Evaluation of cardiac troponin I and T levels as markers of myocardial damage in doxorubicin-induced cardiomyopathy rats, and their relationship with echocardiographic and histological findings.
BACKGROUND
Cardiac troponins I (cTnI) and T (cTnT) have been shown to be highly sensitive and specific markers of myocardial cell injury. We investigated the diagnostic value of cTnI and cTnT for the diagnosis of myocardial damage in a rat model of doxorubicin (DOX)-induced cardiomyopathy, and we examined the relationship between serial cTnI and cTnT with the development of cardiac disorders monitored by echocardiography and histological examinations in this model.


METHODS
Thirty-five Wistar rats were given 1.5 mg/kg DOX, i.v., weekly for up to 8 weeks for a total cumulative dose of 12 mg/kg BW. Ten rats received saline as a control group. cTnI was measured with Access(R) (ng/ml) and a research immunoassay (pg/ml), and compared with cTnT, CK-MB mass and CK. By using transthoracic echocardiography, anterior and posterior wall thickness, LV diameters and LV fractional shortening (FS) were measured in all rats before DOX or saline, and at weeks 6 and 9 after treatment in all surviving rats. Histology was performed in DOX-rats at 6 and 9 weeks after the last DOX dose and in all controls.


RESULTS
Eighteen of the DOX rats died prematurely of general toxicity during the 9-week period. End-diastolic (ED) and end-systolic (ES) LV diameters/BW significantly increased, whereas LV FS was decreased after 9 weeks in the DOX group (p<0.001). These parameters remained unchanged in controls. Histological evaluation of hearts from all rats given DOX revealed significant slight degrees of perivascular and interstitial fibrosis. In 7 of the 18 rats, degeneration and myocyte vacuolisation were found. Only five of the controls exhibited evidence of very slight perivascular fibrosis. A significant rise in cTnT was found in DOX rats after cumulative doses of 7.5 and 12 mg/kg in comparison with baseline (p<0.05). cTnT found in rats after 12 mg/kg were significantly greater than that found after 7.5 mg/kg DOX. Maximal cTnI (pg/ml) and cTnT levels were significantly increased in DOX rats compared with controls (p=0.006, 0.007). cTnI (ng/ml), CK-MB mass and CK remained unchanged in DOX rats compared with controls. All markers remained stable in controls. Analysis of data revealed a significant correlation between maximal cTnT and ED and ES LV diameters/BW (r=0.81 and 0.65; p<0.0001). A significant relationship was observed between maximal cTnT and the extent of myocardial morphological changes, and between LV diameters/BW and histological findings.


CONCLUSIONS
Among markers of ischemic injury after DOX in rats, cTnT showed the greatest ability to detect myocardial damage assessed by echocardiographic detection and histological changes. Although there was a discrepancy between the amount of cTnI and cTnT after DOX, probably due to heterogeneity in cross-reactivities of mAbs to various cTnI and cTnT forms, it is likely that cTnT in rats after DOX indicates cell damage determined by the magnitude of injury induced and that cTnT should be a useful marker for the prediction of experimentally induced cardiotoxicity and possibly for cardioprotective experiments.
Bertinchant
J P
JP
Laboratory of Cardiovascular Physiology, University of Montpellier-Nîmes, France. jean.pierre.bertinchant@chu-nimes.fr
Polge
A
A
Juan
J M
JM
Oliva-Lauraire
M C
MC
Giuliani
I
I
Marty-Double
C
C
Burdy
J Y
JY
Fabbro-Peray
P
P
Laprade
M
M
Bali
J P
JP
Granier
C
C
de la Coussaye
J E
JE
Dauzat
M
M
D015153:Blotting, Western; D002401:Cathepsin B / Q000134:cerebrospinal fluid*; D056655:Cathepsin H; D002403:Cathepsins / Q000134:cerebrospinal fluid*; D002452:Cell Count; D002555:Cerebrospinal Fluid / Q000166:cytology; D055316:Cystatin C; D015891:Cystatins / Q000134:cerebrospinal fluid*; D003546:Cysteine Endopeptidases / Q000134:cerebrospinal fluid*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D007938:Leukemia / Q000473:pathology; D008577:Meningeal Neoplasms / Q000134:cerebrospinal fluid* / Q000473:pathology / Q000556:secondary*; D009362:Neoplasm Metastasis
C525030:CST3 protein, human; C525032:Cst3 protein, rat; D055316:Cystatin C; D015891:Cystatins; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; C534126:CTSH protein, human; D056655:Cathepsin H; C534128:Ctsh protein, rat