Figure 3

Inhibition of thioredoxin reductase by DNCB and pretreatment of cells with DHA rescues asbestos-induced oxidation of Trx1. (A) LDH assay to assess lytic cell death after pretreatment of LP9 cells with DNCB and exposure to asbestos (data is presented as a percentage of the lytic control). (B) Effect of DNCB on asbestos-induced Trx1 oxidation. Cells were pretreated with 10 μM DNCB for an hour and then exposed to asbestos for 8 h. Cell lysates were then derivatized with IAA and analyzed for oxidation of Trx1 by redox Western blot and densitometry of redox Western analysis of Trx1 oxidation state was performed. (C) Effect of DNCB pretreatment on chrysotile asbestos-induced oxidation of Trx1 (D) Analysis of apoptosis in response to DNCB pretreatment and asbestos exposure for 8 h as measured by Apostain technique. (E) LP9 cells were pretreated with 1 mM DHA for an hour and exposed to asbestos for 8 h. Thereafter, the oxidation state of Trx1 was assessed by redox Western blot analysis (*p < 0.05 compared to null controls; †p < 0.05 compared to crocidolite asbestos exposure alone (Croc 75 n = 2 per group).