We recently reported that vanadium (V5+) was able to decrease the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-mediated induction of Cyp1a1 and Nqo1 at mRNA, protein and catalytic activity levels in mouse hepatoma Hepa 1c1c7 and human hepatoma HepG2 cells. However, little is known regarding the in vivo effects. Thus, the objective of this study was to investigate whether similar effects would occur at the in vivo level. Therefore, we examined the effect of exposure to V5+ (5 mg kg−1) with or without TCDD (15 µg kg−1) on the AhR-regulated genes in kidney, lung and heart of C57BL/6 J mice. Our results demonstrated that V5+ alone significantly decreased Cyp1b1 protein and catalytic activity levels in kidney at 24 h. Moreover, it significantly potentiated Nqo1 and Gsta1 gene expression in the heart, and only Gsta1 gene expression in the lung. Upon co-exposure, we found that V5+significantly inhibited the TCDD-mediated induction of Cyp1a1, Cyp1a2 and Cyp1b1 mRNA, protein and catalytic activity levels in the kidney at 24 h. On the other hand, V5+ significantly potentiated the TCDD-mediated induction of Nqo1 and Gsta1 protein and activity levels in the kidney. Cyp1a1, Cyp1b1, Nqo1 mRNA, protein and catalytic activity levels in the lung were significantly potentiated at 6 h. Interestingly, all tested genes in the heart were significantly decreased at 6 h with the exception of Gsta1 mRNA. The present study demonstrates that V5+ modulates TCDD-induced AhR-regulated genes. Furthermore, the effect on one of these enzymes could not be generalized to other enzymes even if it was in the same organ.

NAD(P)H:quinone oxidoreductase (NQO1)-mediated detoxification of quinones plays a critical role in cancer prevention. Heavy metals such as mercury (Hg(2+)) alter the carcinogenicity of aryl hydrocarbon receptor ligands, mainly by modifying various xenobiotic-metabolizing enzymes such as NQO1. Therefore, we examined the effect of Hg(2+) on the expression of NQO1 in human hepatoma HepG2 cells. For this purpose HepG2 cells were incubated with various concentrations of Hg(2+) (2.5, 5, and 10μM) in the presence and absence of two NQO1 inducers, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and isothiocyanate sulforaphane (SUL), as bifunctional and monofunctional inducers, respectively. Analysis of the time-dependent effect of Hg(2+) revealed that Hg(2+) increased the expression of NQO1 mRNA in a time-dependent manner. In addition, Hg(2+) increased NQO1 at the mRNA, protein, and activity levels in the presence and absence of both NQO1 inducers, TCDD and SUL, which coincided with increased nuclear accumulation of Nrf2 protein. Investigating the effect of Hg(2+) at the transcriptional level revealed that Hg(2+) significantly induced the antioxidant-responsive element-dependent luciferase reporter gene expression in the absence and the presence of both NQO1 inducers. NQO1 mRNA and protein decay experiments revealed a lack of posttranscriptional and posttranslational mechanisms. Transfecting HepG2 cells with siRNA for Nrf2 significantly decreased the Hg(2+)-mediated induction of NQO1 mRNA and catalytic activity by approximately 90%. In conclusion, we demonstrated that Hg(2+) regulates the expression of the NQO1 gene through a transcriptional mechanism in human hepatoma HepG2 cells. In addition, Nrf2 is involved in the modulation of NQO1 by Hg(2+).

Aryl hydrocarbon receptor (AhR) ligands such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and metals, such as mercury (Hg(2+)), are environmental co-contaminants and their molecular interaction may disrupt the coordinated regulation of the carcinogen-activating enzyme cytochrome P450 1A1 (CYP1A1). Therefore, we examined the effect of co-exposure to Hg(2+) and TCDD on the expression of the CYP1A1 in HepG2 cells. Our results showed that Hg(2+) significantly inhibited the TCDD-mediated induction of CYP1A1 at the mRNA, protein, and catalytic activity levels. At the transcriptional level, co-exposure to Hg(2+) and TCDD significantly decreased the TCDD-mediated induction of AhR-dependent luciferase reporter gene expression. Moreover, Hg(2+) did not affect CYP1A1 mRNA stability, while decreasing its protein half-life, suggesting the involvement of a posttranslational mechanism. Importantly, Hg(2+) increased the expression of heme oxygenase-1 (HO-1), a rate limiting enzyme in heme degradation, which coincided with further decrease in the CYP1A1 catalytic activity levels. Upon using a competitive HO-1 inhibitor, tin mesoporphyrin, heme precursor, hemin, or transfecting the HepG2 cells with siRNA for HO-1 there was a partial restoration of the inhibition of TCDD-mediated induction of CYP1A1 catalytic activity. In conclusion, we demonstrate that Hg(2+) down-regulates the expression of CYP1A1 at the transcriptional and posttranslational levels in HepG2 cells. In addition, HO-1 is involved in the modulation of CYP1A1 at the posttranslational level.

Objective: The present study investigates the effects of alprazolam (ALP) and nifedipine alone or in combination on behavior and on g-aminobutyric acid (GABA) levels, in discrete brain regions of albino rats.

Methods: The anxiolytic effect was studied using a plus maze model and brain levels of GABA were measured using high performance liquid chromatography. Four acute treatment groups of rats were used. In the first they were treated with 1% Tween 80 (1ml/kg), in the second with nifedipine (10mg/kg), in the third with ALP (2mg/kg) and in the fourth with ALP in addition to nifedipine in the respective doses. The work was carried out at the Faculty of Pharmacy of Al-Fateh University, Tripoli, Libya in the first half of 2002.

Results: The results indicate that the anxiolytic effect of ALP was not modified by nifedipine. Nifedipine by itself significantly decreased the motor activity (decrease in total lines crossed), this effect was apparently antagonized by ALP. Alprazolam administration produced an increase of GABA levels in cerebellum and striatum and a decrease in the brain stem. Nifedipine per-se had no effect on GABA levels in the brain stem but it partially antagonized ALP-induced inhibitory effect on GABA in this region. Alprazolam significantly increased GABA levels in the striatum, while nifedipine alone had no effect on neurotransmitter levels and did not modify the ALP effect in this brain region. Alprazolam or nifedipine had no significant effect on GABA levels in midbrain, cerebral cortex and whole brain. There were no significant changes in GABA levels in midbrain and whole brain with drug combination. However, the combination decreased GABA levels significantly in the cerebral cortex.

Conclusion: It may be concluded that, the anxiolytic effect of ALP possibly occurs through changes in brain GABA levels (an increase in cerebellum and striatum with a decrease in brain stem). The effect was not modified by nifedipine which per se had no affect on GABA levels in any brain area. The significant decrease in GABA levels in cerebral cortex by ALP-nifedipine combination may be due to the mutual closure of calcium channel (mentioned in literature) resulting in inhibition of the EAA-ergic input to GABA-ergic neuron.