Neuroprotective Effectiveness of Intravenous Ubiquinone in Rat Model of Irreversible Cerebral Ischemia.

The neuroprotective effect of ubiquinone (coenzyme Q10)was demonstrated on the rats model of ischemic stroke provoked by persistent 24-h occlusion of the middle cerebral artery. Coenzyme Q10 (30 mg/kg) was injected intravenously in 60 min after artery occlusion. Ubiquinone crossed the blood-brain barrier, accumulated in the brain, and produced a neuroprotective effect: it alleviated ischemia-induced neurological deficit and reduced the size of necrotic zone by 49% in comparison with rats receiving physiological saline.

[COMPARATIVE CARDIOPROTECTIVE EFFICACY OF COENZYME Q10 AND MEXICOR IN EXPERIMENTAL MODEL OF MYOCARDIAL INFARCTION IN RATS].

Cardioprotective efficacy of coenzyme Q10 (CoQ10, ubidecarenone) and mexicor were evaluated on the 21st day of experimental myocardial infarction in Wistar rats. CoQ10 or mexicor were injected in a dose of 30 mg/kg intravenously 10 min after coronary artery occlusion. The observed cardioprotective effects of ubidecarenone and mexicor were close. Both drugs equally increased the survival of rats, prevented the development of dilatation and hypertrophy of the left ventricle, and improved the pump cardiac function.

[Hplc estimation of coenzyme Q(10) redox status in plasma after intravenous coenzyme Q(10) administration].

The pharmacokinetics of the total pool of coenzyme Q(10) (Co(10)), its oxidized (ubiquinone) and reduced (ubiquinol, CoQ(10)H2) forms have been investigated in rats plasma during 48 h after a single intravenous injection of a solution of solubilized CoQ(10) (10 mg/kg) to rats. Plasma levels of CoQ(10) were determined by HPLC with spectrophotometric and coulometric detection. In plasma samples taken during the first minutes after the CoQ(10) intravenous injection, the total pool of coenzyme Q(10) and proportion of CoQ(10)H2 remained unchanged during two weeks of storage at -20°C. The kinetic curve of the total pool of coenzyme Q(10) corresponds to a one-part model (R² = 0.9932), while the corresponding curve of its oxidized form fits to the two-part model. During the first minutes after the injection a significant portion of plasma ubiquinone undergoes reduction, and after 7 h the concentration of ubiquinol predominates. The decrease in the total plasma coenzyme Q(10) content was accompanied by the gradual increase in plasma ubiquinol, which represented about 90% of total plasma CoQ(10) by the end of the first day. The results of this study demonstrate the ability of the organism to transform high concentrations of the oxidized form of CoQ(10) into the effective antioxidant (reduced) form and justify prospects of the development of parenteral dosage forms of CoQ(10) for the use in the treatment of acute pathological conditions.

Single intravenous injection of coenzyme Q10 protects the myocardium after irreversible ischemia.

Experiments were performed on the model of irreversible myocardial ischemia in Wistar rats. Coenzyme Q10 was injected intravenously 10 min after coronary artery occlusion. On day 21 after myocardial infarction the content of coenzyme Q10 in the left ventricle, liver, and plasma from animals of the treatment group was higher than that in untreated rats by 23, 1042, and 87%, respectively (p<0.05). The area of the necrotic zone was lower, and postinfarction hypertrophy of the left ventricle was less pronounced in coenzyme-receiving rats. Right ventricular hypertrophy did not develop in these animals. These rats were characterized by greater stroke volume (by 24.6%, p<0.05), stroke work (by 34.9%), cardiac output (by 37.8%, p<0.05), ejection fraction (by 35.7%, p<0.05), and contractility (by 22.5%, p<0.05), but lower end-diastolic pressure (by 25.8%, p<0.05) than untreated animals. These data indicate that the development of parenteral ubiquinone preparations holds much promise for urgent therapy of acute cardiovascular disorders.

Pharmacokinetics of coenzyme q10.

The pharmacokinetics of coenzyme Q10 powder and solution of solubilized form was studied after their oral administration to rats (10 mg/kg). Plasma concentrations of coenzyme Q10 were measured by HPLC with electrochemical detection over 48 hours. Solubilized coenzyme Q10 exhibited high absorption creating higher plasma concentrations of the drug, as a result of which its bioavailability constituted 264% of that for the powder.

Chronic administration of coenzyme Q10 limits postinfarct myocardial remodeling in rats.

The effect of chronic coronary artery occlusion on the content of rat myocardial coenzymes Q (CoQ) and evaluation of the applicability of CoQ(10) for limiting postinfarct remodeling have been investigated. Left ventricle myocardium hypertrophy was characterized by the decrease in CoQ(9) (-45%, p < 0.0001), CoQ(10) (-43%, p < 0.001), and alpha-tocopherol (-35%, p < 0.05). There were no differences between the parameters of postinfarction and sham-operated rats in plasma. Administration of CoQ(10) (10 mg/kg) via a gastric probe for 3 weeks before and 3 weeks after occlusion maintained higher levels of CoQ in the postinfarction myocardium: the decrease in CoQ(9) and CoQ(10) was 25% (p < 0.05) and 23% (p < 0.05), respectively (versus sham-operated animals). Plasma concentrations of CoQ(10) were more than 2 times higher (p < 0.05). In CoQ treated rats there was significant correlation between plasma levels of CoQ and the infarct size: r = -0.723 (p < 0.05) and r = -0.839 (p < 0.01) for CoQ(9) and CoQ(10). These animals were also characterized by earlier and more intensive scar tissue formation in the postinfarction myocardium and also by more pronounced cell regeneration processes. This resulted in the decrease in both the infarct size (16.2 +/- 8.1 vs. 27.8 +/- 12.1%) and also mass index of left ventricle (2.18 +/- 0.24 vs. 2.38 +/- 0.27 g/kg) versus untreated rats (p < 0.05). Thus, long-term treatment with ubiquinone increases plasma and myocardial CoQ content and this can improve the survival of myocardial cells during ischemia and limit postinfarct myocardial remodeling.

Role of reactive oxygen species in the sensitivity of rat hypertrophied myocardium to ischemia.

The relationship between hydroxyl radical (OH*) generation in the zone of ischemia/reperfusion and the size of infarction formed was investigated in 18-22-week-old anaesthetized male SHRSP and Wistar rats using a myocardial microdialysis technique. The marker of OH* generation, 2,3-dihydroxybenzoic acid (2,3-DHBA), was analyzed in dialyzates by high performance liquid chromatography with electrochemical detection. Myocardial ischemia was induced by ligation of the descending branch of the left main coronary artery for 30 min. The mean value of basal 2,3-DHBA level in the dialyzate samples from SHRSP (243 +/- 21 pg for 30 min) was significantly higher than that from Wistar rats (91 +/- 4 pg for 30 min, p < 0.0002); it positively correlated with left ventricular hypertrophy (r = 0.806; p < 0.05). During reperfusion total 2,3-DHBA output was 1.8-fold higher in SHRSP than in Wistar rats (659 +/- 60 pg versus 364 +/- 66 pg for 60 min, respectively, p < 0.0002). At the same time, 2,3-DHBA increase above the basal level was the same in Wistar and SHRSP rats (181 +/- 25 and 172 +/- 36 pg for 60 min, respectively). The infarct size in SHRSP (45.4 +/- 4.3%) was significantly higher (p < 0.05) than in Wistar rats (32.8 +/- 3.3%). There was a significant positive correlation between basal level of 2,3-DHBA and total reperfusion 2,3-DHBA content in SHRSP (r = 0.752; p < 0.05). Thus, data obtained clearly indicate that the hypertrophied myocardium of SHRSP was less tolerant to ischemia/reperfusion than that of Wistar rats due to chronically increased OH* production and enhanced total OH* output during reperfusion. Greater myocardial damage in SHRSP than in Wistar rats following the equal increase in OH* production above the basal level suggests the existence of deficit of the antioxidant defense in the hypertrophied myocardium.

Formation of hydroxyl radicals during myocardial reperfusion after experimental ischemia of different duration.

The intensity of hydroxyl radical (OH*) formation in the myocardium during reperfusion after ischemia of different duration was evaluated using microdialysis with sodium salicylate. 2,3-Dihydroxybenzoic acid, a product of OH* trapping by salicylic acid, was used as a marker of OH* generation in the postischemic myocardium. Experiments were performed on open-chest anesthetized and jet-ventilated Wistar rats. The concentrations of 2,3-dihydroxybenzoic acid in the dialysate were measured by high performance liquid chromatography (HPLC) with electrochemical detection. Experiments showed that the intensity and duration of free oxygen radical generation during reperfusion after 30-min ischemia far surpassed those observed after 20-min ischemia.

Perindopril effects on angiotensin I elimination in lung after experimental myocardial injury induced by intracoronary microembolization in rats.

The objective of the study was to determine whether angiotensin (Ang) I elimination in lung circulation depends on the degree of myocardial damage with and without early long-term perindopril treatment in a rat model of myocardial injury induced by intracoronary microembolization. Twenty-one days after surgery, steady-state arterial [125I]-Ang I and [125I]-Ang II blood concentrations were measured after high-performance liquid chromatography separation during i.v. infusion of [125I]-Ang I in three groups of male Wistar conscious rats: (a) sham-operated rats receiving saline (sham group, n = 6); (b) rats after coronary microembolization receiving saline (saline group, n = 7); and (c) rats after coronary microembolization receiving perindopril (2 mg/kg/day; from days 2-20 after embolization; perindopril group, n = 6). Ang I clearance and the Ang I-to-Ang II concentration ratio (R) were estimated. The embolization per se resulted in focal fibrosis, appearance of hypertrophic and dystrophic cardiac myocytes, and was accompanied by increased Ang I clearance (1,479 vs. 314 ml/min in sham group), 1.8-fold decreased [125I]-Ang II arterial level, and decreased R (0.5 vs. 1.2 in sham group; p < 0.05). Only Ang I concentrations and R were correlated with number of scars (r = -0.77; p < 0.05; and r = -0.82; p < 0.01, respectively). Captopril bolus (1 mg/kg, i.v.) caused similar reduction in [125I]-Ang II blood concentration in both sham and saline groups, but a significant increase of [125I]-Ang I blood concentration was detected in the sham group only. Thus in rats with coronary microembolization, a higher proportion of Ang I in lung circulation is eliminated by pathways independent of angiotensin-converting enzyme. In the perindopril group, a reduced number of scars (seven vs. 17 per slice in the saline group; p < 0.05), density of dystrophic and hypertrophic cardiac myocytes, and increased content of cell glycogen were observed. It was accompanied by normalized arterial [125I]-Ang I concentration, Ang I clearance, and R; [125I]-Ang II concentration tended to that in sham group. Only in the sham and perindopril groups was there significant correlation between Ang I and Ang II concentrations. The clear relation between number of scars per slice and R (r = -0.83; p < 0.01) was observed in all rats with embolized coronary vessels (saline and perindopril groups together). In conclusion, in this experimental, model Ang I elimination in the lung circulation was directly related to the degree of myocardial damage. Early perindopril treatment prevented maladaptive changes in Ang I processing and led to significant reduction of the undesirable aftereffects of myocardial tissue damage. Our data demonstrate the cardioprotective action of perindopril based on its beneficial influence on the renin-angiotensin system disturbances.

Systemic and regional hemodynamic effects of perindopril in experimental heart failure.

The effects of converting-enzyme inhibition by perindoprilat (0.5 mg/kg, intravenously, short-term administration) or perindopril (2 mg/kg, orally, long-term administration once a day for 21 days) on systemic and regional hemodynamics were studied on a new rat model of heart failure, which was induced by microembolization of coronary vessels by 15 microns plastic microspheres. Cardiac output and regional blood flows were measured by microsphere technique; the tone of the venous vessels was determined as mean circulatory filling pressure in conscious, freely moving rats. Perindoprilat evoked a much more prominent increase in kidneys, adrenal glands, intestine, and skin blood flows in embolized rats than in sham-operated rats. The differences between the effects of long-term treatment with perindopril in sham-operated and embolized rats were highly significant. Mean circulatory filling pressure was decreased by short-term and long-term administration of an angiotensin-converting enzyme inhibitor. It is concluded that venous vessels could be one of the target sites for the effects of perindopril-like drugs.

Simple method to produce acute heart failure by coronary vessel embolization in closed chest rats with microspheres.

Changes in left ventricle (LV) function, systemic, and regional hemodynamics as a result of coronary artery embolization by 15 microns microspheres were studied in rats. Selective coronary embolization was produced by injection of microspheres during ascending aorta occlusion animals by using an "L"-shaped wire in closed chest animals. Maximal developed LV systolic pressure (LVSPmax) was determined during ascending aorta occlusion. Coronary embolization evoked reductions in LVSPmax and +dP/dtmax and then decreased in basal LVSP, dP/dtmax, dP/dtmax/P, with a parallel increase in LV end-diastolic pressure (LVEDP). The number of microspheres accumulating in the heart following coronary embolization was about 40% of the total amount of the injected microspheres (300,000-400,000). In conscious rats 48 hr after coronary vessel embolization (in LV myocardium 100,003 +/- 4,334 microspheres per gram) the cardiac index, mean arterial pressure, -dP/dtmax and stroke volume were reduced by 35.6%, 20%, 17.2%, and 26.7%, respectively, when compared with sham-operated rats. LVEDP was increased by 40%, when compared with sham-operated rats. These results show that in this rat model of coronary vessel embolization heart failure develops. The model created may be used for the studies of pathophysiology of acute heart failure as well as for screening new compounds potentially effective in heart failure.

The cardiac contractile function and hemodynamic control in rats after chronic adriamycin treatment.

Cardiac contractile function and hemodynamic parameters of control and adriamycin-treated (2 mg/kg once a week for 10 weeks) rats were studied both in the anesthetized (hexenal, 20 mg/kg) and conscious state. Radiolabelled microspheres (diameter, 15 microns) were used to measure systemic and regional hemodynamics. No significant differences between the control and adriamycin-treated groups in cardiac contractile function, total peripheral resistance, and regional blood flow (except muscles) was found in anesthetized animals. In the conscious state, a significantly higher (+70%) total peripheral resistance combined with lower blood flow in the skin and spleen was observed in adriamycin-treated rats. The response of the heart rate to changes in the arterial pressure induced by nitroglycerin and phenylephrine injection was greatly diminished after adriamycin treatment. Isoprenaline (0.64 micrograms.kg-1.min-1) increased left ventricular contractile indices approximately twofold and heart rate by 30% in the control group, while in adriamycin-treated rats only minor changes in these parameters were observed. However, cardiac output rose by 36% and total peripheral resistance fell by 36% in these animals. Results show that prolonged adriamycin treatment leads to decreased inotropic response to beta-adrenoceptor stimulation and reduced baroreflex control. These changes occur in the stage preceding congestive heart failure.