Figure 3.

N,N′-bis(2-Mercaptoethyl)isopthalamide (NBMI) attenuates the mercury-induced phospholipase D (PLD) activation in similar fashion to N-acetyl-l-cysteine (NAC) and meso-2,3-dimercaptosuccinic acid (DMSA) in mouse aortic endothelial cells (MAECs). The MAECs (5 × 10⁵ cells/35 mm dish) were labeled with [³²P]orthophosphate in phosphate-free Dulbecco-modified Eagle medium (DMEM) for 12 hours. Following [³²P]orthophosphate labeling, cells were pretreated with minimal essential medium (MEM) alone or MEM containing NBMI (50 μmol/L) and MEM containing NAC (50 μmol/L, A and 5 mmol/L, B) or DMSA (50 (C) μmol/L and 5 mmol/L (D)) for 1 hour. After pretreatment, cells were treated with MEM alone or MEM containing methylmercury (10 μmol/L) or thimerosal (25 μmol/L) for 1 hour in the presence of 0.05% (volume/volume [vol/vol]) 1-butanol. At the end of the incubation period, [³²P]phosphatidylbutanol ([³²P]PBt) formed was determined. Data represent mean ± standard deviation (SD) calculated from 3 independent experiments. *Significantly different at P < .05 as compared to cells treated with MEM alone. **Significantly different at P < .05 as compared to cells treated with MEM containing mercury alone.