Risk of overestimation of free malondialdehyde in perfused rat hearts due to homogenization artifacts.

Autor:	Verbunt RJ; Department of Cardiology, University Hospital Leiden, Netherlands., Egas JM, Van der Laarse A
Jazyk:	angličtina
Zdroj:	Cardiovascular research [Cardiovasc Res] 1996 Apr; Vol. 31 (4), pp. 603-6.
Abstrakt:	Objectives: The aims of the present study were to determine (1) whether free malondialdehyde (MDA) was artifactually formed during homogenization of myocardial tissue and (2) whether free MDA was increased in reperfused rat hearts. Methods: Groups of isolated buffer-perfused rat hearts were subjected to control perfusion, or 20 min of ischemia, or 20 min of ischemia followed by 5 or 30 min of reperfusion. The hearts were subsequently assayed for free MDA by ion-pairing high-performance liquid chromatography following homogenization in the absence or presence of the antioxidant butylated hydroxytoluene (0.01%). Results: Tissue homogenates prepared in the absence of butylated hydroxytoluene contained significantly higher (P < 0.001) free MDA levels than tissue homogenates from the same hearts prepared in the presence of butylated hydroxytoluene. Free MDA levels of tissue homogenates prepared in the presence of butylated hydroxytoluene were below the detection limit (20 pmol/mg protein) in 27 of 30 tissue homogenates, irrespective of the perfusion protocol. Control experiments showed that the presence of butylated hydroxytoluene did not interfere with the detection of free MDA. Conclusions: These results indicate that free MDA was formed artifactually during tissue homogenization in the absence of butylated hydroxytoluene. Furthermore, free MDA could not be detected in perfused rat hearts after control perfusion, or 20 min of ischemia, or 20 min of ischemia followed by 5 or 30 min of reperfusion.
Databáze:	MEDLINE
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