Curcumin (diferuoylmethane) is a natural compound with anticareinogenic activities which is able to exert either proapoptotic or antiapoptotic effects in different cell types. This paper focuses on the sequence and extent of primary events induced by curcumin, in comparison with those occurring during dexamethasone-induced apoptosis in rat thymocytes. It also presents annexin VI-FITC as a new probe for studying membrane asymmetry. Curcumin readily penetrates into the cytoplasm, and is able to accumulate in membranous structures such as plasma membrane, endoplasmic reticulum and nuclear envelope. Curcumin-treated cells exhibit typical features of apoptotic cell death, including shrinkage, transient phosphatidylserine exposure, increased membrane permeability and decrease in mitochondrial membrane potential. However, nuclei morphology, DNA fragmentation, the extent and time-course of membrane changes are different from those observed during dexamethasone-induced apoptosis, suggesting that, despite many similarities, the mode of action and the events triggered by cnrcumin are different from those occurring during typical apoptosis.© In this paper, we analyzed the primary effects of the presence of CUR in rat thymocytes, and compared the sequence and extent of CUR-evoked events with those occurring during the typical apoptotic program induced by dexamethasone (DEX). The fluorescence and lipophilic properties of CUR [12] provided the basis for the search of its cellular localization and for analyzing the local effects on the structure and function of cellular membranes. Plasma membrane asymmetry was studied by a new fluorescence probe, i.e. annexin VI (Anx VI), isolated from porcine liver and coupled with FITC. We could investigate other parameters such as plasma membrane permeability, mitochondrial membrane potential, as well as other typical apoptotic features ,i.e. changes in cell volume, nucleus morphology and DNA fragmentation.
Materials and methods