Effects of antioxidants and MAPK inhibitors on cell death and reactive oxygen species levels in H(2)O(2)-treated human pulmonary fibroblasts

H(2)O(2)-induced cytotoxicity in normal human pulmonary fibroblasts (HPFs) is of interest in toxicological research since HPFs are involved in lung inflammation, fibrosis and cancer. The present study investigated the cytotoxic effects of H(2)O(2) on normal HPFs in relation to reactive oxygen species (ROS) and mitogen-activated protein kinases (MAPKs) using the well-known antioxidants N-acetyl cysteine (NAC) and propyl gallate (PG), as well as MAPK inhibitors. Treatment with 50 μM H(2)O(2) inhibited the growth of the HPFs by ∼45% in 24 h. H(2)O(2) induced cell death via apoptosis and triggered the loss of mitochondrial membrane potential (MMP; Δψ(m)) in the HPFs. H(2)O(2) also increased the ROS levels, including O(2)(•−), in the HPFs and induced glutathione (GSH) depletion. NAC and PG attenuated the death of the HPFs and the loss of MMP (Δψ(m)) through the use of H(2)O(2). NAC decreased the ROS levels in the H(2)O(2)-treated HPFs and PG markedly prevented an increase in O(2)(•−) levels in these cells. However, PG alone induced cell death in the HPF control cells and increased the ROS levels in these cells. None of the MAPK (MEK, JNK and p38) inhibitors affected cell growth inhibition or cell death by H(2)O(2). In addition, these inhibitors did not significantly affect the ROS levels and GSH depletion in the H(2)O(2)-treated HPFs. In conclusion, H(2)O(2) induced growth inhibition and cell death in the HPFs via GSH depletion. NAC and PG attenuated H(2)O(2)-induced HPF cell death but each regulated the ROS levels in a different manner. Treatment with MAPK inhibitors did not affect cell death or the ROS levels in the H(2)O(2)-treated HPFs.