Direct Effects of Toxic Divalent Cations on Contractile Proteins with Implications for the Heart: Unraveling Mechanisms of Dysfunction.

The binding of calcium and magnesium ions to proteins is crucial for regulating heart contraction. However, other divalent cations, including xenobiotics, can accumulate in the myocardium and enter cardiomyocytes, where they can bind to proteins. In this article, we summarized the impact of these cations on myosin ATPase activity and EF-hand proteins, with special attention given to toxic cations. Optimal binding to EF-hand proteins occurs at an ionic radius close to that of Mg2+ and Ca2+. In skeletal Troponin C, Cd2+, Sr2+, Pb2+, Mn2+, Co2+, Ni2+, Ba2+, Mg2+, Zn2+, and trivalent lanthanides can substitute for Ca2+. As myosin ATPase is not a specific MgATPase, Ca2+, Fe2+, Mn2+, Ni2+, and Sr2+ could support myosin ATPase activity. On the other hand, Zn2+ and Cu2 significantly inhibit ATPase activity. The affinity to various divalent cations depends on certain proteins or their isoforms and can alter with amino acid substitution and post-translational modification. Cardiac EF-hand proteins and the myosin ATP-binding pocket are potential molecular targets for toxic cations, which could significantly alter the mechanical characteristics of the heart muscle at the molecular level.

Characteristics of the right atrial and right ventricular contractility in a model of monocrotaline-induced pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) leads to changes in the pump function of the heart and causes right-sided myocardial hypertrophy and heart failure. This study was the first to compare the contractile characteristics of the multicellular myocardial preparations of the right atrium (RA) and right ventricle (RV) of male rats from the control group (CON) and the group with monocrotaline (MCT)-induced hypertrophy at the molecular and multicellular levels. In both RA and RV in MCT-treated rats, the fraction of motile filaments and the maximum sliding velocity of actin and reconstituted thin filaments over myosin decreased, and the ratio of α-/ß-myosin heavy chains (MHC) shifted towards ß-MHC. In the RA strips and RV trabeculae, the maximum shortening velocity, the extent of muscle shortening, the amplitude of isometric stress, the amount of work decreased. PAH leads to a greater drop in right atrial contractility than that of the ventricle.

Molecular mechanisms of mechanical function changes of the rat myocardium under subchronic lead exposure.

A moderate degree of lead intoxication was observed in male rats after repeated intraperitoneal injections with two doses of lead acetate three times a week during 5 (12.5 mg of Pb per kg body mass) and 6 (6.01 mg of Pb per kg body mass) weeks. Using an in vitro motility assay, we investigated the impact of this intoxication on the characteristics of actin-myosin interaction and its regulation in the atria, right, and left ventricles. Both lead doses exposure decreased the maximum sliding velocity of reconstituted thin filaments over myosin and fraction of motile filaments in all heart chambers, caused the myosin isoforms shift towards slower ß-myosin heavy chains in ventricles and decreased regulatory light chain phosphorylation in atria. No statistically significant difference was found in force and calcium regulation of actin-myosin interaction. A dose-dependent effect of lead on myosin functional characteristics was found in all heart chambers, but the degree of this effect varied depending on the heart chamber.

Changes in the Cardiotoxic Effects of Lead Intoxication in Rats Induced by Muscular Exercise.

Exposure to lead is associated with an increased risk of cardiovascular diseases. Outbred white male rats were injected with lead acetate intraperitoneally three times a week and/or were forced to run at a speed of 25 m/min for 10 min 5 days a week. We performed noninvasive recording of arterial pressure, electrocardiogram and breathing parameters, and assessed some biochemical characteristics. Electrophoresis in polyacrylamide gel was used to determine the ratio of myosin heavy chains. An in vitro motility assay was employed to measure the sliding velocity of regulated thin filaments on myosin. Isolated multicellular preparations of the right ventricle myocardium were used to study contractility in isometric and physiological modes of contraction. Exercise under lead intoxication normalized the level of calcium and activity of the angiotensin-converting enzyme in the blood serum, normalized the isoelectric line voltage and T-wave amplitude on the electrocardiogram, increased the level of creatine kinase-MB and reduced the inspiratory rate. Additionally, the maximum sliding velocity and the myosin heavy chain ratio were partly normalized. The effect of exercise under lead intoxication on myocardial contractility was found to be variable. In toto, muscular loading was found to attenuate the effects of lead intoxication, as judged by the indicators of the cardiovascular system.

Cardioinotropic Effects in Subchronic Intoxication of Rats with Lead and/or Cadmium Oxide Nanoparticles.

Subchronic intoxication was induced in outbred male rats by repeated intraperitoneal injections with lead oxide (PbO) and/or cadmium oxide (CdO) nanoparticles (NPs) 3 times a week during 6 weeks for the purpose of examining its effects on the contractile characteristics of isolated right ventricle trabeculae and papillary muscles in isometric and afterload contractions. Isolated and combined intoxication with these NPs was observed to reduce the mechanical work produced by both types of myocardial preparation. Using the in vitro motility assay, we showed that the sliding velocity of regulated thin filaments drops under both isolated and combined intoxication with CdO-NP and PbO-NP. These results correlate with a shift in the expression of myosin heavy chain (MHC) isoforms towards slowly cycling ß-MHC. The type of CdO-NP + PbO-NP combined cardiotoxicity depends on the effect of the toxic impact, the extent of this effect, the ratio of toxicant doses, and the degree of stretching of cardiomyocytes and muscle type studied. Some indices of combined Pb-NP and CdO-NP cardiotoxicity and general toxicity (genotoxicity included) became fully or partly normalized if intoxication developed against background administration of a bioprotective complex.

Force-velocity characteristics of isolated myocardium preparations from rats exposed to subchronic intoxication with lead and cadmium acting separately or in combination.

This investigation continues our study of the effects of Pb-Cd poisoning on the heart, extending the enquiry from isometric to auxotonic contractions, thereby examining the effect on the ability of myocardial tissues to perform mechanical work. Different shifts were revealed in myocardial force-velocity relations following subchronic exposure of rats to lead acetate and cadmium chloride acting separately, in combination, or in combination with a bioprotective complex (BPC). The experiments were conducted on isolated preparations of trabecules and papillary muscles of the right ventricle in physiological loading conditions and on isolated heart muscle contractile proteins examined by the in vitro motility assay. The results of the latter correlate with the shifts in the ratio of cardiac myosin isoforms. The amount of work performed by the myocardium was calculated on the basis of the tension-shortening loop area and was found to be similar in the preparations from all experimental groups. This fact presumably reflects adaptive capacity of the myocardial function even when contractility is damaged due to the metallic intoxication of a moderate severity. Some characteristics of rat myocardium altered by the impact of lead-cadmium intoxication became fully or partly normalized if intoxication developed against background administration of a bioprotective complex (BPC). Together with previously reported results obtained in the isometric mode of contractility, all these results strengthen the scientific foundations of risk assessment and risk management projects in the occupational and environmental conditions characterized by human exposure to lead and/or cadmium.

Changes in rat myocardium contractility under subchronic intoxication with lead and cadmium salts administered alone or in combination.

Subchronic intoxications induced in male rats by repeated intraperitoneal injections of lead acetate and cadmium chloride, administered either alone or in combination, are shown to affect the biochemical, cytological and morphometric parameters of blood, liver, heart and kidneys. The single twitch parameters of myocardial trabecular and papillary muscle preparations were measured in the isometric regime to identify changes in the heterometric (length-force) and chronoinotropic (frequency-force) contractility regulation systems. Differences in the responses of these systems in trabecules and papillary muscles to the above intoxications are shown. A number of myocardium mechanical characteristics changing in rats under the effect of a combined lead-cadmium intoxication and increased proportion of α-myosin heavy chains were observed to normalize fully or partially if such intoxication was induced against background administration of a proposed bioprotective complex. Based on the experimental results and literature data, some assumptions are suggested concerning the mechanisms of the cardiotoxic effects produced by lead and cadmium.

Further analysis of rat myocardium contractility changes associated with a subchronic lead intoxication.

A moderate subchronic lead intoxication was observed in male rats after repeated intraperitoneal injections of lead acetate. Right ventricular trabeculae and papillary muscles were isolated for in vitro studying of the contraction-relaxation cycle under isotonic and physiological loading. The contractile function of the myocardium was also assessed by measuring the velocity of thin filament movement over myosin. Lead intoxication led in papillary muscles to a decrease in the maximal rate of isotonic shortening for all afterloads and a decrease in the thin filament sliding velocity. Papillary muscles from lead-exposed rats displayed marked changes in most of the main characteristics of afterload contraction-relaxation cycles, but in trabeculae these changes were less pronounced. The reported changes were attenuated to some extent in rats treated with a Ca-containing bioprotector. The amount of work produced by both types of heart muscle preparations was not changed by lead. Only in papillary muscles the load-dependent relaxation index was significantly increased in the lead-treated groups. Thus subchronic lead intoxication affects the peak rate of force development and relaxation properties of cardiac muscle contracting in isotonic/physiological regimes rather than the total amount of mechanical work, which may reflect adaptive changes in the myocardial function under decreased contractility.

Effects of subchronic lead intoxication of rats on the myocardium contractility.

Outbred male rats were repeatedly injected IP with sub-lethal doses of lead acetate 3 times a week during 5 weeks. They developed an explicit, even if moderate, lead intoxication characterized by typical hematological and some other features. The next day after the last injection the heart of each animal was excised, and the trabecules and papillary muscles from the right ventricle were used for modeling in vitro isometric (with varying starting length of the preparation) regimes of the contraction-relaxation cycle with different preloads. Several well-established parameters of this model were found changed compared with the preparations taken from the hearts of healthy control rats. Background in vivo calcium treatment attenuated both systemic and cardiotoxic effects of lead to an extent. We show for the first time that subchronic intoxication with lead caused myocardial preparations in a wide range of lengths to respond by a decrease in the time and speed parameters of the isometric contraction while maintaining its amplitude and by a decrease in the passive stiffness of trabecules. The responses of the various heart structures are outlined, and the isomyosin ratio is shown to have shifted towards the slow isoform. Mechanistic and toxicological inferences from the results are discussed.