive rats was increased over control, RBG reduced these levels to normal in the VE (DOCA-salt) animals only. RBG had no effect in the angiotensin-infused rats. The urinary excretion of angiotensinogen did not rise in VE-mediated hypertension, but did increase in the angiotensin-infused rats.. MBG plays an important role in the causation of hypertension in the VE rats, but not in the vasoconstrictive model. RBG is effective only in VE-mediated hypertension.

Topics: Angiotensin II; Angiotensinogen; Animals; Blood Pressure; Bufanolides; Creatinine; Desoxycorticosterone; Disease Models, Animal; Hypertension, Renal; Male; Mineralocorticoids; Proteinuria; Rats; Rats, Inbred Strains; Sodium Chloride; Superoxides; Vasoconstrictor Agents

Because of the plethora of genetic manipulations available in the mouse, we performed a partial nephrectomy in the mouse and examined whether the phenotypical features of uremic cardiomyopathy described in humans and rats were also present in the murine model. A 5/6 nephrectomy was performed using a combination of electrocautory to decrease renal mass on the left kidney and right surgical nephrectomy. This procedure produced substantial and persistent hypertension as well as increases in circulating concentrations of marinobufagenin. Invasive physiological measurements of cardiac function demonstrated that the 5/6 nephrectomy resulted in impairment of both active and passive left ventricular relaxation at 4 wk whereas tissue Doppler imaging detected changes in diastolic function after 6 wk. Morphologically, hearts demonstrated enlargement and progressive fibrosis, and biochemical measurements demonstrated downregulation of the sarcoplasmic reticulum calcium ATPase as well as increases in collagen-1, fibronectin, and vimentin expression. Our results suggest that partial nephrectomy in the mouse establishes a model of uremic cardiomyopathy which shares phenotypical features with the rat model as well as patients with chronic renal failure.

Topics: Animals; Antihypertensive Agents; Blood Pressure; Bufanolides; Cardiomegaly; Cardiomyopathies; Disease Models, Animal; Echocardiography, Doppler; Extracellular Signal-Regulated MAP Kinases; Fibrosis; Heart; Male; Mice; Mice, Inbred Strains; Myocardium; Myocytes, Cardiac; Nephrectomy; Renal Insufficiency; Sarcoplasmic Reticulum Calcium-Transporting ATPases; Sodium-Potassium-Exchanging ATPase; src-Family Kinases; Stroke Volume; Ventricular Function, Left

The study of the pathogenesis of preeclampsia has been hampered by a relative dearth of animal models. We developed a rat model of preeclampsia in which the excretion of a circulating inhibitor of Na/K ATPase, marinobufagenin (MBG), is elevated. These animals develop hypertension, proteinuria, and intrauterine growth restriction. The administration of a congener of MBG, resibufogenin (RBG), reduces blood pressure to normal in these animals, as is the case when given to pregnant animals rendered hypertensive by the administration of MBG. Studies of Na/K ATPase inhibition by MBG and RBG reveal that these agents are equally effective as inhibitors of the enzyme.

Topics: Animals; Blood Pressure; Bufanolides; Disease Models, Animal; Enzyme Inhibitors; Female; Humans; Hypertension; Isoenzymes; Molecular Structure; Ouabain; Pre-Eclampsia; Pregnancy; Rats; Sodium-Potassium-Exchanging ATPase

Left ventricular hypertrophy commonly complicates chronic renal failure. We have observed that at least one pathway of left ventricular hypertrophy appears to involve signaling through reactive oxygen species (ROS). Green tea is a substance that appears to have substantial antioxidant activity, yet is safe and is currently widely used. We, therefore, studied whether green tea supplementation could attenuate the development of left ventricular hypertrophy in an animal model of chronic renal failure.. Male Sprague-Dawley rats were subjected to sham or remnant kidney surgery and given green tea extract (0.1% and 0.25%) or plain drinking water for the next 4 weeks. Heart weight, body weight, and cardiac Na-K-ATPase activity were measured at the end of this period. To further test our hypothesis, we performed studies in cardiac myocytes isolated from adult male Sprague-Dawley rats. We measured the generation of ROS using the oxidant sensitive dye dichlorofluorescein (DCF) as well as (3H)phenylalanine incorporation following exposure to cardiac glycosides with and without green tea extract.. Administration of green tea extract at 0.25% resulted in attenuation of left ventricular hypertrophy, hypertension, and preserved cardiac Na-K-ATPase activity in rats subjected to remnant kidney surgery (all P < 0.01). In subsequent studies performed in isolated cardiac myocytes, both ouabain and marinobufagenin (MBG) were both found to increase ROS production and (3H)phenylalanine incorporation at concentrations substantially below their inhibitor concentration (IC) 50 for the sodium pump. Addition of green tea extract prevented increases in ROS production as well as (3H)phenylalanine incorporation in these isolated cardiac myocytes.. Green tea extract appears to block the development of cardiac hypertrophy in experimental renal failure. Some of this effect may be related to the attenuation of hypertension, but a direct effect on cardiac myocyte ROS production and growth was also identified. Clinical studies of green tea extract in chronic renal failure patients may be warranted.

Topics: Animals; Blood Pressure; Bufanolides; Cardenolides; Cell Division; Cells, Cultured; Digoxin; Disease Models, Animal; Enzyme Activation; Hypertrophy, Left Ventricular; Kidney Failure, Chronic; Male; Myocardium; Myocytes, Cardiac; Nephrectomy; Phytotherapy; Plant Extracts; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Rubidium Radioisotopes; Saponins; Sodium-Potassium-Exchanging ATPase; Tea