Obesity affects short-term folate pharmacokinetics in women of childbearing age.

Maternal folate status and body mass index (BMI) are independent risk factors for neural tube defects (NTD). Population-based studies have identified an inverse association between serum folate and BMI, after adjusting for intake. The objective of this intervention study was to compare the relationship between BMI and the short-term pharmacokinetic response to an oral dose of folic acid. Healthy obese (BMI 30.0 kg m(-2); n=16) and normal-weight (BMI 18.5-24.9 kg m(-2); n=16) women of childbearing age (18-35 years) were administered a single oral dose of folic acid (400 µg). Blood samples were collected over a 10-h period to evaluate the serum folate response. Fasting baseline serum folate was lower in the obese group (P=0.005); in contrast, red blood cell folate was higher (P=0.05). Area-under-the-curve for the absorption phase (0-3 h) and peak serum folate concentrations were lower in obese versus normal-weight women (P<0.005). Overall serum folate response (0-10 h) was lower in obese versus normal-weight women (repeated-measures ANOVA, P=0.001). Data suggest body distribution of folate is significantly affected by obesity, and, should pregnancy occur, may reduce the amount of folate available to the developing embryo. These findings provide additional support for a BMI-adjusted folic acid intake recommendation for NTD risk reduction.

Effect of St. John's wort on free radical production.

St. John's wort (Hypericum perforatum) is an herbal compound used in the treatment of burns, bruises, swelling, anxiety, and most recently, mild to moderate depression. The present study was designed to evaluate the antioxidant properties of St. John's wort in both cell-free and human vascular tissue. The experiment was performed initially in a cell-free system using Krebs buffer and a combination of xanthine/xanthine oxidase to initiate the production of the superoxide radical. Additionally, human placental vein was incubated in Krebs buffer without xanthine or xanthine oxidase to study the effects of St. John's wort on human tissue in vitro. Commercially available formulations of St. John's wort, standardized to either hypericin or hyperforin, were dissolved in an alkaline solution, and the following dilutions were made: 1:1, 1:2.5, 1:5, 1:7.5, 1:10, and 1:20. Lucigenin chemiluminescence was used to measure free radical production in both systems. A pro-oxidant effect was seen at the highest concentration, 1:1. Lower concentrations revealed antioxidant properties of the compound. All dilutions below 1:1 in both systems showed a dose-related inverse relationship of superoxide inhibition. The largest suppression was seen at the most dilute concentration, 1:20. The addition of 10(-3) M tiron inhibited the chemiluminescence signal, thereby confirming the production of superoxide. The results of this study suggest that St. John's wort inhibits free radical production in both cell-free and human vascular tissue.

Determination of arginine and methylated arginines in human plasma by liquid chromatography-tandem mass spectrometry.

Nitric oxide (NO) is synthesized from L-arginine (ARG) catalyzed by the enzyme nitric oxide synthase (NOS) and is important in the regulation of vascular tone, neurotransmission and host defense. N,N-Dimethyl L-arginine (asymmetric dimethylarginine, ADMA) and N-monomethyl L-arginine (MMA) are endogenous inhibitors of NOS. N,N'-Dimethyl L-arginine (symmetric dimethylarginine, SDMA), the inactive enantiomer of ADMA is also known to be present endogenously. A simple, sensitive and fast LC-MS-MS method was developed to extract and quantitate ADMA, SDMA, MMA and ARG from human plasma. 13C6-ARG was used as the internal standard for the assay. Protein precipitation using acetonitrile gave good recoveries of all the compounds from plasma. The compounds were separated by HPLC in less than 15 min using a silica column. The limits of detection for this method were found to be approximately 1 ng/ml for ARG, ADMA and SDMA and 2.5 ng/ml for MMA. The total LC-MS-MS analysis time is less than 15 min making this the fastest and most specific method reported to date. The use of an isocratic liquid chromatographic separation makes this method optimal for high sample throughput. The inter- and intra-day precision (% RSD) and accuracy (% error) for this assay were less than 15%. The average concentrations of ARG, ADMA, SDMA and MMA in plasma from 20 human subjects were found to be 10.9+/-4.1 microg/ml, 25.1+/-9.4 ng/ml, 33.2+/-13.1 ng/ml and 19.6+/-3.8 ng/ml, respectively.

Distribution of endothelin receptors in saphenous veins of African Americans: implications of racial differences.

The relative density of endothelin-receptor subtypes A and B (ET(A) and ET(B), respectively) on endothelial and smooth muscle cells is the major determinant of the contractile response to endothelin-1 (ET-1). To investigate the effects of race on the distribution of ET receptors, the endothelin-receptor subtypes on membrane fractions prepared from saphenous veins obtained from African-American patients undergoing coronary bypass surgery were analyzed. Similar studies were repeated with endothelium-denuded samples to study the role of endothelium- and smooth muscle-derived ET(B) receptors. Competitive-binding experiments on membrane fractions by using [125I]-ET-1 and unlabeled ligands, ET-1, ET-3, sarafotoxin-6-c, and BQ-123 yielded two classes of binding sites on endothelium-intact vessels from both female and male subjects. In women, the maximal binding capacities were 91+/-6 and 67+/-13 fmol/mg protein for ET(A) and ET(B) receptors, respectively; the corresponding values in men were 178+/-19 and 127+/-13 fmol/mg protein. Similar experiments performed with endothelium-denuded saphenous veins indicated the presence of both receptor subtypes on vascular smooth muscle, in contrast to our earlier report on the presence of only ET(A) receptors on vessels obtained from white Americans. Our findings demonstrate that the ratio and the density of ET receptors are different in black and white Americans.

St. John's wort: a new alternative for depression?

OBJECTIVE: The primary purpose of this article is to review the existing literature concerning the therapeutic uses, adverse effects, and possible drug interactions of St. John's wort (Hypericum perforatum) as compared to other antidepressant medications. METHODS: Reference material was obtained through database searches with time restrictions of 1985 to the present. Studies selected were those written in the English language which compared the role of St. John's wort, tricyclic antidepressants, monoamine oxidase inhibitors, and serotonin-selective reuptake inhibitors in the treatment of depression. Other studies were selected based on their evaluation of the safety and efficacy of St. John's wort as an antidepressant for a minimum of four weeks. RESULTS: A review of existing literature recognized nine clinical trials that reported the efficacy of St. John's wort as compared to placebo and to other antidepressant medications. Of these nine, four controlled studies were chosen based upon their large patient populations and their consistency in brand and dosage of St. John's wort used. These four studies demonstrated that St. John's wort was as effective as other antidepressant medications and more effective than placebo, as the clinical symptoms of depression greatly decreased upon administration of H. perforatum. The side-effect profile of H. perforatum at this time appears to be superior to any current U.S.-approved antidepressant medication. CONCLUSIONS: From the existing literature, St. John's wort appears to be a safe and effective alternative in the treatment of depression. Tricylic antidepressants and monoamine oxidase inhibitors can produce serious cardiac side-effects, such as tachycardia and postural hypotension, and many unwanted anticholinergic side-effects, including dry mouth and constipation. St. John's wort has proven to be free of any cardiac, as well as anticholinergic, side-effects normally seen with antidepressant medications. Based upon limited studies, St. John's wort appears to be an acceptable alternative to traditional antidepressant therapy, although trials on a larger scale are warranted in this area. Hypericum is available to the lay public as an over-the-counter preparation and may be misused if not fully understood.

Palmar digital vessel relaxation in healthy horses and in horses given carbohydrate.

OBJECTIVE: To compare in vitro smooth muscle relaxation of palmar digital vessels from healthy horses with those from horses in the prodromal stage of experimentally (carbohydrate) induced laminitis. ANIMALS: 16 adult horses. PROCEDURE: Segments of palmar digital vessels were obtained from 5 healthy horses and 6 horses given carbohydrate. Vascular rings from the palmar digital artery and vein were suspended in individual organ baths containing buffer solution and indomethacin; isometric tension was recorded, and contraction and relaxation were compared. Smooth muscle contraction in response to cumulative addition of phenylephrine was recorded in the absence and presence of 1 microM NG-nitro-L-arginine methyl ester (L -NAME). After wash out, vascular rings were preconstricted with phenylephrine (0.3 microM), and cumulative endothelium-dependent (acetylcholine-induced) and independent (nitroprusside-induced) smooth muscle relaxations were recorded in the absence or presence of L -NAME. RESULTS: Phenylephrine increased vascular smooth muscle tone in ring preparations of palmar digital arteries and veins. Addition of acetylcholine or nitroprusside induced relaxation of palmar digital artery and vein ring preparations. Use of L-NAME (1 microM) significantly reduced maximal relaxation induced by acetylcholine, but not by nitroprusside. Maximal relaxation induced by acetylcholine, but not by nitroprusside, was reduced in vascular rings prepared from carbohydrate-overloaded horses. CONCLUSION AND CLINICAL RELEVANCE: Reduced endothelium-dependent relaxation of palmar digital vessels may have a role in the pathophysiology of acute laminitis after carbohydrate overload in horses.

Enhanced hemodynamic response to [D-ALA2,D-MET5]-methionine enkephalin (DAME) in streptozotocin-induced diabetic rats is reversed by insulin replacement.

The present study examined the alterations of hemodynamic responses to [D-ala2,D-Met5]-methionine enkephalin (DAME) in diabetic animals. Male Sprague-Dawley rats (12 weeks old) were used for this study. Diabetes was induced by a single injection of streptozotocin (65 mg/kg, i.v.). After 7 days, blood glucose levels were determined to confirm the diabetic state. Animals were anesthetized and instrumented to monitor mean arterial pressure, hindlimb bloodflow and hindlimb vascular resistance. Administration of DAME produced a significantly greater reduction in blood pressure, increase in hindlimb bloodflow and decrease in hindlimb vascular resistance in diabetic vs. control rats. These effects were blocked by naloxone. All hemodynamic changes were attenuated after pretreatment with the ganglionic blocker, hexamethonium, indicating that the responses were mediated either within the central nervous system or at the ganglia. Insulin reversed the exaggerated depressor effect of DAME on streptozotocin-treated rats. Collectively, these results suggest that diabetic rats have altered opioidergic hemodynamic responses to DAME due to mu receptor alterations in the CNS or in autonomic ganglia. These effects were reversed by replacement of insulin.

Gender differences in the expression of endothelin receptors in human saphenous veins in vitro.

The contractile response to endothelin-1 (ET-1) appears to be modulated by the relative density of ETA and ETB receptors. To determine the effects of gender on the distribution of ET receptors, we analyzed the endothelin receptor subtypes on membrane fractions prepared from saphenous vein samples obtained from patients of different genders undergoing coronary artery bypass graft surgery. The contractile response to ET-1 in the presence and absence of 1 microM of the ETA receptor antagonist BQ-123 was also investigated. Similar studies were repeated with endothelium-denuded samples to study the role of endothelium- and smooth muscle-derived ETB receptors. Competitive binding experiments were performed on membrane fractions using [125I]ET-1 and unlabeled ligands ET-1, ET-3, sarafatoxin 6c and BQ-123. Analysis of the binding data with endothelium-intact samples yielded two classes of binding sites in both women and men. In women, the maximum binding capacities were 83 +/- 6 and 97 +/- 10 fmol/mg protein for ETA and ETB receptors, respectively; the corresponding values in men were 618 +/- 121 and 201 +/- 10 fmol/mg protein. In addition, ET-1-induced contractions were 2-fold greater in men than in women at high ET-1 concentrations. Competitive binding studies with endothelium-denuded saphenous veins demonstrated the presence of only ETA receptors in both female and male tissue. These results indicate that the ratio and the density of ET receptors are different in men and women, which might be an important factor in the regulation of the contractile response.

Oxidized low-density lipoprotein inhibits acetylcholine-induced vasorelaxation and increases 5-HT-induced vasoconstriction in isolated human saphenous vein.

The present study determined the vasomotor effects of oxidized low-density lipoprotein (ox-LDL) in human saphenous veins and determined whether decreased availability of L-arginine was responsible for the impaired endothelial function. Human saphenous veins were obtained from white males undergoing coronary bypass surgery. We examined the effects of ox-LDL on ACh-induced endothelium-dependent relaxation, sodium nitroprusside-induced endothelium-independent relaxation and 5-HT-induced contraction. ACh-induced vasorelaxation in the presence of L-arginine and ox-LDL was also examined. In addition, we assessed the endothelial influence on the contractile response to 5-HT. ox-LDL significantly inhibited ACh-induced relaxation but did not affect sodium nitroprusside-induced relaxation. L-Arginine pretreatment did not prevent ox-LDL-induced impairment of the relaxation response to ACh. ox-LDL significantly potentiated 5-HT-induced contraction at concentrations between 3 x 10(-6) M and 10(-4) M, an effect that was endothelium-dependent. Denudation of endothelium also significantly enhanced the contractile response to 5-HT. These data suggest that ox-LDL impairs ACh-induced endothelium-dependent relaxation and enhances 5-HT-induced endothelium-dependent contraction in human saphenous vein. L-Arginine deficiency is not responsible for the endothelial dysfunction induced by ox-LDL in human saphenous vein.

Indicators of postoperative pain in cats and correlation with clinical criteria.

OBJECTIVE: To identify clinical indicators that may help identity postoperative pain in cats after ovariohysterectomy. ANIMALS: Healthy, laboratory animal source cats. PROCEDURE: Clinical indicators of pain were identified, and relief from pain in response to butorphanol was studied in 5 groups of cats. 10 cats had 1 hour of general anesthesia only, followed by recovery without additional medication. 10 cats had general anesthesia and ovariohysterectomy, followed by recovery without additional medication. 10 cats had general anesthesia, ovariohysterectomy, and postoperative administration of 0.1 mg of butorphanol/kg of body weight. Another 10 cats had general anesthesia, ovariohysterectomy, and postoperative administration of 0.3 mg butorphanol/kg. 10 cats received 0.1 mg of butorphanol/kg, IM, only. Samples and recorded data were obtained before, during, and after the anesthesia period. Clinical variables measured included heart rate, blood pressure, respiratory rate, rectal temperature, PCV, and blood glucose concentration. Results were compared with changes in norepinephrine, epinephrine, and cortisol concentrations. RESULTS: Cats that did not receive analgesics had higher cortisol concentration than did cats without surgery and cats that received butorphanol after surgery. Systolic blood pressure measured by ultrasonic Doppler was found to be predictive of cortisol concentration, using a multiple linear regression model. CONCLUSIONS: Cortisol concentration increased in response to surgical stress and pain, and this increase was diminished by use of butorphanol. CLINICAL RELEVANCE: Systolic blood pressure was the best clinical predictor of postoperative pain.

Identification of receptor binding and activation sites in endothelin-1 by use of site-directed mutagenesis.

This study addresses the structural requirements for the intracellular processing and receptor binding properties of endothelin-1 (ET-1). Point mutants of preproendothelin-1 cDNA, with replacement of the codons for Lys9 of ET-1 by ones for Ala and Glu and of Ile20 and Trp21 by ones encoding Ala, were expressed in COS-7 cells. Competitive binding experiments on rat vascular smooth muscle cells (A-10), which were shown to be an ETA receptor-rich cell line, between [125I]ET-1 and synthetic ET-1, wild-type recombinant ET-1, and recombinant [Ala9]ET-1, [Glu9]ET-1, [Ala20]ET-1, and [Ala21]ET-1 yielded Ki values of 0.2 +/- 0.02, 0.2 +/- 0.02, 0.04 +/- 0.01, 1.4 +/- 0.2, 1.6 +/- 0.2, and > 50 nmol/L, respectively. In similar experiments with ETB receptor-rich human Girardi heart cells, the corresponding values were 0.2 +/- 0.03, 0.2 +/- 0.03, 0.2 +/- 0.04, 0.2 +/- 0.06, 1.4 +/- 0.4, and > 50 nmol/L. The ETA receptor-mediated contractile responses to [Glu9]ET-1 and [Ala20]ET-1, measured by using canine coronary artery rings, were decreased approximately fourfold to fivefold compared with the response produced by synthetic or wild-type recombinant ET-1, whereas [Ala9]ET-1 was found to be more potent, and [Ala21]ET-1 did not produce any contraction. These results demonstrate that Ile20 and Trp21 are involved in binding to both receptor subtypes. Of considerable interest was the observation that [Glu9]ET-1 also blunts the ETA receptor subtype-mediated contractile response to ET-1 stimulus.

Insulin attenuates alpha-adrenergic aortic contraction in normotensive but not borderline hypertensive rats.

Hyperinsulinemia can alter vasoconstrictor responses in normotensive and hypertensive rats, but the effects of insulin on vascular contraction have not been evaluated in borderline hypertension. This study determined the effects of insulin on alpha-adrenoceptor mediated aortic contraction in male and female borderline hypertensive rats (BHR) and normotensive Wistar Kyoto (WKY) rats. Dose-response curves to norepinephrine (NE) and phenylephrine (PE) were performed in thoracic aorta in the absence or presence of insulin (100 microU/ml) for 2 hrs. Contraction to NE and PE was reduced in aorta from female WKY rats incubated with insulin compared to control. In aorta from 6 of 13 male WKY rats (insulin responders), an attenuated response to NE and PE was observed in the presence of insulin. However, insulin did not alter responses to NE or PE in aorta from male or female BHR. These results indicate that insulin impairs alpha-adrenoceptor mediated contraction in the normotensive female WKY rat and in a group of responder male WKY rats, but not in BHR. This study supports the presence of a resistance to the vascular effects of insulin in BHR.

Physiologically based pharmacokinetic model useful in prediction of the influence of species, dose, and exposure route on perchloroethylene pharmacokinetics.

The ability of a physiologically based pharmacokinetic (PBPK) model to predict the uptake and elimination of perchloroethylene (PCE) in venous blood was evaluated by comparison of model simulations with experimental data for two species, two routes of exposure, and three dosage levels. Unanesthetized male Sprague-Dawley rats and beagle dogs were administered 1, 3, or 10 mg PCE/kg body weight in polyethylene glycol 400 as a single bolus, either by gavage or by intraarterial (ia) injection. Serial blood samples were obtained from a jugular vein cannula for up to 96 h following dosing. The PCE concentrations were analyzed by headspace gas chromatography. For each dose and route of administration, terminal elimination half-lives in rats were shorter than in dogs, and areas under the blood concentration-time curve were smaller in rats than in dogs. Over a 10-fold range of doses, PCE blood levels in the rat were well predicted by the PBPK model following ia administration, and slightly underpredicted following oral administration. The PCE concentrations in dog blood were generally overpredicted, except for fairly precise predictions for the 3 mg/kg oral dose. These studies provide experimental evidence of the utility of the PBPK model for PCE in interspecies, route-to-route, and dose extrapolations.

Nonsuperoxide lucigenin-enhanced chemiluminescence from phospholipids and human saphenous veins.

The use of lucigenin-enhanced chemiluminescence (CL) for the detection of superoxide (O2.-) has grown in popularity due to an increased demand for a simple and specific system capable of measuring superoxide. In this study we report a lucigenin-CL signal emanating from human saphenous veins (SV) that was not inhibited by superoxide dismutase (SOD) and lasted for more than 24 h. A larger CL-signal with similar properties was produced by saphenous veins that had been dehydrated. A similar, non-SOD-inhibitable lucigenin-CL was also produced with a variety of phospholipids and phosphatidic acid. The chemical moiety responsible for the phospholipid CL is oxygen dependent but remains unidentified because a variety of lipids and phosphate containing species failed to produce such a signal. These results suggest that the use of lucigenin as a specific CL enhancer for O2.- must be clearly discriminated with a specific O2.- inhibitor when used in biological systems.

The use of methylene blue as an extravascular surgical marker impairs vascular responses of human saphenous veins.

Methylene blue is occasionally applied to the adventitia of blood vessels during coronary artery bypass and other vascular procedures to assist in the orientation of the vessel. Inherent in this method is the assumption that extravascular application of methylene blue is innocuous with regard to vascular function. In the first part of this study, the in vitro vascular reactivity of methylene blue-labeled saphenous veins was compared with that of veins that were not marked with methylene blue. The vasoactive agents tested were designed to examine multiple pathways. They included potassium chloride, prostaglandin F2 alpha, phenylephrine, serotonin, angiotensin II, BHT-933 (alpha 2-adrenergic agonist), sodium nitroprusside, acetylcholine, isoproterenol, and verapamil. Compared with unmarked veins, those marked with methylene blue demonstrated a significant impairment of both vasoconstrictor and vasodilator function. These observations were made on a relatively small number of patients and could therefore be attributed to inherent differences between patients or surgical procedures. In the second part of this study, these variables were eliminated by dividing a single vein from one patient into three segments for a 45-minute exposure to external only methylene blue, internal and external methylene blue, or no methylene blue. The segments were then evaluated for vasoreactivity in vitro. Externally applied methylene blue reduced vasoconstriction regardless of the agonist. Further, both endothelium-dependent and -independent vasodilation was diminished by external methylene blue exposure. In veins exposed to methylene blue both internally and externally the results were similar but the magnitude of impairment greater. It is concluded that surgical marking of blood vessels with methylene blue has the potential to adversely affect vascular reactivity and therefore the use of alternative dyes should be considered.

Opioidergic receptors in the arcuate nucleus are not involved in the cardiovascular effects of clonidine.

The arcuate nucleus is the bed nucleus for the pro-opiomelancortin system of the brain with important connections with other nuclei involved in cardiovascular function. Clonidine has been reported to produce its cardiovascular effects through an interaction with opioid and alpha 2-adrenergic receptors. The present study examined the arcuate nucleus as a site of action of clonidine. Male spontaneously hypertensive rats were anesthetized with pentobarbital and were instrumented for the measurement of blood pressure and heart rate. Cannulae were placed either through the cisterna magna (IC) or in the arcuate nucleus. Administration of clonidine (0.03-3.75 micrograms, IC) produced a dose-dependent hypotension and bradycardia. Pretreatment with naloxone (30 micrograms, IC) prior to clonidine administration resulted in a significant attenuation of both the clonidine-induced hypotension and bradycardia. In contrast, administration of naloxone (100 ng) into the arcuate nucleus prior to the central administration of clonidine did not alter the cardiovascular effects of clonidine. These results support the role of central opioidergic receptors in the cardiovascular effects of clonidine but do not support the arcuate nucleus as the site of action.

Use of tissue disposition data from rats and dogs to determine species differences in input parameters for a physiological model for perchloroethylene.

Tissue disposition of perchloroethylene (PCE) was determined experimentally in two mammalian species of markedly different size in order to derive input parameters for the development of a physiologically based pharmacokinetic (PBPK) model, which could forecast the disposition of PCE in each species. Male Sprague-Dawley rats and male beagle dogs received a single bolus of 10 mg PCE/kg body wt in polyethylene glycol 400 by gavage. Serial samples of brain, liver, kidney, lung, heart, skeletal muscle, perirenal fat, and blood were taken for up to 72 hr following PCE administration. Blood and tissue PCE concentrations were analyzed using a gas chromatography headspace technique. Dogs exhibited considerably longer tissue and blood half-lives than did rats. The dogs also exhibited larger area under tissue concentration versus time curves for all tissues except the liver. Whole body clearance of PCE in the rat was greater than that in the dog. Model simulations indicated this could be attributed to more rapid and extensive PCE exhalation and metabolism by the rat. The in vivo blood:air partition coefficient determined for rats was similar to an in vitro value previously reported. In vivo tissue: blood partition coefficients, however, were 1.4 to 2.8 times greater than published in vitro values. The PCE in vivo blood:air partition coefficient for the dog was twice that of the rat, but tissue:blood partition coefficients were 1.5 to 3.0 times greater in the rat than in the dog. These results demonstrated the existence of significant differences in partition coefficients in two species commonly used in toxicity testing. The PBPK model was shown to have utility in predicting the impact of metabolism and exhalation on pharmacokinetics of PCE in different species of widely differing size.

Copper deficient rat heart can compensate for doxorubicin-induced oxidant stress.

The hypothesis that copper (Cu) alters drug metabolizing enzymes and functions as an antioxidant nutrient in doxorubicin cardiotoxicity was tested. Male Sprague-Dawley rats were fed Cu adequate (+Cu; 5 mg Cu/kg of diet), marginally Cu deficient (MCu; 1.2 mg Cu/kg of diet), or severely Cu deficient (-Cu; 0.5 mg Cu/kg of diet) diets for 6 wk. Doxorubicin (1, 2, or 4 mg/kg body wt) or saline were administered intraperitoneally 1 time/wk for 4 wk. Compared to control hearts, Cu,Zn superoxide dismutase activity was decreased by 9% in MCu rats and by 21-40% in -Cu rats. Glutathione peroxidase activity was elevated 5-15% in -Cu rats. Doxorubicin administration increased heart Cu,Zn superoxide dismutase activity in +Cu and -Cu rats 18 h after the last of 4 injections, but not 18 h after 1 injection. There was no synergism between doxorubicin and Cu deficiency on lipid peroxidation, plasma creatine phosphokinase, cardiac hypertrophy, electrocardiographic abnormalities, or morphological changes. Heart glutathione S-transferase activity was decreased by Cu deficiency, and like Cu,Zn superoxide dismutase activity, returned to normal in -Cu rats given doxorubicin. Thus, the Cu deficient rat heart may be able to compensate for doxorubicin-induced oxidant stress by increasing the activity of Cu,Zn superoxide dismutase and glutathione S-transferase.

Copper and selenium deficiencies do not enhance the cardiotoxicity in rats due to chronic doxorubicin treatment.

This study tests the hypothesis that Cu and Se deficiencies enhance doxorubicin-induced cardiotoxicity and anemia. Male Sprague-Dawley rats (n = 48) were fed Cu and Se-adequate (+Cu+Se), Cu-deficient (-Cu), Se-deficient (-Se) or Cu and Se-deficient (-Cu-Se) diets for 5.5 wk. Doxorubicin (4 mg/kg body wt) or saline was administered once weekly for the last 4 wk of the study. Copper deficiency was confirmed by 79% lower liver Cu, 67% lower liver Cu,Zn superoxide dismutase (Cu,Zn SOD) activity and 76% lower erythrocyte Cu,Zn SOD activity. Selenium deficiency was confirmed by 90% lower liver glutathione peroxidase activity. Rats fed the -Cu diet had greater reductions in hematocrit than did those fed the +Cu diet after administration of doxorubicin. Doxorubicin, Cu deficiency and Se deficiency all produced electrocardiographic abnormalities and ultrastructural anatomical lesions. However, the dietary deficiencies did not enhance doxorubicin-induced cardiotoxicity. Doxorubicin, but not Cu or Se deficiency, raised lipid peroxidation 16% in liver (P < 0.01) and 18% in heart (not significant). These data suggest that the cardiomyopathies caused by doxorubicin and Cu and Se deficiencies have some similarities, but cardiac changes may be related to mechanisms other than lipid peroxidation.