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TEGDMA Modulates Glutathione Transferase P1 Activity in Gingival Fibroblasts

¹ Faculté de Chirurgie Dentaire, Laboratoire de Biologie et Physiopathologie Cranio-Faciale, 1 rue Maurice Arnoux, F-92120 Montrouge, France;
² Centre Universitaire des Saints-Pères, INSERM U 490, Toxicologie Moléculaire et Service de Biochimie, Hôpital Européen Georges Pompidou, Paris, France; and
³ Institut Cochin, CNRS UMR 8104, INSERM U 567, Paris, France;

View larger version (22K): [in a new window]   Figure 1. Effect of TEGDMA on GSH and GSSG levels in fibroblasts and in a cell-free system. (Panel A) Time-course of GSH and GSSG content in gingival fibroblasts treated with TEGDMA at a TD50 concentration (0.3 mM). (Panel B) Time-course of depletion of GSH induced by 3 mM TEGDMA at 37°C in a cell-free system. The decrease of GSH at 1 and 3 hrs was significant (P < 0.0001 and P < 0.00001, respectively). (Panel C) Depletion of GSH induced by various concentrations of H2O2. (Panel D) The effect of TEGDMA on GSH in the presence or absence of GST P1. *The difference between TEGDMA+GST and TEGDMA alone was statistically significant (P < 0.0001). Error bars are standard deviation (for each group, n = 3). Data are representative of 3 separate experiments.

View larger version (15K): [in a new window]   Figure 2. Modulation of the activity of fibroblast GSTP1 variants by TEGDMA. The cells were treated with various TEGDMA concentrations or DMSO (control) for 90 min. Data represent the means (± SD) of triplicate experiments per condition and are expressed as % of the respective control values (*A/*A, 137.05 ± 11.9; *A/*B, 112.2 ± 5.2; and *B/*B, 110.7 ± 19.4 nmoles/min/mg). *P < 0.05; **P < 0.01.

View larger version (23K): [in a new window]   Figure 3. Effect of TEGDMA on GSTP1 activity in a cell-free system. (Panel A) Time-course activity of purified GSTP1 after TEGDMA treatment at the concentration of 1 mM. (Panel B) GSTP1 activity following treatment with various concentrations of TEGDMA for 90 min. (Panels C,D) TEGDMA-induced inhibition of GST activity as a function of concentrations of GSH and CDNB, respectively. Inserts in panels C and D show the Lineweaver-Burk representation of the data. The data represent the means (± SD) of triplicate experiments.

View larger version (11K): [in a new window]   Figure 4. Effect of TEGDMA on the viability of fibroblasts. The viability of gingival fibroblasts treated with various concentrations of TEGDMA for 24 hrs was determined by the MTT assay. Each result represents the TEGDMA TD50 value obtained with cells expressing GSTP1 *A/*A (n = 9), *A/*B (n = 9), and *B/*B (n = 5). *Indicates a significant difference between the *B/*B and *A/*A groups (P < 0.05). The mean values are indicated by dashes.