Serum Creatine Phosphokinase Isoenzyme Research Articles

Sesamol Attenuates Isoproterenol-induced Acute Myocardial Infarction via Inhibition of Matrix Metalloproteinase-2 and -9 Expression in Rats

Background/Aims: The protective role of sesamol and its possible action against isoproterenol-induced myocardial injury and infarction is unknown. We tested the hypothesis that sesamol’s protection against myocardial infarction is associated with the inhibition of matrix metalloproteinase (MMP)-2 and MMP-9. Methods: Four groups of experimental rats were subcutaneously injected with sesamol (0, 1, 3, or 10 mg/kg) and then, 2 h later, intraperitoneally injected with isoproterenol (100 mg/kg 24 h apart on 2 consecutive days) to induce myocardial infarction. Control rats were treated with saline only. Blood pressure (BP), heart rate (HR), and electrocardiography (ECG) wave durations, serum creatine phosphokinase isoenzymes (CKMB), lactate dehydrogenase (LDH), myocardial histology, MMP-2, and MMP-9 were assessed 24 h after the last dose of isoproterenol was given. Results: BP was lower, and HR, ECG wave durations, CKMB, LDH, myocardial injury, MMP-2, and MMP-9 levels were higher in experimental rats than in control rats. BP was significantly higher, and all the other parameters were significantly lower in the rats treated with sesamol than in those treated with isoproterenol only. Conclusions: Sesamol effectively prevented myocardial infarction, at least in part, by controlling proteolytic activities and the expression of MMP-2 and -9 in isoproterenol-treated rats.

Carnitine Deficiency Aggravates Cyclophosphamide-Induced Cardiotoxicity in Rats

Background: This study examined, for the first time, the involvement of carnitine deficiency in cardiotoxicity, particularly cyclophosphamide (CP)-induced cardiomyopathy, as well as effects of carnitine supplementation with propionyl-L-carnitine (PLC) on cardiotoxicity. Methods: An animal model of carnitine deficiency was developed in rats treated with D-carnitine (DC)-mildronate (MD). Adult male Wistar albino rats were assigned to one of six treatment groups: the first three groups were injected intraperitoneally with normal saline, PLC (250 mg/kg/day), and DC (250 mg/kg/day) combined with MD (200 mg/kg/day), respectively, for 10 successive days. In groups 4–6, the same doses of normal saline, PLC and DC-MD were injected, respectively, during the 5 successive days before and after a single dose of CP (200 mg/kg). On day 6 after CP treatment, 24-hour urine was collected, then animals were sacrificed, and serum as well as hearts were isolated. Results: CP caused a significant increase in serum creatine phosphokinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), urinary carnitine excretion and clearance and intramitochondrial acetyl-CoA/CoA-SH, and a significant decrease in serum free carnitine, total carnitine and adenosine triphosphate (ATP) contents in cardiac tissue. In the carnitine-depleted rats, CP induced dramatic increases in CK-MB and LDH levels, carnitine clearance and intramitochondrial acetyl-CoA/CoA-SH, as well as progressive reduction in total carnitine and ATP in cardiac tissues. Interestingly, PLC supplementation completely reversed the biochemical and histopathological changes induced by CP to the control values. Conclusion: (1) Carnitine deficiency is a risk factor which is involved in CP-related cardiomyopathy; (2) serum and urinary carnitine levels should be monitored and viewed as indices of CP-induced multiple organ toxicity, and (3) carnitine supplementation, using PLC, prevents the development of CP-induced cardiotoxicity.

Protection of myocardiocytes against anoxia-reoxygeneration injury by heat shock protein 70 gene transfection: experiment with rats

To study the protective role of heat shock protein (HSP)70 expression in the myocardial cells undergoing anoxia/reoxygeneration. Myocardiocytes of neonatal SD rats were cultured and randomly divided into 4 groups: anoxia/reperfusion (A/R) group undergoing anoxia/reperfusion; A/R + HS group, undergoing transfection of HS and A/R, pCDNA HSP70 + A/R group, undergoing transfection of liposome-coated HSP70 pCDNA and A/R, and control group. RT-PCR and Western blotting were used to detect the mRNA and protein expression of HSP70 in the cells. The cell viability was examined by MTT method. The levels of lactic dehydrogenase (LDH), and serum creatine phosphokinase isoenzyme (CPK) were detected. Transmission electronic microscopy was used to observe the ultrastructure of the cells. Apoptosis rate was measured. The cell viability rates of the HS + A/R group and pCDNA HSP70 + A/R group were (71.8 +/- 10.3)% and (73.4 +/- 12.2)% respectively, both significantly higher than that of the A/R group [(35.2 +/- 6.8)%, both P < 0.01]. The LDH activity levels of the HS + A/R and pCDNA HSP70 + A/R groups were (14.3 +/- 2.6) and (14. 6 +/- 2.9) IU/L respectively, both significantly lower than that of the A/R group [22.9 +/- 4.0 IU/L, P < 0.01]. The cell ultrastructure was abnormal in the A/R group e, whereas nearly normal in the HS + A/R and pCDNA HSP70 + A/R groups. The HSP70 mRNA land protein were only slightly expressed in the myocardiocytes of the A/R group; However, the expressions of HSP70 mRNA and protein were of the HS + A/R and pCDNA HSP70 + A/R groups were significantly higher than those of the A/R group (both P < 0.01). The apoptosis rates of the A/R group was (12.84 +/- 2.17), significantly higher than that of the control group [(2.16 +/- 0.15), P < 0.01], and the apoptotic rates of the HS + A/R and pCDNA HSP70 + A/R groups were (5.23 +/- 1.04) and (4.42 +/- 0.93) respectively, both significantly lower than that of the A/R group (P < 0.01). Overexpression of HSP70 alone can provide protection, related to the anti-apoptosis effect, for cardiomyocytes against anoxia-reoxygeneration.

Propionyl- l-carnitine prevents the progression of cisplatin-induced cardiomyopathy in a carnitine-depleted rat model

This study has been initiated to investigate whether endogenous carnitine depletion and/or carnitine deficiency is a risk factor during development of cisplatin (CDDP)-induced cardiomyopathy and if so, whether carnitine supplementation by propionyl- l-carnitine (PLC) could offer protection against this toxicity. To achieve the ultimate goal of this study, a total of 60 adult male Wistar albino rats were divided into six groups. The first three groups were injected intraperitoneally with normal saline, PLC (500 mg kg −1), and d-carnitine (500 mg kg −1) respectively, for 10 successive days. The 4th, 5th, and 6th groups were injected intraperitoneally with the same doses of normal saline, PLC and d-carnitine, respectively, for 5 successive days before and after a single dose of CDDP (7 mg kg −1). On day 6 after CDDP treatment, animals were sacrificed, serum as well as hearts were isolated and analyzed. CDDP resulted in a significant increase in serum creatine phosphokinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH), thiobarbituric acid reactive substances (TBARS) and total nitrate/nitrite (NO( x)) and a significant decrease in reduced glutathione (GSH), total carnitine, and adenosine triphosphate (ATP) content in cardiac tissues. In the carnitine-depleted rat model, CDDP induced dramatic increase in serum cardiomyopathy enzymatic indices, CK-MB and LDH, as well as progressive reduction in total carnitine and ATP content in cardiac tissue. Interestingly, PLC supplementation resulted in a complete reversal of the increase in cardiac enzymes, TBARS and NO x , and the decrease in total carnitine, GSH and ATP, induced by CDDP, to the control values. Moreover, histopathological examination of cardiac tissues confirmed the biochemical data, where PLC prevents CDDP-induced cardiac degenerative changes while d-carnitine aggravated CDDP-induced cardiac tissue damage. In conclusion, data from this study suggest for the first time that carnitine deficiency and oxidative stress are risk factors and should be viewed as mechanisms during development of CDDP-related cardiomyopathy and that carnitine supplementation, using PLC, prevents the progression of CDDP-induced cardiotoxicity.

Cardiotoxicity of doxorubicin/paclitaxel combination in rats: effect of sequence and timing of administration.

The higher incidence of cardiotoxicity of doxorubicin (DOX)/paclitaxel (PTX) combination compared with DOX alone remains to be a major obstacle against effective chemotherapeutic treatment. We investigated the effect of sequence and time interval between administration of both drugs on the severity of cardiotoxicity of the combination. Male Wistar rats were divided into seven groups. DOX was administered intraperitoneally (i.p.) at a single dose of 5 mg x kg(-1) every other 2 days, 2 doses per week for a total cumulative dose of 20 mg x kg(-1). PTX was administered by an i.p. route at a dose of 20 mg x kg(-1) every other 2 days. Both drugs were injected either alone or sequentially in combination. In one case, DOX preceded PTX by 30 min and 24 h and in the other case, PTX preceded DOX by 30 min and 24 h. Cardiotoxicity was evaluated by both biochemical and histopathological examination, 48 h after the last DOX dose. DOX-induced cardiotoxicity was manifested by abnormal biochemical changes including marked increases in serum creatine phosphokinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), glutathione peroxidase (GSH-Px), and aspartate aminotransferase (AST) activity levels. Myocardial tissue from DOX-treated rats showed significant increases in malondialdehyde (MDA) production and total nitrate/nitrite (NOx) levels, parallel with depletion of "endogenous antioxidant reserve," including GSH contents and GSH-Px activity level. PTX treatment produced significant changes in the biochemical parameters measured by a lower magnitude than those changes produced by DOX alone. Combination of both drugs resulted in aggravation of DOX-induced cardiotoxicity regardless the sequence and time interval between administration of either drug. Administration of PTX 30 min and 24 h after DOX treatment showed exaggeration of combination-induced cardiotoxicity compared with the reverse sequence. This exacerbation was manifested by much more pronounced changes in serum and cardiac tissue parameters measured. Histopathological examination of ventricles of rat's heart revealed that DOX treatment produced myo-cytolysis and myocardial necrosis. Administration of PTX following DOX treatment showed extensive myocardial necrosis compared with those rats treated with either DOX alone or the reverse sequence of administration. Moreover, rats treated with PTX 24 h after DOX treatment showed exaggeration of the combination-induced cardiotoxicity. In conclusion, PTX might synergistically aggravate DOX-induced cardiotoxicity. The effect might be much more pronounced with those rats treated with PTX 24 h after DOX treatment.

Probucol Attenuates Cyclophosphamide‐induced Oxidative Apoptosis, p53 and Bax Signal Expression in Rat Cardiac Tissues

Cyclophosphamide (CP) is a widely used drug in cancer chemotherapy and immunosuppression, which could cause toxicity of the normal cells due to its toxic metabolites. Probucol, a cholesterol-lowering drug, acts as potential inhibitor of DNA damage and shows to protect against doxorubicin-induced cardiomyopathy by enhancing the endogenous antioxidant system including glutathione peroxidase, catalase and superoxide dismutase. This study examined the possible protective effects of probucol, a lipid-lowering compound with strong antioxidant properties, against CPinduced cardiotoxicity. This objective could be achieved through studying the gene expression-based on the possible protective effects of probucol against CP-induced cardiac failure in rats. Adult male Wistar albino rats were assigned into four treatment groups: Animals in the first (control) and second (probucol) groups were injected intraperitoneally with corn oil and probucol (61 mg/kg/day), respectively, for two weeks. Animals in the third (CP) and fourth (probucol plus CP) groups were injected with the same doses of corn oil and probucol (61 mg/kg/day), respectively, for one week before and one week after a single dose of CP (200 mg/kg, I.P.). The p53, Bax, Bcl2 and oxidative genes signal expression were measured by real time PCR. CP-induced cardiotoxicity was clearly observed by a significant increase in serum creatine phosphokinase isoenzyme (CK-MB) (117%), lactate dehydrogenase (LDH) (64%), free (69%) and esterified cholesterol (42%) and triglyceride (69%) compared to control group. In cardiac tissues, CP significantly increases the mRNA expression levels of apoptotic genes, p53 with two-fold and Bax with 1.6-fold, and decreases the anti-apoptotic gene Bcl2 with 0.5-fold. Moreover, CP caused downregulation of antioxidant genes, glutathione peroxidase, catalase, and superoxide dismutase and increased the lipid peroxidation and decreased adenosine triphosphate (ATP) (40%) and ATP/ADP (44%) in cardiac tissues. Probucol pretreatment not only counteracted significantly the CP-induced increase in cardiac enzymes and apoptosis but also induced a significant increase in mRNA expression of antioxidant enzymes and improved ATP, ATP/ADP, glutathione (GSH) in cardiac tissues. In conclusion, data from the present study suggest that probucol prevents the development of CP-induced cardiotoxicity by a mechanism related, at least in part, to its ability to increase mRNA expression of antioxidant genes and to decrease apoptosis in cardiac tissues with the consequent improvement in mitochondrial oxidative phosphorylation and energy production.

Serum Enzyme Monitoring in Asthma Patients

To the Editor.— Maguire et al1 have recommended serial monitoring of serum creatine phosphokinase isoenzyme (CPK-MB) levels and electrocardiograms (ECG) for all children requiring admission to an intensive care unit for treatment of severe asthma. I wish to point out what I believe are inaccurate observations and invalid conclusions. In this study 20 admissions to intensive care units occurred for "severe bronchial obstruction despite standard initial therapy." Six necessitated use of intravenous isoproterenol for Pco2, values of 50 mm Hg or higher.

Cardiotoxicity During Treatment of Severe Childhood Asthma

We prospectively evaluated 20 patient admissions for severe exacerbation of childhood asthma at The Children's Hospital, Boston, to detect evidence of cardiotoxicity. Evidence of cardiotoxicity was found in all six patient admissions for which isoproterenol infusion was utilized. This included marked elevation of serum creatine phosphokinase isoenzyme (CPK-MB) levels and electrocardiogram abnormalities consistent with transient myocardial ischemia. Peak serum CPK-MB levels were significantly lower and electrocardiogram abnormalities were significantly less frequent during 14 patient admissions for which isoproterenol infusion was not utilized. Risk factors associated with cardiotoxicity included tachycardia, hypercapnia, acidosis, and intravenous isoproterenol therapy. We conclude that cardiotoxicity is not infrequent during therapy for severe exacerbations of childhood asthma. Electrocardiograms and measurement of serum CPK-MB levels are sensitive, useful, and readily obtained indicators of cardiotoxicity. Abnormalities of these studies may detect cardiotoxicity prior to the occurrence of more blatant or catastrophic manifestations of cardiotoxicity. We therefore recommend serial monitoring of serum CPK-MB levels and electrocardiograms for all children requiring an admission to the intensive care unit for management of severe asthmatic exacerbation.

Transient electrocardiographic changes suggesting myocardial ischaemia in newborn infants following tocolysis with beta-sympathomimetics

Serial electrocardiograms (ECGs) were studied prospectively in 80 apparently healthy newborn infants; 30 infants exposed in utero to prolonged tocolytic therapy (21 to ritodrine and 9 to isoxsuprine) and 50 infants non-exposed in utero to drugs (control group) matched for gestational age, Apgar score, and birth weight. Duration of exposure to tocolysis was at least 30 days (30-180 days) with an oral dosage of 10 mg 3 times daily. ECGs were graded for changes suggestive of ischaemia using the arbitrary grading system described by Jedeikin et al. In all infants with ECG features of myocardial ischaemia, serum creatine-phosphokinase iso-enzyme (CK-MB) activity was measured. Six out of 21 infants to ritodrine and six out of nine infants exposed to isoxsuprine showed a degree of ECG ischaemia which persisted for several weeks. No control infant presented grade 2 or 3 ECG changes after the 5th day of life. The results of this study seem to show that prolonged tocolytic therapy with beta-sympathomimetics has side-effects on the fetal myocardium and suggest that this treatment be reserved only for selective cases and/or for short periods of time.

Sequential Changes in the Serum Creatine Phosphokinase Isoenzyme Activity and Correlation with Prognosis in Patients with Acute Head Injuries

A sensitive biochemical marker for acute brain damage is important in assessing the severity of head injuries. Acute brain damage is followed by increased creatine phosphokinase (CPK) activity in the serum. The present study was undertaken to examine whether the measurement of serum levels of brain type isoenzyme (CPK-BB) and heart type isoenzyme (CPK-MB) is valuable in assessing and evaluating the clinical status of patients with acute head injuries. Fifty-seven patients with acute head injuries were studied. Fourty-four patients were male and thirteen patients were female with ages ranging from 2 to 69 years. All patients were diagnosed by computed tomography (CT) scan within three hours after head injury. Serial blood samples were drawn during the first 7 days after head injury. The Glasgow Coma Scale (GCS) for all patients was recorded on admission. Clinical outcome was assessed at the time of discharge according to the Glasgow Outcome Scale. The “good” group consisted of patients with good recovery or moderate disability. The “poor” group consisted of patients with severe disability and persistent vegetative state. The maximal serum levels of CPK-BB and CPK-MB occurred in most patients on the first day of trauma. There was a highly significant correlation between the serum levels of CPK-BB and GCS (p<0.001), whereas there was little correlation between serum levels of CPK-MB and GCS. The peak levels of CPK-BB in the “poor” and “dead” groups were significantly higher than those in the “good” group (p<0.01). Particular correlation between CT findings and serum levels of CPK-BB was found. Cases with serum levels of CPK-BB higher than 10 IU/l were associated with cerebral contusions. The major factors in the increase of serum levels of CPK-BB were associated with the severity of cerebral contusions and particular sites of the brain injuries. New lesions in serial CT scans didn't seem to be a factor in raising serum levels of CPK-BB. Serum levels of CPK-BB did not correlate with intracranial pressure. Measurement of serum levels of CPK-BB may offer useful information for making a prognosis. It seems to be a sensitive index of acute brain damage and a reliable marker of cerebral contusions.

Serum CPK isoenzyme BB as an indicator of brain tissue damage following head injury.

In a study of 209 patients who had serum drawn for assay of creatine phosphokinase (CPK) isoenzymes, an attempt was made to correlate elevation of CPK brain fraction (BB) with occurrence of brain injury and to evaluate its sensitivity as an indicator of the presence of brain damage. Thirty-three patients had suffered serious head trauma. However, of the ten patients in coma, only three had elevated serum CPK-BB levels. Twenty-four of the other 176 patients who had no history and head injury also had positive BB fractions. Serum CPK-BB level was therefore an unreliable indicator of the extent of brain damage.

Specificity of elevated serum MB creatine phosphokinase activity in the diagnosis of acute myocardial infarction

Creatine phosphokinase (CPK) isoenzyme determinations are useful in the diagnosis of myocardial infarction. However, until suitably sensitive and precise quantitative procedures became available, the diagnostic specificity of serum CPK isoenzyme elevations could not be thoroughly examined. In this study an assay procedure capable of accurately determining activity of individual CPK isoenzymes even in serum samples with normal total CPK activity was employed to obtain two types of information. First, CPK isoenzyme profiles were examined in extracts of a spectrum of human tissues obtained at operation to determine whether the isoenzyme associated with myocardium is present in other human tissues in quantities sufficient to produce increased activity in serum. In addition, CPK isoenzymes were analyzed quantitatively in serial serum samples from 50 hospitalized control subjects, 100 patients with acute myocardial infarction, 100 patients undergoing noncardiac surgery and 50 patients undergoing cardiac catheterization to determine whether insult to tissues other than the heart is associated with increased “myocardial” CPK isoenzyme activity in serum. Results from analyses of tissue extracts indicated that myocardium is the only tissue surveyed containing sufficient MB CPK to account for substantial increases in serum MB activity. Results from analyses of serial serum samples indicated that MB CPK activity levels are consistently elevated after myocardial infarction, averaging 0.089 IU/ml. However, after cardiac catheterization or noncardiac surgery peak serum MB activity remains low, averaging only 0.004 IU/ml despite marked elevations in total serum CPK activity. Thus, elevated serum MB CPK activity is a highly specific as well as sensitive criterion of myocardial injury.

Serum Creatine Phosphokinase Isoenzyme (CPK2) in Myositis

Serum creatine phosphokinase (CPK) isoenzyme levels were studied in six cases: two with polymyositis, two with dermatomyositis, and two with viral myositis without neurological involvement. In all patients, agarose gel fractionation and fluorometric quantitation of serum CPK isoenzymes showed the presence of CPK₂, although there was no evidence of myocardial infarction. Thus, CPK₂is not exclusively found in myocardial infarction. The prolonged elevation of CPK₂levels during periods of acute exacerbation of myositis in the reported cases is in contrast to the usual transient increases in myocardial infarction. Levels of serum lactate dehydrogenase (LDH), aspartate aminotransferase, and aldolase were invariably increased in these patients. The expected LDH₅increase with myositis was not found. (<i>JAMA</i>230:1141-1144, 1974)

Serum creatine phosphokinase isoenzyme (CPK2) in myositis. A report of six cases

Serum creatine phosphokinase isoenzyme (CPK2) in myositis. A report of six cases

Serum Creatine Phosphokinase and Myocardial Infarction

ELEVATED level of serum creatine phosphokinase (CPK) activity is a well recognized manifestation of acute myocardial infarction. Recently, total CPK and CPK isoenzyme determinations have been utilized to improvedetection, quantification, andprojectionof the evolution of infarction as well. Improved Detection Three CPK isoenzymes (MM, BB, MB) may appear in serum—each a dimer composed of two subunits of the M or B type. Myocardium contains the MB form as well as MM in contrast to skeletal muscle. Myocardial injury is associated with elevation of MB CPK levels. In contrast, after injury to skeletal muscle, serum CPK increases reflect MM activity. By detecting electrophoretically separated serum CPK isoenzymes with a fluorescence assay affording increased sensitivity, Roe and his colleagues have substantially facilitated the detection of myocardial infarction. Although they have cautioned that elevated levels of MB activity are not uniquely associated with myocardial injury, they have demonstrated that isoenzyme MB

Serum creatine phosphokinase and myocardial infarction

ELEVATED level of serum creatine phosphokinase (CPK) activity is a well recognized manifestation of acute myocardial infarction. Recently, total CPK and CPK isoenzyme determinations have been utilized to improve<i>detection, quantification</i>, and<i>projection</i>of the evolution of infarction as well. <h3>Improved Detection</h3> Three CPK isoenzymes (MM, BB, MB) may appear in serum—each a dimer composed of two subunits of the M or B type. Myocardium contains the MB form as well as MM in contrast to skeletal muscle. Myocardial injury is associated with elevation of MB CPK levels. In contrast, after injury to skeletal muscle, serum CPK increases reflect MM activity. By detecting electrophoretically separated serum CPK isoenzymes with a fluorescence assay affording increased sensitivity, Roe and his colleagues have substantially facilitated the detection of myocardial infarction. Although they have cautioned that elevated levels of MB activity are not uniquely associated with myocardial injury, they have demonstrated that isoenzyme MB

Quantification of serum creatine phosphokinase isoenzyme activity

Three serum creatine phosphokinase (CPK) isoenzymes (MM, MB and BB) have been recognized. Myocardium is richly endowed with MB CPK. Accordingly, increased serum MB CPK activity reflects myocardial injury. Although recent advances have facilitated detection of activity of CPK isoenzymes, available techniques have unavoidable quantitative limitations. Accordingly, we used a new procedure to quantify CPK isoenzymes in serum samples with normal or increased total CPK activity. Samples were diluted or concentrated so that total CPK activity was 0.100 to 1.200 IU/ml and then subjected to electrophoresis on cellulose acetate. Regions of the strips encompassing each isoenzyme were cut out, immersed in 0.5 to 3 ml of assay medium in which the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) was generated as a function of isoenzyme activity. NADPH was detected by serial determinations of fluorescence in the medium. The assay was linear with respect to time and isoenzyme activity (0.010 to 1.2 ID/ml, no. = 100) and recovery averaged 83 ± 1.8 percent (standard deviation). Reproducibility was within 2 percent. Accuracy of the method was validated with selected mixtures of BB and MM isoenzymes. Serum MB CPK activity averaged only 0.002 ± 0.001 lU/ml in normal persons ( no. = 27), compared to 0.064 ± 0.043 lU/ml in 30 patients with myocardial infarction. Serum MB CPK activity was not increased in 23 patients after intramuscular injections or abdominal surgery. The procedure described permits quantitative determination of activity of individual CPK isoenzymes in serum samples with or without increased total CPK activity and thus is of potential value in the quantitative assessment of myocardial infarction by serum enzyme analysis.

Serum creatine phosphokinase isoenzymes in congenital hypothyroidism

Summary By means of electrophoresis the serum CPK isoenzyme pattern was examined in patients with congenital hypothyroidism. Only the muscle isoenzyme was demonstrated on the electrophoretic pattern. This observation suggests that serum CPK in congenital hypothyroidism derives from muscle fibers.