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Abstract

The Heat Shock Proteins (HSP, stress proteins) are considered as molecular chaperones because their complex function is essential to cell survival both in physiological and in stress conditions.

In order to evaluate a possible role of HSPs in the protection from ischemia-reperfusion injury, we developed an analytical procedure for their quantitative evaluation in human cardiac tissue.

The concentrations of Hsp 27 (related to cytoskeletal structures), Hsp 60 (expressed mainly at mythocondrial level), and both constitutive (Hsp 70) and inducibile (Hsp 72) forms of Hsp 70 family were measured in right atrium obtained from paediatric patients with congenital heart disease, submitted to open heart surgery, before and after cardioplegic arrest (CEC). To reduce inter-individual variability only patients with inter-atrial and inter-ventricular defects were admitted to the study. Cold St. Thomas cardioplegic solution was used in all cases. HSPs were measured by Western Blot analysis, after tissue extraction and protein separation by SDS-PAGE. Blots were developed by a colorimetric system, after an amplification step. After blot scanning, the NIH Image Program was used to analyze the blot. The quantitative evaluation of protein levels in the unknown samples was made by interpolation of a dose-response curve, performed in each blot using the respective standard protein. The following table reports the values (μg/mg protein, mean±SEM) of the measured HSPs:

SampleHsp 72Hsp 70Hsp 60Hsp 27
n = 17n = 17n = 17n = 20
Before CEC9.3 ± 1.53.7 ± 0.74.8 ± 0.82.4 ± 0.2
After CEC8.3 ± 1.64.5 ± 0.76.3 ± 1.12.7 ± 0.4
SampleHsp 72Hsp 70Hsp 60Hsp 27
n = 17n = 17n = 17n = 20
Before CEC9.3 ± 1.53.7 ± 0.74.8 ± 0.82.4 ± 0.2
After CEC8.3 ± 1.64.5 ± 0.76.3 ± 1.12.7 ± 0.4
SampleHsp 72Hsp 70Hsp 60Hsp 27
n = 17n = 17n = 17n = 20
Before CEC9.3 ± 1.53.7 ± 0.74.8 ± 0.82.4 ± 0.2
After CEC8.3 ± 1.64.5 ± 0.76.3 ± 1.12.7 ± 0.4
SampleHsp 72Hsp 70Hsp 60Hsp 27
n = 17n = 17n = 17n = 20
Before CEC9.3 ± 1.53.7 ± 0.74.8 ± 0.82.4 ± 0.2
After CEC8.3 ± 1.64.5 ± 0.76.3 ± 1.12.7 ± 0.4

Heat Shock Proteins, cardioplegia, ischemia-reperfusion injury
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© American Journal of Hypertension, Ltd. 2001
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