Title: Effect of carbon monoxide releasing molecule-3 on oxygen-glucose deprivation and restoration injury to rat cardiomyocytes
Abstract: Objective
To evaluate the effect of carbon monoxide-releasing molecule-3(CORM-3)on oxygen-glucose deprivation and restoration(OGD/R)injury to rat cardiomyocytes.
Methods
Cardiomyocytes isolated from the hearts of pathogen-free Sprague-Dawley rats(aged 1-3 days)were primarily cultured for 4 consecutive days and then divided into 6 groups(n=36 each)on the 5th day using a random number table: control group(group C), OGD/R group, OGD/R plus inactivated CORM(iCORM-3)50 μmol/L group(group iC), OGD/R plus CORM-3 12.5 μmol/L group(group C1), OGD/R plus CORM-3 25.0 μmol/L group(group C2), and OGD/R plus CORM-3 50.0 μmol/L group(group C3). The cardiomyocytes were incubated in glucose- and serum-free DMEM for 3 h and then in low-glucose DMEM for 6 h. At the beginning of oxygen-glucose deprivation, the corresponding concentrations of CORM-3 were added to the glucose-free DMEM in C1, C2 and C3 groups and iCORM-3(final concentration 50 μmol/L)were added to the glucose-free DMEM in group iC, and the glucose-free DMEM with the intervention drug was abandoned immediately after restoration of oxygen and glucose supply.At 6 h after restoration of oxygen and glucose supply, the pathological changes of cardiomyocytes were observed, cell proliferation was measured by CCK-8 assay, the activity of lactate dehydrogenase(LDH)in the supernatant was determined by lactic acid-pyruvate method, and the early apoptosis rate of cells was detected by flow cytometry.
Results
Compared with group C, the cell proliferation was significantly reduced, and the LDH concentration in the supernatant and early apoptosis rate were increased in the other five groups(P<0.05). Compared with OGD/R and iC groups, the cell proliferation was significantly increased, and the LDH concentration in the supernatant and early apoptosis rate were decreased in C1, C2 and C3 groups(P<0.05). The cell proliferation was significantly lower, and the LDH concentration in the supernatant and early apoptosis rate were higher in C2 and C3 groups than in group C1(P<0.05)and in group C3 than in group C2(P<0.05). The pathological changes of cardiomyocytes were significantly attenuated in C1, C2 and C3 groups, especially in group C1.
Conclusion
CORM-3 can reduce OGD/R injury to cardiomyocytes of rats, and the effect is gradually weakened with the increasing concentration.
Key words:
Carbon monoxide; Myocardial reperfusion injury
Publication Year: 2017
Publication Date: 2017-09-20
Language: en
Type: article
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