Quantifying Cardiomyocyte Apoptosis in Ischemic Heart

Cardiomyocyte apoptosis in the area at risk (AAR) has a significant effect on myocardial survival and function. After 3 h of reperfusion, tissue samples from the area at risk (AAR) were analyzed. Cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl nick-end labeling (TUNEL) and caspase-3 activity assay. TUNEL labeling was performed as described in previous study [6 (link), 20 (link)] by using an in situ cell death detection kit (Roche). In brief, the slides were incubated with TUNEL reaction mixture and then counterstained with the 4′,6-diamino-2-phenylindole (DAPI) to detect the nuclei. The apoptosis index was calculated as a percentage of the number of TUNEL-positive apoptotic cells over the total number of nucleated cells (DAPI staining). Myocardial caspase-3 activity was determined as described before [21 (link)] by using a caspase colorimetric assay kit (Chemicon, Temecula, CA, USA) according to manufacturer’s protocol.

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Ding M., Lei J., Han H., Li W., Qu Y., Fu E., Fu F, & Wang X. (2015). SIRT1 protects against myocardial ischemia–reperfusion injury via activating eNOS in diabetic rats. Cardiovascular Diabetology, 14, 143.

Publication 2015

in situ cell death detection kit caspase colorimetric assay kit

Apoptotic Assay Cardiomyocyte Caspase Caspase 3 Cell death Colorimetric Myocardial Nuclei Reperfusion Tissue Tunel

Corresponding Organization :

Other organizations : Air Force Medical University, Xijing Hospital, Xian Central Hospital, First Hospital of Xi'an

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Cardiomyocyte apoptosis in the area at risk (AAR) detected by terminal deoxynucleotidyl nick-end labeling (TUNEL)
Caspase-3 activity in the myocardium

control variables

None explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

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