Effect of Aminoethylcysteine Ketimine Decarboxylated Dimer, a Natural Sulfur Compound Present in Human Plasma, on Tert-Butyl Hydroperoxide-induced Oxidative Stress in Human Monocytic U937 Cells

Abstract

Aminoethylcysteine ketimine decarboxylated dimer (AECK-DD) is a natural sulphur compound present in human plasma and urine and in mammalian brain. Recently, it has been detected in many common dietary vegetables. The aim of the present study was to evaluate the ability of AECK-DD to affect cellular response of U937 human monocytic cells to tert-butyl hydroperoxide-induced oxidative stress. AECK-DD was incorporated into cells, as confirmed by GC-MS analyses, without any cytotoxic effect. A 24 h treatment with 50 and 250 μM AECK-DD resulted in the incorporation of 0.10±0.01 and 0.47±0.08 ng AECK-DD × 10⁶ cells, respectively. U937 cells pretreated with AECK-DD (in the range 4–100 μM) showed an increased resistance to tert-butyl hydroperoxide-induced necrotic death, as revealed by a higher percent of survival measured at all incubation times with respect to control cells. Moreover, the protective effect exhibited by AECK-DD is significantly stronger with respect to that obtained with other common antioxidants (N-acetyl cysteine and trolox) and comparable, although somewhat higher, to that of vitamin E. This effect seems to be due to the ability of AECK-DD to reduce glutathione depletion and to inhibit lipid peroxidation during tert-butyl hydroperoxide treatment. It can be concluded that AECK-DD protects cultured human monocytic cells against tert-butyl hydroperoxide-induced oxidative stress and subsequent cell death, likely through an antioxidant action inside the cell. Due to its presence in both human plasma and urine, AECK-DD may play a role in the modulation of oxidative processes in vivo.

• Aminoethylcysteine ketimine decarboxylated dimer
• U937
• tert-Butyl hydroperoxide
• Oxidative stress
• Natural antioxidants
• AECK-DD, aminoethylcysteine ketimine decarboxylated dimer
• t-BOOH, tert-butyl hydroperoxide
• PBS, Dulbecco's phosphate-buffered saline
• NAC, N-acetyl cysteine
• GSH, reduced glutathione
• GSSG, oxidized glutathione
• GPX, glutathione peroxidase
• GR, glutathione reductase
• CAT, catalase
• CumOOH, cumene hydroperoxide
• HPLC-ECD, high performance liquid chromatography-electrochemical detector
• GC-MS, gas chromatography-mass spectrometry
• LDH, lactate dehydrogenase
• NADH, β-nicotinamide adenine dinucleotide, reduced form
• TBA-RS, thiobarbituric acid-reactive substances
• DETAPAC, diethylenetriaminepentaacetic acid
• DCF, 2′7′-dichlorofluorescein